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Issue 22 ， 2019
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Effect of Modified Chaihu Shugantang Regulat miRNA-204 on Hippocampus Autophagy in Epileptic Mice
Abstract：Objective:To observe the effect of modified Chaihu Shugantang on the expression of miRNA-204 in hippocampus of epileptic mice, and to explore its mechanism of neuroprotection.Method:The sixty mice were randomly divided into 6 groups: normal group, model group (pilocarpine 180 mg·kg-1), and modified Chaihu Shugantang group (7 g·kg-1·d-1), modified Chaihu Shugantang+ miRNA-204 mimic group (7 g·kg-1·d-1+ 2 μL), modified Chaihu Shugantang+ miRNA-204 inhibitor group (7 g·kg-1·d-1+ 2 μL), carbamazepine group (30 mg·kg-1·d-1), each was given intragastric administration for 2 weeks, using pilocarpine to cause epilepsy in mice, respectively, add flavor to Bupleurum after intragastric administration, inhibition and overexpression of miRNA-204, the mice were sacrificed and their hippocampus tissues were harvested.The indicators of each group were observed, Real-time quantitative PCR detecting system (Real-time PCR) was used to detect mouse hippocampal miRNA-204 expression, Western blot analysis of autophagy-related protein microtubule-associated protein light chain 3 (LC3), autophagy-associated marker protein 7 (ATG7) expression, hematoxylin pathological condition of hippocampus in each group was observed by hematoxylin-eosin(HE)staining.The autophagy of hippocampus in each group was observed by transmission electron microscopy.Result:Compared with normal group, the expression of miRNA-204 was significantly decreased in model group (P<0.01), the pathological changes in the hippocampal C1 area were the most obvious, the expression of ATG7, LC3Ⅱ/LC3Ⅰ was increased (P<0.01), and the autophagy was small. Compared with model group, the expression of miRNA-204 in the hippocampus of the modified Chaihu Shugantang group was increased (P<0.05), the pathological changes in the hippocampal C1 area were alleviated, the expression of ATG7, LC3Ⅱ/LC3Ⅰ was decreased (P<0.05), and the autophagy was small. The number of body decreased, the expression of miRNA-204 in hippocampus of modified Chaihu Shugantang+ miRNA-204 mimic group was significantly increased (P<0.01), the pathological changes in hippocampal C1 area were the lightest, and the expression of ATG7, LC3II/LC3I was decreased (P<0.01), the number of autophagosomes was the least.Compared with modified Chaihu Shugantang group, the above-mentioned indicators of modified Chaihu Shugantang+ miRNA-204 inhibitor group had the same change trend and the change range decreased (P<0.05).Conclusion:Modified Chaihu Shugantang can improve the pathological changes of hippocampus in mice with epilepsy and play a neuroprotective role. The mechanism may be to increase the expression of miRNA-204 in hippocampus of mice with epilepsy, inhibit excessive autophagy of neurons and reduce apoptosis.Keywords：Chaihu Shugantang;epilepsy;miRNA-204;autophagy- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Hei Xiaoyaosan on Expressions of APP, PERK in Hippocampus Area of Alzheimer's Dementia Mice
Abstract：Objective:To study the effect of Hei Xiaoyaosan on endoplasmic reticulum stress in hippocampal neurons of Alzheimer's disease (AD) model mice, including behavioral, histopathology and amyloid precursor protein (APP), protein kinase endoplasmic reticulum kinase (PERK) expressions.Method:The 42 4-month-old SPF-grade double transgenic (APP/PS1) mice were randomly divided into the high-dose group and the low-dose group, the donepezil hydrochloride group and the model group, and 10 C57BL mice of the same age were used as the blank group. Firstly, they were adapted to the environment for one week. After 2 months of treatment with different drug interventions, Morris water maze behavior was used to test the learning and memory abilities of each group of mice. After 1 month of treatment, histopathological changes in the hippocampus of each group of mice were observed by light microscopy. The expressions of APP, PERK protein and mRNA in the endoplasmic reticulum of hippocampus were detected by immunohistochemistry (IHC) and real-time fluorescent quantitative polymerase chain reaction (Real-time PCR).Result:After drug intervention, compared with the blank group, the escape latency of the AD model group was significantly prolonged (P<0.01), the neurons in the hippocampus of the model group were severely damaged, and the expressions of APP and PERK in the model group were significantly increased (P<0.01). In the comparison model group, the escape latency of the mice in each treatment group was significantly shortened (P<0.05, P<0.01), the degree of neuronal damage in the hippocampus of each treatment group was alleviated, and the positive expressions of APP and PERK in each treatment group were weakened (P<0.05).Conclusion:Hei Xiaoyaosan can significantly improve the learning and memory abilities of AD mice, which may be related to the reduction of the excessive stress response of endoplasmic reticulum to alleviate cognitive impairment in AD mice.Keywords：Alzheimer's disease;Hei Xiaoyaosan;endoplasmic reticulum stress- 5
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Pinelliae Rhizoma Praeparatum Cum Alumine Polysaccharides on MUC5AC mRNA in Lung Tissues of Allergic Asthma Model Rats
Abstract：Objective:To observe the effect of Pinelliae Rhizoma Praeparatum Cum Alumine polysaccharides(PRPCAP)on airway mucus secretion in rats with allergic asthma, in order to study the material basis of the " macromolecule" component of the polysaccharides as the original medicinal materials.Method:The 60 SPF-grade Wistar rats were induced by intraperitoneal injection of chicken ovalbumin (OVA) and aluminum-magnesium adjuvant, except for the normal control group. The OVA solution was aerosolized to establish a rat model of allergic asthma. After successful modeling, the rats were randomly divided into 5 groups, namely allergic asthma model group, positive drug group (montalurast sodium, 5 mg·kg-1), high-dose PRPCAP group (400 mg·kg-1), middle-dose PRPCAP group (200 mg·kg-1) and low-dose PRPCAP group (100 mg·kg-1). The contents of interleukin-4 (IL-4) and interferon-γ (IFN-γ) in serum and bronchoalveolar lavage fluid (BALF) supernatant were determined by enzyme-linked immunosorbent assay (ELISA), and the count of eosinophils (EOS) was detected by BALF sediment. The histopathological changes were observed by hematoxylin-eosin (HE) staining in lung tissue. The mRNA expression of mucin 5AC (MUC5AC) was detected by Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR).Result:Compared with the normal control group, serum IL-4 level in the allergic asthma model group was significantly increased (P<0.01), IFN-γ level was significantly decreased (P<0.01), and EOS counts in BALF were significantly increased (P<0.01). Compared with the allergic asthma model group, the positive drug group, and high, middle, and low-dose PRPCAP groups could significantly reduce the serum IL-4 content (P<0.01), while high and middle-dose PRPCAP groups could significantly reduce the content of IL-4 in BALF (P<0.05, P<0.01), the middle-dose PRPCAP group could significantly increase the content of IFN-γ in serum (P<0.05), EOS counts in BALF were significantly lower in the positive drug group and all of PRPCAP groups (P<0.01). HE staining showed the pathological changes in the lung tissue of the allergic asthma model group and a mitigating effect in the positive drug group and all of PRPCAP groups. Compared with the normal control group, the expression of MUC5AC mRNA in the lung tissue of rats with allergic asthma was significantly increased (P<0.01). Compared with the model group of allergic asthma rats, the positive drug group and all of PRPCAP groups showed significant decrease in MUC5AC mRNA (P<0.01).Conclusion:The " macromolecule" component of polysaccharides in the Pinelliae Rhizoma Praeparatum Cum Alumine may be the material basis for the efficacy of eliminating dampness and eliminating phlegm.Keywords：Pinelliae Rhizoma Praeparatum Cum Alumine polysaccharides;allergic asthma model rat;mucin 5AC (MUC5AC)- 12
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Tangnaikang on Imbalance Between NE and α1-AT in Spontaneous Type 2 Diabetic Mouse
Abstract：Objective:To observe the regulatory effect of Tangnaikang (TNK) on imbalance between neutrophil elastase (NE) and α1-antitrypsin (α1-AT) in ob/ob mice with type 2 diabetes mellitus (T2DM).Method:Thirty-two male SPF ob/ob mice were randomly divided into model group (DM, normal saline) and high-dose TNK group (TNKH, TNK solution 16.04 g·kg-1), middle-dose TNK group (TNKM, TNK solution 8.02 g·kg-1) and low-dose TNK group (TNKL, TNK solution 4.01 g·kg-1). Another 8 C57BL/6J mice were included in normal group (Con, saline). The experiment lasted for four weeks. The general state, body weight (BW) and fasting blood glucose (FBG) of the mice were recorded weekly, the oral glucose tolerance (OGTT) test was performed on the 25th day, the insulin tolerance (ITT) test was performed on the 27th day, and the area under the curve (AUC) was calculated. After the end of the experiment, serum was used to detect the level of fasting insulin (Fins), insulin resistance index (HOMA-IR), total triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), NE and α1-AT. Adipose tissue was used to detect the expressions of NE, α1-AT, phosphor-insulin receptor substrate 1 antibody (p-IRS1) and glucose transporter 4 (GLUT4) proteins.Result:Compared with the Con group, the BW of the ob/ob mice of the model group increased significantly, the glucose and lipid metabolism indexes showed diabetes, the serum and adipose tissue NE increased significantly (P<0.01), and the α1-AT decreased significantly (P<0.01). Compared with the DM group, the BW, TG, TC, and LDL indexes in the TNKH group were significantly decreased (P<0.01), the HDL was significantly increased (P<0.01), and the FBG, Fins, HOMA-IR, AUCogtt and AUCITT were significantly decreased (P<0.01), serum and adipose tissue NE decreased significantly (P<0.01), α1-AT increased significantly (P<0.01), and adipose tissue p-IRS1 and GLUT4 increased significantly (P<0.05).Conclusion:TNK can reduce the BW of ob/ob mice, improve glycolipid metabolism, increase α1-AT level, decrease NE level, and regulate IRS1-GLUT4 signaling pathway, which may be one of its mechanisms in improving IR of adipose tissue mediated by neutrophil.Keywords：type 2 diabetes;Tangnaikang;ob/ob mice;neutrophil elastase;α1-antitrypsin- 5
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Drug-containing Serum of Clerodendrum bungei on Liver Cancer Cells Through PI3K/Akt Signaling Pathway
Abstract：Objective:To investigate the effect of Clerodendrum bungei-containing serum on liver cancer MHCC97-H cells and its possible mechanism from the perspective of phosphatidylinositol 3-kinase(PI3K)/protein kinase (Akt) signaling pathway.Method:The medicinal serum of 15% C. bungei was used to treat MHCC97-H cells. The effect of serum containing C. bungei on cell proliferation was observed by cell counting kit-8(CCK-8) method, in order to select the best time and concentration. The apoptosis was detected by Annexin V-FITC/PI double staining method. Western blot was used to detect the posphatase and tensin homologous gene deleted from chromosome 10 in key proteins (PTEN), phosphoprotein kinase B (p-Akt) and phosphatidylinositol 3-kinase (PI3K)-related protein expression of PI3K/Akt signaling pathway. Real-time PCR was used to detect C. bungei-containing serum on cells for 72 h after activation of nuclear factor-activated B cell kappa light chain(NF-κB) and tumor necrosis factor-α (TNF-α) mRNA expression.Result:The results of CCK-8 showed an inhibitory effect of the C. bungei-containing serum on the proliferation of tumor cells in a dose and time-dependent manner. Among them, the high-dose group had the most obvious inhibitory effect, and the maximum inhibition rates at 24, 48, 72 h were 28%, 32%, and 43%, respectively. The results of flow cytometry showed that with the increase of drug-containing serum concentration, the cell growth was observed. The inhibition rate of cells was increased to different degrees, and the inhibition effect was significantly increased in the 72 h intervention group (P<0.01). The maximum apoptosis rate of the C. bungei-containing serum group was 19.48% and 19.72%, compared with the blank group. The difference was significant (P<0.05). The results of Western blot showed that the expression of PTEN in different groups increased with different concentrations of C. bungei-containing serum (P<0.05), and the expressions of PI3K and p-Akt decreased (P<0.05). Real-time PCR results showed that the C. bungei-containing serum could down-regulate the expression of NF-κB and up-regulate the expression of TNF-α mRNA (P<0.05).Conclusion:The medicinal serum of C. bungei can effectively inhibit the proliferation of MHCC97-H hepatoma cells and promote its apoptosis, which may be related to the PI3K/Akt signaling pathway and its key factors.Keywords：Clerodendron bungei;liver cancer cell MHCC97-H;phosphatidylinositol 3-kinase(PI3K)/protein kinase (Akt) signaling pathway;drug-containing serum- 6
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Mechanism of Dichroa Alkali Salt Inducing Vomiting Based on Pica in Mice
Abstract：Objective:To study the possible mechanism of dichroa alkali salt (DAS) in inducing vomiting.Method:Mice pica model was used to observe the antagonistic effect of the three different kinds of antiemetic drugs [dopamine receptor antagonist metoclopramide, 5-hydroxytryptamine 3 (5-HT3) receptor antagonist ondansetron and neurokinin-1 receptor antagonist aprepitant] on body mass, food intake, kaolin consumption, diarrhea and death induced by DAS to preliminarily clarify the possible pathogenic pathway of DAS. Then, the expression of 5-HT and substance P(SP) in ileum and medulla of mice induced by DAS alone at different time points was detected by enzyme-linked immunosorbent assay to confirm whether DAS could affect the changes of these two neurotransmitters.Result:After treatment with ondansetron and aprepitant, DAS-induced reduction in food intake of mice was significantly improved on the 4th day after continuous administration and on the 1st day after drug administration (P<0.01), since the 3rd day after administration, DAS-induced body mass loss of mice was significantly improved (P<0.01), and DAS-induced diarrhea and mortality in mice were significantly reduced, while DAS-induced pica in mice was effectively antagonized. However, metoclopramide did not significantly improve the above indicators. Further detection of vomiting-related neurotransmitters found that compared with the blank group, the expressions of 5-HT at 4 h, 12 h and SP at 48 h after treatment with DAS were significantly increased in ileum and medulla of mice.Conclusion:The mice pica model can be used to effectively characterize DAS-induced vomiting. DAS-induced pica in mice may be associated with the increase of 5-HT and SP in ileum and medulla. Ondansetron and aprepitant can effectively antagonize DAS-induced pica in mice.Keywords：dichroa alkali salt;vomiting;pica in mice;compatibility;antiemetic drug- 5
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Mechanism of Trichosanthis Pericarpium Aqueous Extract in Protecting H9c2 Cardiomyocytes from Hypoxia/Reoxygenation Injury via PI3K/Akt/NO Signaling Pathway
Abstract：Objective:To investigate the effect of Trichosanthis Pericarpium aqueous extract(TPAE)in protecting H9c2 cells from hypoxia/reoxygenation (H/R) injury by activating phosphatidylionsitol-3-kinase/protein kinase B/nitric oxide(PI3K/Akt/NO) signaling pathway.Method:The 2.5 mmol·L-1 Na2S2O4 was used to induce the model of H9c2 cardiomyocytes H/R injury in the experiments. The cultured H9c2 cardiomyocytes were randomly divided into normal group, H/R group (model group) and inhibition group (LY294002, 10 μmol·L-1). In the H/R + TPAE group, 50 mg·L-1 TPAE was added to the cultures at 24 h before H/R exposure. Cell viability was measured by methyl thiazolyl tetrazolium (MTT) assay. The amounts of NO, endothelial nitric oxide synthase (eNOS), and induced nitric oxide synthase (iNOS) were tested by enzyme linked immunosorbent assay (ELISA) kits. Reverse transcription-quantitative real-time polymerase chain reaction (Real-time PCR) was performed to analyze relative mRNA expressions of Akt, eNOS and iNOS. Western blot was used to detect the expressions of Akt, p-Akt (Ser 473), eNOS, and iNOS.Result:Compared with the normal control group, the cell viability significantly decreased in the model control group (P<0.01), the release of NO was obviously down-regulated (P<0.01), the mRNA and protein expressions of p-Akt, Akt, eNOS were remarkably decreased (P<0.01), while those of iNOS were up-regulated (P<0.01). Compared with the H/R group, the pretreatment with TPAE remarkably improved the morphological lesion of cardiomyocytes, enhanced cell viability(P<0.01), increased the expressions of Akt, p-Akt (Ser 473) and eNOS(P<0.01), decreased the expression of iNOS(P<0.01), and increased the release of NO(P<0.01). The PI3K inhibitor LY294002 was used. Obviously, cardioprotection of Trichosanthis Pericarpium aqueous extract was blocked by co-treatment LY294002, and cell viability was correspondingly reversed.Conclusion:TPAE can protect H9c2 cardiomyocytes from H/R injury by activating PI3K/Akt signaling pathway, which might be related to the up-regulation of the mRNA and protein expressions of eNOS, the down-regulation of the level of iNOS, and the increase of the production of physiological amounts of NO.Keywords：Trichosanthis Pericarpium aqueous extract;hypoxia/reoxygenation injury;cardiomyocytes;phosphatidylionsitol-3-kinase/protein kinase B/nitric oxide(PI3K/Akt/NO)- 13
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Evaluation of Rat Model with Spleen Deficiency and Dampness Stagnancy Based on Bioelectrical Impedance
Abstract：Objective:To evaluate the model with spleen deficiency and dampness stagnancy by bioelectrical impedance analysis (BIA) and traditional indicators.Method:The forty rats were divided into blank group and model group, with 20 rats in each group. The rats in the blank group were fed with normal feed, the rats in model group were prepared with the spleen deficiency and dampness stagnancy model for 14 days. Observe the general condition of the rats, measure the water content of the feces in the dry method, measure the water load index by weighing method, and detect the urinary D-xylose excretion total protein (TP), albumin (Alb) content, by enzyme-linked immunosorbent assay (ELISA). Western blot analysis of renal aquaporin 1 (AQP1) content, and the use of experimental animal body composition analyzer to determine the total water content (TBW), extracellular fluid (ECF), intracellular fluid (ICF), fat mass (FM), free fat mass (FFM) and body mass bioelectrical impedance index such as body mass index (BMI).Result:Compared with blank group, the rats in model group lost weight, gradually loose stools occasionally, the anus temperature was basically unchanged, body mass, D-xylose excretion, water load index, TP and Alb content decreased (P<0.01). The feces contained water, the rate and the content of AQP1 in the renal pulp were increased (P<0.01). The TBW, ECF and FFM were increased, and the ICF, FM and BMI were decreased (P<0.01).Conclusion:Rats with spleen deficiency and dampness stagnancy induced a combination of factors such as diet and excessive fatigue. The bioelectrical impedance method can be more intuitive and comprehensive.Keywords：spleen deficiency and dampness stagnancy;bioelectrical impedance;experimental animal body composition determination analyzer;water liquid metabolism- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Pharmacology
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Effect of Wenfei Zhiliudan in Treating Lung Qi Deficiency Cold Type Allergic Rhinitis and Its Mechanism
Abstract：Objective:To observe the efficacy and safety of Wenfei Zhiliudan in treatment of lung Qi deficiency cold type allergic rhinitis.Method:A total of 120 cases were randomly divided into control group and observation group, with 60 cases in each group. The control group was given desloratadine + momethasone furoate, while observation group was given Wenfei Zhiliudan. A course of treatment was 2 weeks. The rhino conjunctivitis quality of life questionnaire (RQLQ), total nasal symptom score (TNSS) and traditional Chinese medicine (TCM) syndrome differentiation scale for lung Qi deficiency cold (TCM scale) in two groups before and after treatment were observed. The serum inflammatory factor [tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), interleukin-33 (IL-33)] and immune indexes [immunoglobulin E (IgE), eosinophils (EOS), eotaxin (EOT)] were determined. The total effective rate and the incidence of adverse reactions were compared between two groups.Result:Five cases felt off during the study period. The total effective rate in observation group was 91.5%(54/59), which was higher than 82.5%(47/57) in control group (P<0.05). The RQLQ, TNSS and TCM scale scores in observation group were significantly lower than those in control group (P<0.05). The serum levels of inflammatory factors (TNF-α, IFN-γ, IL-33) in observation group were significantly lower than those in control group (P<0.05). The serum levels of immune indexes (IgE, EOS) in observation group were significantly lower than those in control group (P<0.05), while EOT in observation group were significantly higher than that in control group (P<0.05). The incidence of adverse reactions was 1.7%(1/59) in observation group, which was lower than 19.3%(11/57) in control group (P<0.05).Conclusion:Wenfei Zhiliudan can significantly alleviate the clinical symptoms, serum inflammatory factors and immune indicators of lung Qi deficiency cold type allergic rhinitis, with a lower incidence of adverse reactions.Keywords：allergic rhinitis;Wenfei Zhiliudan;lung Qi deficiency cold type- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect and Mechanism of Modified Buzhong Yiqi Tang and Pursed-lips Breathing on Serum TNF
-α , IL-8, IL-6, IL-1β and Cys-C in Patients at Stable Stage of COPDAbstract：Objective:To observe the effect and mechanism of modified Buzhong Yiqi Tang and pursed-lips breathing (PLB) on serum tumor necrosis factor-alpha (TNF-α), interleukin-8 (IL-8), IL-6, IL-1β and serum cystatin C (Cys-C) in patients of chronic obstructive pulmonary disease (COPD) at the stable stage.Method:Totally 120 cases of COPD treated at Affiliated Hospital of Jiangxi University of Traditional Chinese Medicine from September 2017 to March 2018 conformed to the inclusion criteria, and were randomly divided into treatment group (60 cases) and control group (60 cases). Cases of control group received Tiotropium (18 μg/ time, qd). In addition to the therapy of control group, cases of treatment group were also given Buzhong Yiqi Tang and PLB. Both groups were treated for 6 months. Percent of forced expiratory volume in one second in predicted value (FEV1%), ratio of FEV1 to forced vital capacity (FEV1/FVC), six-minute walk distance (6MWD), TNF-α, IL-8, IL-6, IL-1β and Cys-C were compared between both groups.Result:After treatment, the clinical efficacy rate of observation group was 93.33%, which was evidently higher than 86.67%of control group (P<0.05). Compared with before treatment, both groups showed decreases in TNF-a, IL-8, IL-6, IL-1β, Cys-C but increases in FEV1%, FEV1/FVC (P<0.05). Compared with control group, treatment group showed decreases in TNF-a, IL-8, IL-6, IL-1β, Cys-C but increases in FEV1%, FEV1/FVC (P<0.05).Conclusion:Buzhong Yiqi Tang and PLB has an anti-inflammatory effect on COPD by reducing levels of inflammation cytokines TNF-α, IL-8, IL-6, IL-1β and chronic inflammation markers serum Cys-C and inflammatory reaction, and alleviating airflow obstruction, promoting the levels of FEV1%, FEV1/FVC, pulmonary function control inflammatory factors, so as to improve pulmonary function and the quality of life in patients with COPD.Keywords：chronic obstructive pulmonary disease(COPD);stable period;Buzhong Yiqi Tang;pursed-lips Breathing (PLB);inflammatory cytokines;serum cystatin-C(Cys-C)- 5
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Efficacy and Mechanism of Danlu Tongdu Tablets on Lumbar Disc Herniation with Kidney Deficiency and Stasis Syndrome
Abstract：Objective:To observe short-term and long-term efficacies of Danlu Tongdu tablets on lumbar disc herniation (LDH) with kidney deficiency and stasis syndrome, and its effect on nucleus pulposus reabsorption and immunoinflammatory factors.Method:One hundred and sixty patients were randomly divided into control group (80 cases) and observation group by random number table. Patients in control group (80 cases) got acupoint massage and acupuncture, 1 time/day. In addition to the therapy of control group, patients in observation group were also given Danlu Tongdu tablets, 4 tablets/time, 3 times/days. A course of treatment was 12 weeks, and a 9-month follow-up was recorded. Lumbago and leg pain were recorded by visual simulation (VAS) before treatment and at the first, second, third month after treatment. Before and after treatment, Japanese orthopaedic association (JOA), symptoms and signs were scored. During the 9-month follow-up, relapse rate and relapse time were recorded, and VAS and JOA were scored. Absorption of nucleus pulposus was assessed, and levels of interleukin-1 (IL-1), IL-17, tumor necrosis factor-α (TNF-α), matrix metalloproteinases-3 (MMP-3) were detected.Result:By rank sum test, the clinical efficacy in observation group was better than that in control group (Z=2.125, P<0.05), and scores of JOA was better than that in control group (Z=1.924, P<0.05). At the first, second and third months after treatment, VAS scores of low back pain, leg pain were lower than those in control group (P<0.01), while the decline range were more than that in control group (P<0.01). And scores of symptoms and signs were lower than that in control group (P<0.01), whereas scores by dimensions of JOA scale and the total score of JOA were all higher than those in control group (P<0.01). Complete absorptivity of nucleus pulposus in observation group was 61.11%, which was higher than 42.03%in observation group (χ2=5.138, P<0.05). Levels of IL-1, IL-17, TNF-α and MMP-3 were lower than that in control group (P<0.01). Relapse rate in observation group was 23.61%, which was lower than 40.58%in control group (χ2=4.668, P<0.05). Average recurrence time was longer than that in control group (P<0.05), score of VAS was lower than that in control group (P<0.05), and the total score of JOA was higher than that in control group (P<0.05).Conclusion:In addition to acupuncture and massage therapy, Danlu Tongdu tablets can also be used to relieve pain and ameliorate function, improve clinical efficacy, reduce relapse rate, promote nucleus pulposus reabsorption, and relieve inflammation of nerve root.Keywords：lumbar disc herniation;kidney deficiency and stasis syndrome;Danlu Tongdu tablets;relapse rate;nucleus pulposus reabsorption;immuno-inflammatory factors- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Shenqi Compound Recipe on Intestinal Microecology of Patients with Qi and Yin Deficiency and Blood Stasis Syndrome Newly Diagnosed Type 2 Diabetes Mellitus
Abstract：Objective:To observe the effect of Shenqi compound recipe on glucose and lipid metabolism in patients with Qi and Yin deficiency and blood stasis syndrome newly diagnosed type 2 diabetes mellitus (T2DM), and its intervention effect on intestinal microecology and serum proinflammatory factors.Method:The 106 eligible patients were divided into the observation group (54 cases) and the control group (52 cases) by random number table method. Another 40 healthy volunteers in physical examination center of the hospital during the same period were enrolled as health control group. On the basis of Guidelines for the Prevention and Treatment of Type 2 Diabetes in China(2013 edition), control group was provided lifestyle interventions, such as reasonable diet, weight control, moderate exercise, salt restriction, tobacco control, alcohol restriction and psychological balance. In addition to the therapy of the control group, the observation group was given Shengi compound for oral administration, 2 times/days. Both groups were treated for 8 weeks. The fasting blood glucose (FBG), postprandial 2 h blood glucose (PBG), glycosylated hemoglobin (HbA1c), homeostasis model assessment-insulin resistance index (HOMA-IR), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) before and after treatment were evaluated. The structure and quantity of intestinal flora before and after treatment were detected. The traditional Chinese medicine(TCM)symptom was scored. The levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), IL-8, and tumor necrosis factor-α (TNF-α) were measured before and after treatment.Result:FBG, PBG, HbA1c and HOMA-IR levels in observation group were lower than those in control group (P<0.05). The operational taxonomy unit (OUT) of observation group was higher than that of control group (P<0.05). The abundances of Bacteroides and Klebsiella in observation group were higher than those in control group, while actinomyces abundance was significantly lower than that in control group (P<0.05). TC, TG and LDL-C levels in observation group were lower than those in control group, while HDL-C was higher than that in control group (P<0.05). IL-1β, IL-6, IL-8 and TNF-α levels in observation group were lower than those in control group (P<0.01). Rank-sum test showed that the efficacy on diseases of observation group was better than that of control group after treatment (Z=2.134, P<0.05).Conclusion:Shenqi compound can regulate blood glucose and blood lipid in patients with newly diagnosed T2DM (Qi and Yin deficiency and blood stasis syndrome), improve IR, intestinal microecology imbalance, and reduce non-specific inflammatory response, with a good clinical efficacy on intestinal microecology of patients with Qi and Yin deficiency and blood stasis syndrome newly diagnosed type 2 diabetes mellitus.Keywords：type 2 diabetes mellitus;Qi and Yin deficiency and blood stasis syndrome;Shenqi compound;intestinal microecology;inflammatory factor- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Clinic
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Analysis of Mechanism of Artesunate in Treatment of Liver Cancer Based on Metabolomics
Abstract：Objective:To study on the antitumor mechanism of artesunate in the treatment of liver cancer based on gas chromatography-mass spectrometry (GC-MS).Method:CellTiter-Glo® Luminescent Cell Viability Assay was used to detect activity of artesunate with different concentrations (0, 12.5, 25, 50, 100 μmol·L-1) on human liver cancer Huh7, SMMC-7721 cells for 24, 48, 72 h. GC-MS was employed to analyze the changes of metabolites of artesunate in two kinds of hepatoma cells (Huh7, SMMC-7721) for 24 h. The data was preprocessed by Postrun Analysis 4.41 workstation. Partial least squares-discriminant analysis (PLS-DA) was used to analyze two sets of differential metabolites and to analyze metabolic pathways of differential metabolites based on MetaboAnalyst 3.0 software.Result:Compared with the normal group, after two kinds of liver cancer cells was treated by artesunate, a total of 39 identical metabolites in the cells have undergone significant changes, which were mainly related to five metabolic pathways, including biosynthesis of aminoacyl-transfer RNA (tRNA), metabolism of alanine, aspartic acid and glutamic acid, metabolism of glycine, serine and threonine, metabolism of arginine and proline, metabolism of glutathione.Conclusion:Artesunate (12.5-100 μmol·L-1) can inhibit the growth of liver cancer cells (Huh7, SMMC-7721), it mainly involves five metabolic pathways, which may be the pathway of artesunate against liver cancer.Keywords：artesunate;metabolomics;liver cancer;Huh7;SMMC-7721;amino acid metabolism;energy metabolism- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Analysis of Pharmacokinetic Behavior of Five Components in Qingkailing (Lyophilized) for Injection in Normal Rats and Cerebral Ischemia Rats by UPLC-MS/MS
Abstract：Objective:To establish a UPLC-MS/MS analysis method for determination of baicalin, geniposide, chlorogenic acid, cholic acid and hyodeoxycholic acid in Qingkailing (lyophilized) for injection in rat plasma, and to investigate the pharmacokinetic behavior of this preparation in normal and cerebral ischemic rats.Method:Rats were randomly divided into normal group and cerebral ischemia model group. The rat model of cerebral ischemia was established by suture embolization. The rats were given by intraperitoneal injection, and normal saline was used as the solvent. Blood samples were taken at the corresponding time points. After treatment, UPLC-MS/MS was used to determine the blood concentration of five components. The main detection conditions were mobile phase of 0.1%formic acid aqueous solution-acetonitrile for gradient elution (0-0.25 min, 90%A; 0.25-1 min, 90%-75%A; 1-2 min, 75%-50%A; 2-2.6 min, 50%-45%A; 2.6-2.65 min, 45%-90%A; 2.65-4.0 min, 90%A), the flow rate of 0.4 mL·min-1, the column temperature at 40 ℃, electrospray ionization under negative ion mode. The pharmacokinetic parameters were fitted and the bioavailability was calculated, the differences of treatment process of five components from Qingkailing (lyophilized) for injection in normal and cerebral ischemic rats were analyzed.Result:Compared with the normal group, the area under the curve (AUC0-t) of geniposide in rats from cerebral ischemia model group decreased significantly after intraperitoneal injection of Qingkailing (lyophilized) for injection (P<0.05), and the time to peak (Tmax) of chlorogenic acid in rats from cerebral ischemia model group was significantly earlier than that in the normal group (P<0.01). Pharmacokinetic parameters of baicalin, cholic acid and hyodeoxycholic acid had no significant difference between these 2 groups.Conclusion:Qingkailing (lyophilized) for injection has a certain difference in the treatment process between normal and cerebral ischemic rats, which has certain guiding significance for the clinical treatment of cerebral ischemic diseases with this preparation.Keywords：Qingkailing (lyophilized) for injection;cerebral ischemia model;pharmacokinetics;rutin;isorhamnetin;endogenous bile acids;intraperitoneal injection- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Drug Metabolism
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Effect of Coptidis Rhizoma on Gut Microbiota of Normal and Type 2 Diabetic Rats Before and After Being Steamed with Rice Wine Based on 16S rRNA Technique
Abstract：Objective:To investigate the effect of Coptidis Rhizoma on intestinal microflora of normal and type 2 diabetic rats before and after being steamed with rice wine based on 16S rRNA sequencing technology.Method:The type 2 diabetic rat model was established by high-fat diet and streptozotocin. Normal and model rats were given the decoction of Coptidis Rhizoma or Coptidis Rhizoma steamed with rice wine every day (0.8 g·kg-1, according to raw drug quantity), the positive drug group was given metformin solution every day (0.25 g·kg-1), continuous administration for 30 d. Blood glucose was measured once a week and oral glucose tolerance was measured on the 27th day of administration. On the 30th day, feces were taken and the rats were anesthetized. Blood was taken for enzyme linked immunosorbent assay (ELISA). Colon was dissected for hematoxylin-eosin (HE) staining. Based on 16S rRNA gene sequencing technology, the alpha diversity, beta diversity, abundance and composition, and the correlation with anti-inflammatory and antioxidant activities of intestinal microflora were comprehensively analyzed and evaluated.Result:After giving normal rats Coptidis Rhizoma or Coptidis Rhizoma steamed with rice wine, there were some inflammatory reactions, imbalance of oxidation-antioxidant system, pathological damage of colon and changes of intestinal microflora, which showed some toxic and side effects, but the toxic and side effects of Coptidis Rhizoma steamed with rice wine were lower than those of Coptidis Rhizoma. Coptidis Rhizoma could reduce fasting blood glucose and improve glucose tolerance in type 2 diabetic rats before and after steaming. Although the effect of Coptidis Rhizoma steamed with rice wine had an increasing trend, there was no significant difference between before and after processing. Type 2 diabetic rats appeared mild inflammation and imbalance of oxidation-antioxidant system, characterized by significant increase of contents of interleukin (IL)-6, nuclear transcription factor (NF)-κB and malondialdehyde (MDA) (P<0.05, P<0.01), significant decrease of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) activities (P<0.01). Coptidis Rhizoma could inhibit inflammation and imbalance of oxidation-antioxidant system, and alleviate pathological damage of colon before and after steaming in type 2 diabetic tats, the effect of Coptidis Rhizoma steamed with rice wine was better than that of Coptidis Rhizoma. Significant intestinal flora disorders were observed in type 2 diabetic rats, including down-regulation of Bacteroidetes, Firmicutes and Kiritimatiellaeota, up-regulation of Proteobacteria, Actinobacteria, Acidobacteria, Chloroflexi, Cyanobacteria and Verrucomicrobia. Coptidis Rhizoma could alleviate intestinal microflora disturbance before and after steaming, and the regulation effect of Coptidis Rhizoma on Kiritimatiellaeota, Cyanobacteria and Verrucomicrobia was stronger than that of Coptidis Rhizoma steamed with rice wine. Coptidis Rhizoma steamed with rice wine could significantly reduce the level of Chloroflexi, but Coptidis Rhizoma had no such effect. The intestinal microflora may be correlated with the antioxidant capacity of Coptidis Rhizoma before and after steaming.Conclusion:Coptidis Rhizoma can treat type 2 diabetic rats by regulating intestinal microflora before and after steaming with rice wine, and the effect of Coptidis Rhizoma steamed with rice wine is better. The toxic and side effects of Coptidis Rhizoma on intestinal flora are reduced by steamed with rice wine. The processing mechanism of increasing efficacy and decreasing toxicity of Coptidis Rhizoma steamed with rice wine is closely related to intestinal microflora.Keywords：Coptidis Rhizoma;raw products;steamed products;16S rRNA technique;type 2 diabetes mellitus;intestinal microflora;antioxidant activity- 6
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Establishment of Determination of Fractal Dimension of Ethanol-precipitated Flocs of Two Root Medicinal Herbs
Abstract：Objective:To establish a method for the determination of fractal dimension of ethanol precipitation flocs in two kinds of root medicinal herbs (Astragali Radix and Isatidis Radix).Method:The ethanol-precipitated flocs of Astragali Radix and Isatidis Radix were taken as the research objects. Firstly, the original microscopic images of the ethanol precipitation flocs were binarized by Ostu algorithm. Secondly, the influence of sampling volume, dilution multiple, image magnification ratio and number of image acquisition sites on fractal dimension was investigated using perimeter-area method.Result:The ethanol precipitation flocs of Astragali Radix and Isatidis Radix had fractal characteristics. At the same time, the stable and reliable fractal dimension of the flocs could be obtained when the sampling volume was 600-800 μL, the sample was diluted to 5-20 times, the microscopic image was enlarged to 200 times or 400 times, and four different image acquisition sites were selected.Conclusion:The established method is stable and feasible, which can provide reference for the fractal theory of ethanol precipitation flocs of root medicinal herbs.Keywords：fractal dimension;ethanol precipitation;flocs;Astragali Radix;Isatidis Radix;microscopic images;granules- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Gastrointestinal Regulation of Red Yeast Rice on Mice with Syndrome of Food Retention Due to Spleen Deficiency Before and After Fermentation
Abstract：Objective:To investigate gastrointestinal regulation of red yeast rice on spleen deficient dyspepsia mice, and provide reference for clarifying mechanism of invigorating spleen to promote digestion of fermented traditional Chinese medicine.Method:Mice were randomly divided into 5 groups, including the blank group, the model group, the unfermented red yeast rice treatment group (1.17 g·kg-1·d-1), the fermented red yeast rice treatment group (1.17 g·kg-1·d-1), the domperidone group (3.9 mg·kg-1·d-1), ten mice in each group were continuous intragastric administration for 10 days. The effects of red yeast rice before and after fermentation on the general behavior, body weight, food intake and gastrointestinal function of spleen deficient dyspepsia mice were compared. The effects of red yeast rice on the gastrointestinal hormone [motilin (MTL), gastrin (GAS), 5-hydroxytryptamine (5-HT), vasoactive intestinal peptide (VIP)] levels in serum of spleen deficient dyspepsia mice were determined by enzyme-linked immunosorption assay (ELISA). The regulation function of red yeast rice on the pathological changes of gastrointestinal tissue of spleen deficient dyspepsia mice was investigated by hematoxylin-eosin (HE) staining. The effect of red yeast rice on gut microbiota of spleen deficient dyspepsia mice was investigated, the primers were designed on bacterial 16S rRNA V3-V4 region sequences and Illumina Miseq platform was used for high-throughput sequencing.Result:Compared with the model group, fermented red yeast rice could recover the body weight and food intake, reduce gastric residual rate (P<0.01) and enhance the small intestinal propulsive rate (P<0.01) of spleen deficient dyspepsia mice. Red yeast rice could increase the gastrointestinal hormone levels of MTL and 5-HT, while reduce the VIP level of spleen deficient dyspepsia mice. Red yeast rice could restore the light congestion and local structural disorder glands of gastric mucosa of spleen deficient dyspepsia mice. Red yeast rice could restore the pathological changes of intestinal tissue of spleen deficient dyspepsia mice by means of increasing the length of intestinal villus and the thickness of mucosa. Red yeast rice could restore the structure of the gut microbiota of spleen deficient dyspepsia mice by means of reducing the relative abundance of pathogenic microorganisms such as Escherichia-Shigella, while increasing the relative abundance of the beneficial bacteria such as Lactobacillus and Bifidobacterium.Conclusion:Fermented red yeast rice can enhance the gastrointestinal function of spleen deficient dyspepsia mice, the mechanism may be related to regulating gastrointestinal hormone level, improving histopathology of gastrointestinal tissue and restoring intestinal flora structure.Keywords：red yeast rice;fermentation;syndrome of food retentiondue to spleen deficiency;gastrointestinal regulation;gastrointestinal hormones;gastrointestinal tissue morphology;gut microbiota- 11
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Grade Evaluation of Magnoliae Officinalis Cortex Processed with Ginger Juice Based on Relative Quality Constant
Abstract：Objective:To establish the grade evaluation standard for Magnoliae Officinalis Cortex processed with ginger juice by combining traditional morphology evaluation with modern intrinsic quality evaluation.Method:The morphological parameters and contents of intrinsic pharmacodynamic index components of 28 batches of Magnoliae Officinalis Cortex processed with ginger juice were determined, and the relative quality constants were calculated. Assuming that the average relative quality constant was 100%, more than 120%of the samples were classified as the first grade, 60%to 120%as the second grade, the remaining as the third grade.Result:The relative quality constant of Magnoliae Officinalis Cortex processed with ginger juice ranged from 0.09 to 1.78. The relative quality constant of the first grade was ≥0.64, the second grade was 0.32-0.64, while the third grade was ≤0.32.Conclusion:Relative quality constant combines external indexes of traditional morphology and internal indexes of pharmacodynamic components, which can objectively, reasonably and scientifically classify the grade of Magnoliae Officinalis Cortex processed with ginger juice, and provide reference for establishing and improving the grade evaluation standard of this decoction pieces.Keywords：Magnoliae Officinalis Cortex processed with ginger juice;decoction pieces;relative quality constant;grades evaluation;magnolol;honokiol;appearance- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Pharmacy and Processing
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New Phenolic Glycoside from Aerial Part of Allium sativum
Abstract：Objective:To study the chemical constituents from the aerial part of Allium sativum.Method:The aerial part of A. sativum was treated by heating reflux extraction with 70%ethanol. The combined extracting solution was treated with decompressed recovery solvent to obtain the total extract. The total extract's thick paste was suspended with an appropriate amount of distilled water, and then extracted with petroleum ether, ethyl acetate and water-saturated n-butyl alcohol to obtain petroleum ether extraction fraction, ethyl acetate extraction fraction and n-butyl alcohol extraction fraction. The n-butyl alcohol extraction fraction was isolated and purified by positive phase silica gel column chromatography, ODS column chromatography and preparative HPLC, etc. The structures of the chemical constituents were elucidated by means of physicochemical properties and spectroscopic analysis, and spectral data, like 1H-NMR, 13C-NMR, DEPT, HMBC and HR-ESI-MS.Result:Seven compounds were isolated and identified as 2-methoxy-phenol-1-O-α-L-arabinopyranosyl-(1→6)-β-D-glucopyranoside(1), phenyl-ethanol-4-O-β-D-xylopyranosyl-(1→6)-β-D-glucopyranoside(2), trans-resveratrol 3-O-β-D-glucopyranoside(3), stigmasterol(4), β-sitosterol(5), stigmasterol-3-O-β-D-glucopyranoside(6) and daucosterol(7).Conclusion:Compound 1is a new phenolic glycoside named dasuanxinoside J, and compounds 2-7 are isolated from A. sativum for the first time.Keywords：Allium sativum;aerial part;phenolic glycosides;dasuanxinoside J;phytosterols- 6
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Chemical Constituents in Endophytic Fungus
Nigrospora oryzae from CordycepsAbstract：Objective:To study the chemical constituents in theendophytic fungus Nigrospora oryzae from Cordyceps.Method:It was cultured with brownrice, and isolated and purified by chromatographic procedures, and the compounds were identified by NMR, ESI-MS and other spectral methods.Result:Totally 15 compounds were identified as mellein (1), linoleic acid (2), 2-(2-hydroxyethyl)phenol (3), (3R)-mellein methyl ether (4), (3R, 4S)-4-hydroxymellein (5), 2-hydroxybenzaldehyde (6), 3-phenylpropane-1, 2-diol (7), 1-phenyl-1, 2-ethanediol (8), cyclo-(R-Prommmm-S-Ile) (9), cyclo-(D-Pro-L-Leu) (10), 2-hydroxybenzaldehyde (11), 4-hydroxy-8-O-methylmellein (12), cyclo-(S-Pro-S-Phe) (13), cyclo-(D-Pro-L-Ile) (14), cyclo-(D-Pro-L-Leu) (15).Conclusion:Compounds 1-15were isolated from this fungus for the first time.Keywords：Cordyceps;Nigrospora oryzae;endophytic fungus;mellein- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Chemical Constituents of Ethyl Acetate Extract from Seed Cake of Camellia oleifera and Its Anti-inflammatory Activities
Abstract：Objective:To intensively study the chemical constituents from the seed cake of Camellia oleifera and its pharmacological activities, in order to provide scientific basic for its further development and utilization.Method:All kinds of column chromatography and spectral methods were employed to isolate and identify the monomeric compounds from its ethyl acetate portion of ethanol extract. The in vitro anti-inflammatory effects were evaluated by LPS-induced inflammatory model in RAW264.7 macrophages.Result:Eight phenolic acids and two flavonoids were isolated from the ethyl acetate soluble portion and identified as p-hydroxybenzoic acid(1), protocatechuic acid(2), gallic acid(3), methyl gallate(4), ethyl gallate(5), isovanillic acid(6), ethyl 3, 4-dihydroxylbenzoate(7), 2-(3′, 4′-dihydroxyphenyl)-1, 3-benzodioxole-5-aldehyde(8), quercetin(9), rutoside(10). Among them, compounds 4-8were first isolated from this plant. These compounds had good anti-inflammatory activities against NO production in LPS-stimulated RAW264.7 macrophages in an obvious dose-dependent manner. Among them, compound 8showed a strongest activity.Conclusion:The above results show that the phenolic acids and flavonoids from seed cake of C. oleifera have good prospects for the development and application of anti-inflammatory drugs.Keywords：Camellia oleifera;seed cake of Camellia oleifera;ethyl acetate extract;2-(3′, 4′-dihydroxyphenyl)-1;3-benzodioxole-5-aldehyde- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Chemical Constitutes of Cardiac Glycosides in
Scurrula parasitica and ItsNerium indicum by UPLC-Q-TOF-MS/MSAbstract：Objective:To establish a method for identifying cardiac glycosides in Scurrula parasitica and its Nerium indicum host by UPLC-Q-TOF-MS/MS. With safflower parasitoids with sweet-scented osmanthus trees as the host and their host osmanthus tree samples used for control, the chemical constituents of the cardiac glycosides were identified by comparison between the cardiac glycoside reference substances and literatures, so as to analyze the correlation between the safflower parasitoid and its host oleander glycoside components, and evaluate the host's impact on the quality of Taxilli Herba.Method:Samples of S. parasitica (parasitic on N. indicum and Osmanthus fragrans), N. indicum and O. fragrans were collected. Samples of S. parasitica and its O. fragrans host were taken for control. All of the samples were extracted through ultrasonic extraction with 70%ethanol. ACQUITY UPLC HSS T3 C18(2.1 mm×100 mm, 1.8 μm) column was adopted with mobile phase A comprising 0.1%formic acid water and mobile phase B comprising acetonitrile for gradient elution. The sample size was 0.5 μL. The flow rate was 0.6 mL·min-1. The column temperature was maintained at 40 ℃. MassLynx V4.1 software was used to analyze the data. Identification and correlation of chemical constitute of cardiac glycosides in S. parasitica and its N. indicum host were performed through analysis on cardiac glycosides reference substances, relevant literatures, elemental composition of compounds and positive and negative ion mode mass spectrometry data.Result:A total of 26 compounds of cardiac glycoside were identified, including 25 compounds of cardiac glycoside from N. indicum host, and 5 compounds of cardiac glycoside from S. parasitic(parasitic on N. indicum). none of cardiac glycosides were found in S. parasitica (parasitic on O. fragrans ) and its O. fragrans host.Conclusion:It was rapid, accurate and comprehensive to identify cardiac glycosides in S. parasitica (parasitic on N. indicum) and its N. indicum host by UPLC-Q-TOF-MS/MS. S. parasitica itself does not contain cardiac glycosides, its host may impact the quality of S. parasitica by delivering cardiac glycosides, a kind of its characteristic compound.Keywords：Scurrula parasitica;Nerium ndicum;cardiac glycoside;host;Osmanthus fragrans- 5
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Cloning and Prokaryotic Expression of Squalene Epoxidase Gene from
Panax vietnamensis var.fuscidiscus Abstract：Objective:To clone the squalene epoxidase genes of Panax vietnamensis var. fuscidiscus(PvfSE), and perform bioinformatics analysis and prokaryotic expression.Method:Total RNA was extracted from root of P. vietnamensis var. fuscidiscus by trizol method, and reverse-transcribed into first stand of cDNA. Specific primers for PvfSE cloning were designed according to the transcriptome data of P. vietnamensis var. fuscidiscus, and the cDNA sequence of PvfSE gene was isolated. Bioinformatics of PvfSE was analyzed by relevant software. The prokaryotic expression vector pMal-c2X-PvfSE was built to express recombinant protein in Escherichia coli cells.Result:The PvfSE gene contained a 1 887 bp open reading frame, encoding a predicted protein of 628 amino acids. The calculated molecular weight was 68.8 kDa, the theoretical isoelectric point was 9.28, the aliphatic index was 95.18, the grand average of hydropathicity was -0.060, and the instability index was 40.36. The protein was unstable. Bioinformatics analysis showed that PvfSE had two transmembrane domains and no signal peptide. PvfSE was most likely to be located in chloroplast or cytoplasmic membrane. PvfSE was a mixed protein with FAD/NAD(P) binding domain and squalene epoxidase domain. Sequence alignment and phylogenetic analysis demonstrated that PvfSE had a relatively close relationship with CpSE1, CpSE3, OsSE1 and OsSE2, which was involved in the biosynthesis of triterpene saponins in Cucurbita pepo and Ononis spinosa. In addition, PvfSE protein was expressed in E. coli.Conclusion:In this study, PvfSE gene was cloned and expressed in BL21(DE3), which lays a foundation for the further study on the gene functions of PvfSE and the biosynthetic pathway of triterpenoid saponins in P. vietnamensis var. fuscidiscus.Keywords：Panax vietnamensis var. fuscidiscus;squalene epoxidase;gene cloning;bioinformatics analysis;prokaryotic expression- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Grade Standard of Glycyrrhizae Radix et Rhizoma Pieces
Abstract：Objective:On the basis of traditional experience identification, and appearance characteristics and intrinsic index components of Glycyrrhizae Radix et Rhizoma pieces, a comprehensive evaluation was carried out to explore the method of grading Glycyrrhizae Radix et Rhizoma pieces and establish grading standards.Method:Based on the investigation of 44 batches of Glycyrrhizae Radix et Rhizoma pieces, including their properties, glycyrrhizin content and ammonium glycyrrhizinate content, Glycyrrhizae Radix et Rhizoma pieces were classified according to the market grading to establish the grading standards.Result:Based on the above data, the Glycyrrhizae Radix et Rhizoma pieces were classified into four grades. First-class: average diameter>1.66 mm, average weight>0.54 g, glycyrrhizin content>1.10%, glycyrrhizic acid content>2.12%. Second-class: the average diameter is between 1.40-1.66 mm, the average weight is between 0.42-0.54 g, the content of glycyrrhizin is between 0.74%-1.10%, and the content of glycyrrhizic acid is between 1.95%-2.12%. Third-class: the average diameter is between 1.07-1.40 mm, the average weight is between 0.29-0.42 g, the content of glycyrrhizin is between 0.65%-0.74%, and the content of glycyrrhizic acid is between 1.88%-1.95%. Substandard: the average diameter<1.07 mm, average weight<0.29 g, content of glycyrrhizin<0.65%, content of glycyrrhizic acid<1.88%.Conclusion:This experiment combines the traditional experience and modern analysis method, in order to develop a scientific, reasonable and accurate classification method.Keywords：Glycyrrhizae Radix et Rhizoma pieces;diameter;weight;glycyrrhizin;glycyrrhizic- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Classification and Identification of Cynomorii Herba from Different Producing Areas Based on Fourier Rransform Infrared Spectroscopy and Chemometrics
Abstract：Objective:To realize the classification and identification of Cynomorii Herba from different producing areas based on fourier transform infrared spectroscopy (FTIR) and chemometrics.Method:FTIR spectrum data of 106 batches of Cynomorii Herba from 12 cities in 5 provinces were collected by transmission method and preprocessed. The FTIR fingerprints of Cynomorii Herba were established, and spectrum analysis was performed. The FTIR similarities of Cynomorii Herba from different producing areas were calculated by correlation coefficient method. The first derivative (1D) spectrum of average FTIR of Cynomorii Herba from different producing areas were obtained. The soft independent modeling of class analog (SIMCA) model based on principal component analysis (PCA) was established by the preprocessed 1D spectrum data. The orthogonal partial least squares (OPLS) model was established by top 6 principal components.Result:The FTIR fingerprint trend and main absorption peaks of Cynomorii Herba from different producing areas were basically the same, and 16 common characteristic absorption peaks were recognized. Similarity and 1D spectrum of FTIR fingerprint of Cynomorii Herba from different producing areas showed significant and unique characteristics. The established SIMCA model can realize the classification and identification of Cynomorii Herba from different provinces, while OPLS model can realize accurate classification and identification of Cynomorii Herba in different cities. The classification and identification of Cynomorii Herba from 12 city producing areas showed obvious geographical clustering characteristics.Conclusion:The established method based on FTIR and chemometrics can realize the classification and identification of Cynomorii Herba from 12 cities.Keywords：Cynomorii Herba;fourier transform infrared spectroscopy;first derivative spectrum;soft independent modeling of class analog;orthogonal partial least squares;origin identification- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Resource and Quality Evaluation
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Mechanism of Modified Taohe Chengqitang in Treating Type 2 Diabetes Mellitus Based on Network Pharmacology
Abstract：Objective:To study the potential therapeutic targets of modified Taohe Chengqitang for type 2 diabetes mellitus based on network pharmacology.Method:Based on TCMSP database and Uniprot database, the active constituents and target genes of flavored modified Taohe Chengqitang were screened.Target genes of type 2 diabetes were screened by gene cards database and OMIM database, and Cytoscape software was used to construct " active component-target" interaction network diagram.The active component targets and disease targets were uploaded to the STRING database, the protein interaction network map (PPI) was constructed, and the characteristic values were calculated, and core genes were screened by using R language.Finally, R language was used to analyze gene ontology (GO) enrichment and kyoto encyclopedia of genes and genome (KEGG) pathway enrichment of key targets.Result:The 155 active components and 106 effective targets of modified Taohe Chengqitangin the treatment of type 2 diabetes were predicted.Quercetin, kaempferol and isorhamnetin were the most effective components, while estrogen receptor alpha (ESR1), peroxidosomal hyperplasia activates receptor gamma (PPARG) and androgen receptor (AR) were the most effective components.Core genes in PPI network areepidermal growth factor receptor (EGFR)and ESR1, etc.GO enrichment analysis shows that it can affect gene transcription, nuclear receptor activity, hormone receptor binding, neurotransmitter, etc.Enrichment analysis of KEGG pathway showed that fluid shear stress and atherosclerosis pathways were the most significant pathways, followed by advanced glycation end products-receptor for AGE (AGE-RAGE) pathway.Conclusion:Predicted by the method of network pharmacology modified Taohe Chengqitang key targets for prevention and treatment of type 2 diabetes and related pathways, results suggest the recipe has multiple targets, multiple pathways, such as complex mechanism, not only show that modified Taohe Chengqitang has hypoglycemic effect, but it also has anti-inflammatory, improve insulin resistance, regulate lipid metabolism, and biological functions.Keywords：modified Taohe Chengqitang;type 2 diabetes;network pharmacology;potential targets- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Construction of Prescription Database of
The Four Medical Tantras and Analysis of Its Medication Rules Based on Complex Network Analysis and Visual FoxProAbstract：Objective:The Four Medical Tantras is a classic work of Tibetan medicine, in which drug treatment is one of the four main treatment methods, and prescriptions are the main form of clinical drug use. The study on the medication rules of The Four Medical Tantras has positive significance for the clinical medication research of Tibetan medicine.Method:Prescriptions recorded in Secret Medical Code and Follow-up Medical Code from The Four Medical Tantras were collected, the prescription database was constructed by Visual FoxPro 9.0, combined the dual perspectives of the " fifteen categories" (skabs bco lnga) of Tibetan medical diseases and the " eighteen practices" (lag len bco brgyad) of treatment, and Gephi 0.9.2 complex network was used to analyze its medication rules.Result:The Four Medical Tantras contained 2 695 Tibetan medicine prescriptions, average number of drugs in each prescription was 6.6, prescription composed of ≤7 drugs accounted for 69.2%of the total prescription. There were 1 002 kinds of medicines involved in the total prescription, of which 156 was the nodal degrees of Chebulae Fructus, Bambusae Concretio Silicea, Carthami Flos, Piperis Longi Fructus and Zingiberis Rhizoma and so on; followed by 91 nodal degrees of butter (mar), sugar, barley sweetener (sbang ma), honey and brown sugar and so on, which were Tibetan medicinal and edible medicines. According to the classification of diseases in the fifteen categories, there were 495 prescriptions for traumatology (mtshon rma), 323 for fever (tsha ba), 278 for toxics and 197 for three factors diseases. According to the treatment method of eighteen practices, 162 powder, 77 decoction, 84 laxative (bxhal) and others were used. Association rule analysis found that edge weights of Borneolum Syntheticum, Bambusae Concretio Silicea, Carthami Flos, Santali Albi Lignum and other cold medicines were 59, edge weights of pomegranate seeds, Piperis Longi Fructus, Cinnamomi Cortex, Amomi Fructus Rotundus and other warm medicines were 45.Conclusion:The Four Medical Tantras has some characteristics, including the medication rules of elliptic philosophy with cool-warm medicinal properties as the dual core, elimination-diarrhea (zhi-sbyong) as medication methods, weight of rlung-mkhris pa-bad kan as medication dialectical thinking and so on. Among them, the clinical application of butter, barley sweetener, sugar and other special delivery agents of Tibetan medicine is prominent. The prominent clinical application of special Tibetan medicines embodies the philosophy of Tibetan culture with the emphasis on overall balance. It has the multi-dimensional medication rule of " diet plus medicine, medicine plus therapy" , and has positive significance for revealing the research on medication rules of Tibetan medicine.Keywords：The Four Medical Tantras;Tibetan medicine;prescription;database;medication rule;complex network;fifteen categories;eighteen practices- 11
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Data Mining
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Natural Law and Pathogenesis Based on Wufang Circular Motion
Abstract：Various academic discussions have been officially proposed in the " theory of circular motion" in Circular Motion of Ancient Chinese Medicine. The theory of circular motion reflects the inheritance and development of the theory of Yin-Yang and five elements in ancient Chinese medicine, which has a great significance in the modern basic theory research and clinical practice of traditional Chinese medicine. On the basis of ancient literatures, this article aims to discuss the natural law of circular motion from the perspectives of the path of sun and 24-solar-terms of seasons via five-elements and Wufang circular motion theory according to circular motion. The circular motion has an effect on human body and natural environment. The invasion of pathogen is caused by deficiency of essential Qi. The combination of the healthy-Qi deficiency with the deficiency-type pathogen leads to the occurrence of external contraction. Therefore, the abnormality of Yang-Qi including nature and human is the main cause of the disease's development. Because the circular motion can expound the change rule and mutual relations of nature and human, we can predict the development of disease by it. When the circulation motion is disordered, clinical symptoms appear with inner pathogenesis. The understanding of the change of the circulation motion is helpful for us to have a deeper realization of the disease, grasp the pathogenesis and provide treatment based on syndrome differentiation. The authors explain nature environment and human physiology, and elaborate the relations between natural rule and Wufang's effect on circulation motion, in order to provide certain suggestions to guide clinical treatment based on pathogenesis.Keywords：Wufang circular motion;Wufang pathogenesis;Zhou Yi;Yifa Fangyi Lun;Qi activities- 13
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Textual Research on Processing Methods of Pinelliae Rhizoma in Successive Dynasties
Abstract：This paper discussed the processing methods of Pinelliae Rhizoma in the successive dynasties by consulting ancient and modern books, literature and related codes. The processing methods of Pinelliae Rhizoma has experienced various historical periods. Since the processing method of Pinelliae Rhizoma processed by slicing appeared in the Huangdi Neijing, there have been a series of different processing methods and requirements are founded, such as cleansing, cutting, decocting and so on. Among them, the raw Pinelliae Rhizoma, Pinelliae Rhizoma Praeparatum, Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine, Pinelliae Rhizoma Praeparatum cum Alumine are still in use today and are widely used. However, there are differences in the processing methods, accessories and their dosages of these various processed products in different dynasty. The processing technology and quality standard of Pinelliae Rhizoma need to be further standardization.Keywords：processing methods;Pinelliae Rhizoma;Pinelliae Rhizoma Praeparatum;Pinelliae Rhizoma Praeparatum cum Zingibere et Alumine;Pinelliae Rhizoma Praeparatum cum Alumine;textual research;historical evolution- 11
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Academic Discussion
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Effect of Traditional Chinese Medicine in Regulating Immune Mechanism of Chronic Urticaria
Abstract：Chronic urticaria (CU) is one of common diseases of dermatology. It has the clinical characteristics of allergy, multiple, recurrent and persistent recovery. The clinical symptoms of patients are mainly pruritus and wheal, and concurrent with vascular edema, some patients may even suffer from chest tightness, nausea and other systemic symptoms. Its pathogenesis is still unclear, and it is generally considered to be related to autoimmune, chronic infection, coagulation mechanism, vitamin D deficiency and so on. Among them, autoimmunity may be one of the important causes for chronic urticaria. Clinical studies mostly focus on immunoglobulin E, T cell subsets CD4+ T cells, CD8+ T cells, cytokines interleukin-2, interferon-γ, and complement C3, C4. Western medicine therapies mostly use antihistamines. The first generation of antihistamines have an obvious inhibitory effect on central nervous system, and side reactions, such as sedative and anticholinergic Effect. Although the second generation has a reduced inhibitory effect on central nervous system, it is easy to cause heart toxicity and relapse after drug withdrawal. Therefore, it is particularly important to find a safe and effective traditional Chinese medicine(TCM) therapy. Urticaria belongs to the category of " hidden rashes" in TCM. The etiology and pathogenesis are insufficient endowment of body elements, insufficient external defense or improper diet, damp-heat connotation. Classic prescriptions and self-prepared formulas, such as Jade Screen Powder, shall be used in the treatment of phlegm and blood stasis, with a clear clinical effect and a low recurrence rate. In addition, by reducing the level of immunoglobulin E in patients, improving CD4+ T cells, CD8+ T cell levels, regulating the levels of related cytokines, increasing the levels of complement C3, C4, these therapies could reduce IgE levels, release of anaphylatoxins, stimulation of mast cells and inflammatory reactions, and regulate the body's immune function, with a satisfactory efficacy. Therefore, the author has summarized the clinical researches on the mechanism of traditional Chinese medicine in regulating chronic urticaria in past five years.Keywords：chronic urticaria;immune mechanism;traditional Chinese medicine (TCM);regulate- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Modern Pharmacological Effect and Mechanism of Shengmai San
Abstract：Shengmai San was first recorded in Medical Origin and Insights, a medical book written by ZHANG Yuan-su. Shengmai San composed of Ginseng Radix et Rhizoma, Ophiopogonis Radix and Schiandrae Chinensisin Fructus. Ginseng Radix et Rhizoma as the monarch drug in the prescription has effects in invigorating vigour potently, and promoting production of body fluid to quench thirst. Ophiopogonis Radix as a sweet-cold minister drug has effects in nourishing yin, clearing heat and generating fluid and moistening lung to stop cough in the prescription. Both medicines are combined to have a good effect in replenishing Qi. Schiandrae Chinensisin Fructus is an adjuvant medicine, with effects in acid astringency, retaining Yin with astringent and hidroschesis. Ginseng Radix et Rhizoma has effects in strengthening the healthy energy, and Ophiopogonis Radix has a effect in retaining yin with astringent. It is a classic prescription for treating deficiency of Qi and Yin. With the in-depth research of modern medical experts, it is found that Shengmai San not only has a significant therapeutic effect on cardiovascular diseases, central nervous system diseases and endocrine system diseases, but also a good effect on digestive system diseases, immune system and hematopoietic system diseases and shock. As China's aging population, irregular diet and other problems become increasingly serious, the incidences of cardiovascular diseases, senile dementia, cerebral infarction, diabetes, liver dysfunction and other diseases have become higher and younger, which is a serious threat to human health. Therefore, by consulting a large number of domestic literatures, the authors respectively elaborated the therapeutic effect and mechanism of Shengmai San and its modified prescriptions on the above diseases, with the aim to reveal the target and mechanism of this prescription on the above diseases, provide theoretical basis for better treatment of the above diseases and promote the clinical application of this prescription. In addition, it provides reference ideas for the research of other classical famous formula.Keywords：Shengmai San;myocardial ischemia;cerebral ischemia;diabetes mellitus;liver protection;anti-shock- 13
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Research Progress on Effect of Hypoglycemic Mechanism of Berberine in Treatment of Type 2 Diabetes Mellitus
Abstract：In recent years, type 2 diabetes mellitus (T2DM) has an increasing incidence worldwide along with the improvement of people's living standards, and emerged as a serious threat to human health. T2DM is a progressive metabolic disease characterized by hyperglycemia, and its routine therapies include diet control, the use of oral hypoglycemic drugs or subcutaneous injection with insulin. At present, in addition to chemical drugs, such as metformin and thiazolidinediones, researchers have also found that natural medicines and traditional Chinese medicine compounds have mild hypoglycemic and insulin sensitizing effects. Besides, these drugs also have effects in alleviating diabetes complications and maintaining glucose homeostasis. Berberine is an isoquinoline alkaloid mainly isolated from Coptis chinensis with multiple pharmacological activities. Currently, berberine is considered to be one of the most promising natural hypoglycemic agents for the treatment of type 2 diabetes mellitus. However, mechanisms of the hypoglycemic effect of berberine were complex. In this review, we summarized the pharmacological mechanisms of hypoglycemic effect of berberine, including improving insulin resistance, promoting insulin and glucagon-like peptide-1 (GLP-1) secretion, inhibiting hepatic gluconeogenesis, reducing glucose absorption in intestinal cells, suppressing oxidative stress and inflammation and modulating gut microbiota, clarified the mechanisms of hypoglycemic effect of berberine could help to understand the pharmacology of berberine in the treatment of diabetes mellitus and provided evidence for rational application of berberine in the treatment of type 2 diabetes.Keywords：berberine;type 2 diabetes;hypoglycemic activity;research progress- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Advances in Research on Mechanism of Salvianolic Acid B Against Oxidative Stress
Abstract：Salvia miltiorrhiza Bge was first published in Shennong Bencaojing. Salviae Miltiorrhizae Radix et Rhizoma has the effect of promoting blood circulation, cooling blood and eliminating phlegm. Modern pharmacological studies have found that Salviae Miltiorrhizae Radix et Rhizoma has strong heart, improve microcirculation and antithrombotic effects. The decoction of Salviae Miltiorrhizae Radix et Rhizoma is commonly used in clinical practice. The dissolved component of Salviae Miltiorrhizae Radix et Rhizoma is mainly salvianolic acid. Among them, salvianolic acid B is one of the water-soluble components of Salviae Miltiorrhizae Radix et Rhizoma. It has strong physiology. Activity, salvianolic acid B is the strongest antioxidant activity of salvianolic acids. Salvianolic acid B regulates nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, decoupled endothelial nitric oxide synthase (eNOS) in vivo, and regulates Janus protein tyrosine kinase 2/signal transducer and transcriptional activation 3/vascular endothelial growth factor (JAK2/STAT3/VEGF) and tumor suppressor(p53), B cell lymphoma-2(Bcl-2), Bcl-2-related X protein (Bax), Caspase-3 anti-apoptotic pathway, inhibition of extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK), c-Jun N-terminal kinase/mitogen-activated protein kinase (JNK/MAPK), mitogen-activated protein kinase p38 (p38MAPK) and estrogen receptor alpha/phosphatidylinositol 3-kinase/serine-threonine protein kinase/nuclear transcription factor kappa B (ERα/PI3K/Akt/NF-κB) anti-inflammatory pathway, silencing information regulator 2 related enzyme 1/nuclear factor E2 related factor 2/Kelch-like ECH combined protein 1-antioxidant response element (SIRT1/Nrf2/Keap1-ARE) and NAD-dependent deacetylase sirtuin Forkhead transcription factor 1/superoxide dismutase on-3/β cells (NAD-dependent deace tylase sirtuin-3, SIRT3/FOXO1/SOD2) antioxidant pathway and autophagy pathway inhibit oxidative stress and protect tissues and organs from oxidative damage. Based on the study of the pathway of salvianolic acid B against oxidative stress injury, this paper reviews the related research published in domestic and foreign journals in the past five years, and provides new ideas for expanding the clinical application range of salvianolic acid B.Keywords：salvianolic acid B;oxidative stress;signaling pathway;research progress- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Review
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