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Issue 10 ， 2020
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Effect of Danggui Shaoyaosan Ameliorates in Alleviating H2O2-induced SH-SY5Y Cells Oxidative Stress Through Modulation of NF-
κ B PathwayAbstract：Objective:To investigate the regulatory effect of Danggui Shaoyaosan (DSS)-containing serum on oxidative stress and inflammation in H2O2-induced SH-SY5Y cells.Method:Methyl thiazolyl tetrazolium(MTT) assay was used to determine the cell activity and construct the H2O2-induced cell damage model with the optimal time and dose. Normal group, model group and high, medium and low-dose DSS groups(2.5%, 5%, 10%) were set up. MTT method was used to detect cell activity, spectrophotometry anti-oxidation indexes of malonaldehyde (MDA), catalase (CAT) and superoxide dismutase (SOD) and glutathione (GSH). Real-time quantitative PCR (Real-time PCR) was used to detect tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) mRNA expressions. And immunofluorescence test was adopted to detect nuclear transcription factor-κB(NF-κB) p65 nuclear translocation of the DSS after the intervention.Result:After 24 h intervention with 250 μmol·L-1 H2O2, SH-SY5Y cell viability was about 55%, which was the best modeling condition. After high, medium and low-dose DSS intervention on H2O2-damaged cell model, compared with the model group, the cell activity showed a dose-dependent increase (P<0.05), MDA was significantly reduced (P<0.05), and antioxidant indexes CAT, SOD and GSH were significantly increased (P<0.05). H2O2 could significantly increase SH-SY5Y cell inflammatory factor TNF-α, IL-6 and IL-1β mRNA expressions, and promote activation of cytoplasmic NF-κB and nuclear translocation. DSS-containing serum showed a dose-dependent inhibition of NF-κB p65 from nuclear, and reduced inflammatory factor levels, such as TNF-α, IL-6 and IL-1β.Conclusion:DSS-containing serum can significantly reduce the oxidative damage in H2O2-induced SH-SY5Y cells by improving their antioxidant status, and reduce the inflammatory response by inhibiting NF-κB signaling pathway.Keywords：Danggui Shaoyaosan;oxidative stress;nuclear transcription factor-κB(NF-κB);inflammation- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Wumeiwan on Intestinal Microflora, Inflammatory Factor and Short Chain Fatty Acids in Type 2 Diabetic Rats
Abstract：Objective:To study the effects of Wumeiwan on blood glucose, intestinal microflora, tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10) and dietary fiber fermented by intestinal microflora in type 2 diabetic (T2DM) rats.Method:Totally 80 SD clean rats were selected as experimental subjects, and 10 of them were randomly selected as the normal group. The remaining 70 rats were given high-sugar and high-fat emulsion for 8 weeks and intraperitoneally injected with streptozotocin (STZ, 35 mg·kg-1) to establish the rat model of T2DM. The fasting blood glucose higher than 11.10 mmol·L-1 was considered as a successful model. The rats that were not successfully modeled were removed, and the remaining rats that were successfully modeled were randomly divided into model group, metformin group, high-dose Wumeiwan group, medium-dose Wumeiwan group and low-dose Wumeiwan group, with 10 rats in each group. Normal group and model group received (ig) normal saline (20 mL·kg-1·d-1), while metformin group (ig metformin 200 mg·kg-1·d-1), Wumeiwan high, medium and low dose groups (ig Wumeiwan 20, 10, and 5 g·kg-1·d-1) received corresponding drugs respectively. Blood glucose and body weight of rats were monitored regularly before and after administration of the drugs. Blood and feces were collected after four weeks of administration.16S-rDNA high-throughput sequencing technology was used for gene sequencing of intestinal flora. Enzyme linked immunoassay (ELISA) was used to detect the serum levels of inflammatory factors TNF-α and IL-10 in rats, and the contents of acetic acid, propionic acid and butyric acid in feces were detected by gas chromatography.Result:As compared with the normal group, the body weight decreased significantly (P<0.01). Bacteroidetes, Actinobacteria, Bacteroides, Clostridium increased, Firmicutes, Deltaproteobacteria, and Lactobacillus decreased, fasting blood glucose and serum TNF-α levels increased significantly (P<0.01), IL-10 level decreased (P<0.01), and the contents of acetic acid, propionic acid and n-butyric acid of short-chain fatty acids decreased(P<0.05, P<0.01) in model group. As compared with the model group, the body weight decreased; Bacteroidetes, Actinobacteria, Bacteroides and Clostridium decreased. Firmicutes, DeltaProteobacteria and Lactobacillus increased; fasting blood glucose and serum TNF-α decreased (P<0.01), and IL-10 increased (P<0.01), contents of acetic acid, propionic acid and n-butyric acid increased in Wumeiwan high-dose, medium-dose and low-dose groups and metformin group (P<0.05, P<0.01).Conclusion:Wumeiwan may prevent and treat T2DM by regulating intestinal flora, improving inflammatory response, increasing SCFAs content and reducing blood glucose.Keywords：Wumeiwan;type 2 diabetes;intestinal flora;inflammatory factors;short-chain fatty acids- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Classic Prescriptions
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Effect of Shenteng Sanhuang Decoction and Gegen Qinlian Tang on Diabetic Kidney Disease Rats
Abstract：Objective:To explore the protective mechanism of Shenteng Sanhuang decoction and Gegen Qinlian Tang on diabetic nephropathy (DN).Method:Rat DN models were duplicated with unilateral nephrectomy combined with streptozotocin (STZ). The rats were randomly divided into blank group, model group, irbesartan group and traditional Chinese medicine group. After 8 weeks of administration of corresponding drugs, the body weight, blood sugar, blood urea nitrogen (BUN), 24-hour urine volume (24 h U-vol), 24-hour urinary protein (24 h U-pro), serum creatinine (SCr), kidney weight/body weight (KW/BW) mass index of rats in each group were measured. The kidney tissues of rats in each group were homogenized, and supernatant was taken. Expressions of Smad ubiquitination regulatory factor 2 (Smurf2), matrix metalloprotein-9 (MMP-9), TGF-β1 and malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) were detected by Western blot or special test kit.Result:Compared with the blank group, the biochemical indicators, body weight, KW/BW, blood sugar, BUN, 24 h U-vol, 24 h U-pro, SCr and MDA were significantly higher or increased(P<0.05), while SOD and CAT were significantly decreased in the model group(P<0.05). Western blot showed that the expression of Smurf2 and TGF-β1 was high, while the expression of MMP-9 was low(P<0.05). Compared with the model group, the biochemical indicators of irbesartan group and traditional Chinese medicine group improved significantly(P<0.05), KW/BW were reduced, and blood sugar, BUN, 24 h U-vol, 24 h U-pro, SCr and MDA were significantly decreased (P<0.05), SOD and CAT was obviously increased (P<0.05), expressions of Smurf2 and TGF-β1 were decreased significantly(P<0.05), and expression of MMP-9 was increased significantly(P<0.05).Conclusion:Shenteng Sanhuang decoction and Gegen Qinlian Tang can effectively improve many biochemical indexes of rat DN models, and improve renal function. Its mechanism is closely related to reducing the expressions of Smurf2 and TGF-β1, and enhancing the expression of MMP-9.Keywords：diabetic nephropathy;Shenteng Sanhuang decoction;Gegen Qinlian Tang;renal function- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Toxicity and Mechanism of Polygoni Multiflori Radix Alcohol Extract on L02 Cells
Abstract：Objective:To study the toxic effect of Polygoni Multiflori Radix alcohol extract (PME) on L02 cells and the mechanism of ROS inducing apoptosis via mitochondria pathway, so as to provide a basis for the rational and safe administration of Polygoni Multiflori Radix in clinic.Method:The 4, 5-dimethly-2-thiazolyl-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay was used to detect the cell viability of PME at different concentrations (5, 10, 20 g·L-1). Nuclear morphology was observed by Hoechst 33342 staining. The apoptosis rate of cells was detected by Annexin V-FITC/PI. The release rate of lactate dehydrogenase (LDH), the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in the cells were detected by kit instruction. The changes of mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) were detected by flow cytometry. The relative protein expression levels of B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), cysteinyl aspartate proteinase-9(proCaspase-9) and cysteinyl aspartate proteinase-3 (proCaspase-3) in the PME-administered group were detected by Western blot.Result:After treatment with PME at the concentration of 5, 10, 20 g·L-1, the survival rate of L02 cells were decreased in a concentration and time-depended manner. After treatment with PME for L02 cells, nucleus shrinkage, fragmentation and chromatin condensation were observed under fluorescence after Hoechst 33342 staining. Annexin V-FITC/PI double staining showed a upward cell apoptosis rate in PME 20 g·L-1 group. Compared with the normal control group, the release rate of LDH was significantly increased (P<0.01), the intracellular ROS level was significantly increased (P<0.01), and the SOD activity was significantly decreased (P<0.01), while the MMP rate was significantly decreased in PME 5, 10, 20 g·L-1 groups (P<0.05). With the increase in the concentration of PME, proCaspase-3, proCaspase-9, Bcl-2 protein showed a significantly downward trend in PME 10, 20 g·L-1 groups (P<0.01), while the expression of Bax protein was significantly up-regulated in PME 20 g·L-1 group (P<0.05).Conclusion:The study illustrated that PME have toxic effects on L02 cells, which may destroy the structure of hepatocytes to a certain extent, promote ROS levels, induce oxidative stress, activate the mitochondrial pathway, and then activate apoptosis-related proteins to cause cells damage. It is suggested that ROS-mediated mitochondrial pathway was involved in PME-induced apoptosis.Keywords：Polygoni Multiflori Radix;hepatotoxicity;apoptosis;reactive oxygen species;mitochondrial pathway- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Naoxintong Inhibits LPS Induced BV2 Cell Pyroptosis by NLRP3/Caspase-1 Signaling Pathway
Abstract：Objective:To explore the effect of Naoxintong ethanol extract (NXT) on pyroptosis of BV2 microglia cells induced by lipopolysaccharide (LPS), and to explain the mechanism of pyroptosis based on NOD like receptor thermoprotein domain 3 (NLRP3)/cysteine-proteinase-1 (Caspase-1) pathway.Method:BV2 cells was treated with different concentrations of NXT(2, 10, 50 mg·L-1) after induced by LPS(1 mg·L-1) in vitro. Real-time PCR was used to detect mRNA expression of pro-inflammatory cytokine such as interleukin 1 beta (IL-1β), tumor necrosis factor (TNF)-α, and NLRP3.Western bolt and immunofluorescence were used to observe the protein expression of NLRP3/Caspase-1 signaling pathway.Result:Compared with control group, after LPS(1 mg·L-1) stimulation, BV2 cells viability was decreased. The mRNA expression levels of IL-1β, TNF-α and NLRP3 were significantly elevated(P<0.01), the protein levels of NLRP3 and Caspase-1 p20/Caspase-1 were also increased. After given NXT(2, 10, 50 mg·L-1), BV2 cells viability reversed which induced by LPS. Compared with LPS group, the mRNA expression of IL-1β, TNF-α and NLRP3 reduced obviously with given 50 mg·L-1NXT (P<0.05, P<0.01), significantly inhibited NLRP3 high protein expression and Caspase-1 p20/Caspase-1 expression(P<0.01).Conclusion:NXT can inhibit LPS induced pyroptosis of BV2 cells and the mechanism may closely related to NLRP3/Caspase-1 signaling pathway.Keywords：Naoxintong;microglia;lipopolysaccharide;pyroptosis;inflammasome;Buyang Huanwu Tang- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Hepatoprotective Effect of Portulacae Herba on Carbon Tetrachloride Induced Acute Liver Injury in Mice
Abstract：Objective:To explore the effect and mechanism of Portulacae Herba protecting carbon tetrachloride (CCl4)-induced acute liver injury.Method:Sixty Kunming mice were randomly divided into normal group, model group, silybin group (200 mg·kg-1) and Portulacae Herba high, medium, low (2, 1, 0.5 g·kg-1) dose groups. After continuous intragastric administration for 5 days, mice in each group were intraperitoneally injected with 0.2% CCl4 peanut oil solution to establish acute liver injury model, except normal mice. After 23 hours of modeling, serum and liver tissue were collected. Fully automatic analysis of serum serum liver function indicators in mice. Liver tissues were taken for hematoxylin-eosin staining (HE) staining to observe liver pathological changes. RNA Sequencing (RNA-seq) was used to analyze differential genes and functional enrichment, real-time fluorescence quantification PCR(Real-time PCR) was used to verify the mRNA expression of cytochrome P450 family members(CYP)26A1, CYP2C37, CYP2C44, CYP2C50, CYP2C54.Result:Compared with normal group, the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), total bilirubin (TBIL), malondialdehyde (MDA) in model group were significantly increased (P<0.05), and the activities of triglyceride (TG) and superoxide dismutase (SOD) were significantly decreased (P<0.05). Compared with model group, Portulaca Herba significantly reduced ALT, AST, TBIL and MDA levels in mice with acute liver injury (P<0.05), significantly increased SOD activity (P<0.01), and decreased the degree of liver tissue damage in mice. Compared with normal group, the mRNA expressions of CYP2C44, CYP2C50 in mice with acute liver injury were significantly decreased (P<0.05). Compared with model group, the mRNA expressions of CYP26A1, CYP2C37, CYP2C44, CYP2C50 and CYP2C54 were significantly increased in all dose groups of Portulaca Herba (P<0.05, P<0.01).Conclusion:Portulacae Herba has significant protective effects on acute liver injury caused by CCl4, and its mechanism may be related to the regulation of cytochrome P450 related genes.Keywords：Portulacae Herba;hepatoprotective effect;CCl4 induced acute liver injury;RNA sequencing;cytochrome P450;mice- 14
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Pharmacology
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Effect of Modified Shoutaiwan on Recurrent Abortion Due to Prethrombotic Status and Effect on Coagulation Factors
Abstract：Objective:To observe the efficacy of modified Shoutaiwan on recurrent abortion due to prethrombotic status with kidney deficiency and blood stasis and investigate its effect on coagulation factors.Method:The 100 cases were randomly divided into control group and observation group, 50 cases in each group. Bushen Yangxue capsule + enoxaparin sodium were given in control group, while modified Shoutaiwan + enoxaparin sodium were given in observation group, with a treatment course of 60 d in both groups. The serum clotting factor activated protein S (PS), activated protein C (PC), platelet aggregation function (PAF), platelet aggregation rate (PagT), antithrombin (AT), lupus anticoagulant material positive positive predictive value (LA), anti cardiolipin antibody positive rate of positive predictive value (ACA), plasminogen activator inhibitor-1(PAI-1), tissue factor (TF), fibrinogen (Fig), fibrinogen (FIB), fibrinogen alpha granule membrane glycoprotein-140 (GMP-140), fibrinogen fragment (F1+ 2), tissue fibrinogen activation factor (t-PA), D-dimer (D-D), activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT) were observed. Clinical efficacy, pregnancy rate, traditional Chinese medicine(TCM) syndromes and adverse reactions were compared between the two groups.Result:4 cases withdrew from the study. The total effective rate in observation group was 97.9% (48/49), higher than 85.1% (40/47) in control group (χ2=5.713, P<0.05). The pregnancy rate in observation group was 89.8% (44/49), higher than 70.2% (33/47) in control group (χ2=6.351, P<0.05). The score of traditional Chinese medicine syndromes in observation group was lower than that in control group. The indexes of PS, PC, AT, TF, Fig, FPA, APTT and TT in observation group were significantly higher than those in control group (P<0.05), while PAF, PagT, LA, ACA, PAI-1, PAP, TAT, FIB, GMP-140, F1+ 2, t-PA, D-D and PT were lower than those in control group (P<0.05). The incidence of adverse reactions in observation group was 10.2% (5/49), lower than 35.4% (17/48) in control group (P<0.05).Conclusion:Modified Shoutaiwan can effectively improve the clinical symptoms of recurrent abortion due to prethrombotic status and improve the coagulation factors in patients with kidney deficiency and blood stasis.Keywords：modified Shoutaiwan;enoxaparin sodium;prethrombotic status;recurrent abortion;kidney deficiency and blood stasis;clotting factor- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect and Mechanism of Modified Si Junzitang Combined with Heat-sensitive Moxibustion on IL-17, IL-22, IL-1
α and Cys-C in Serum and Exhale Breath Condensate of Patients with Chronic Obstructive Pulmonary Disease at Stable StageAbstract：Objective:To observe the effect and mechanism of modified Si Junzitang combined with heat-sensitive moxibustion on interleukin-17(IL-17), interleukin-22(IL-22), interleukin-1α(IL-1α) and serum cystatin C(Cys-C )in serum and exhale breath condensate(EBC) of patients with chronic obstructive pulmonary disease at stable stage(COPD, Qi deficiency of lung and spleen).Method:Totally 120 cases of COPD(Qi deficiency of lung and spleen) treated in pulmonary department and thermal moxibustion department of Affiliated Hospital of Jiangxi University of traditional Chinese medicine from January 2019 to June 2019 were included and randomly divided into traditional Chinese medicine group, heat-sensitive moxibustion group and control group. The patients in traditional Chinese medicine group were treated with Si Junzitang, the patients in heat-sensitive Moxibustion group were treated with heat-sensitive moxibustion in addition to traditional Chinese medicine group, and the patients in control group were treated with placebo. All of the 3 groups were treated with oxygen and bronchodilator according to the guidelines. All groups received 3 consecutive courses of treatment, 20 days per course. After 3 courses of treatment, the clinical efficacy of the three groups, the forced expiratory volume in one second (FEV1), the forced expiratory volume in the estimated value in one second (FEV1%), the forced vital capacity (FVC), and IL-17, IL-22, IL-1α in serum and exhale breath condensate (EBC) were measured.Result:There were no statistically significant difference in general clinical data, lung function levels (FEV1, FEV1%, FVC), serum and EBC levels of IL-17, IL-22, IL-1α and Cys-C in the first three groups. The total clinical effective rate of traditional Chinese medicine group was better than the control group (P<0.05), the heat-sensitive moxibustion group was better than the traditional Chinese medicine group (P<0.05) and significantly better than the control group (P<0.01). Compared with the patients before treatment, the level of lung function was improved, while IL-17, IL-22, IL-1α and Cys-C in serum and EBC were reduced(P<0.05). The traditional Chinese medicine group was superior to that in the control group (P<0.05), the heat-sensitive moxibustion group was superior to that in the traditional Chinese medicine group (P<0.05) and significantly superior to that in the control group (P<0.01).Conclusion:Modified Si Junzitang combined with heat-sensitive moxibustion has an anti-inflammatory effect on COPD by stimulating bullishness of human body, improving body immunity, inhibiting inflammatory cytokines, reducing levels of inflammation cytokines IL-17, IL-22, IL-1α, and chronic inflammation markers serum Cys-C and inflammatory reaction, increasing the lung capacity, improving ventilation function and pulmonary function, so as to effectively relieve chest tightness asthma and other symptoms in COPD patients, and improve the clinical efficacy.Keywords：chronic obstructive pulmonary disease (COPD);stable period;Si Junzitang;heat-sensitive moxibustion;exhale breath condensate (EBC);inflammatory cytokines;serum cystatin C(Cys-C);clinical observation- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Clinical Efficacy of Magui Wenjingtang Combined with Shoulder Joint Loosening Operation in Treating Periarthritis of Shoulder with Syndrome of Blood Deficiency and Cold Coagulation
Abstract：Objective:To explore the clinical efficacy of Magui Wenjingtang combined with joint mobilization in the treatment of periarthritis of the shoulder and the effect on hemodynamics and serum pain media.Method:A total of 120 patients with scapulohumeral periarthritis were enrolled in Tianjin Ninghe Hospital from July 2016 to July 2018.All cases were divided into control group and observation group, with 60 patients in each group. The control group received shoulder joint loosening operation (qd), and celecoxib capsules for oral administration. The observation group was treated with Magui Wenjingtang in addition to the shoulder joint loosening operation, 1 dose/d, 2 times/d. After 4 weeks of continuous treatment, the subjects were compared for the subjective pain level McGill Pain Questionnaire (SF-MPQ) score, shoulder mobility hemodynamic changes, and serum prostaglandin E2 (PGE2), substance P (SP), serotonin (5-HT) level and clinical efficacy.Result:Compared with before treatment, visual pain score (VAS), current pain status (PPI), and pain rating index (PRI) scores of the two groups were significantly lower after 4 weeks of treatment, and those of the observation group was significantly lower than the control group (P<0.01). The range of shoulder joint activity was significantly higher than that of the control group (P<0.01), erythrocyte sedimentation rate (ESR), fibrinogen (Fb), whole blood viscosity and serum were compared between the two groups after 4 weeks of treatment and before treatment. The levels of PGE2, SP and 5-HT were significantly lower (P<0.01), and the observation group was significantly better than the control group (P<0.01). The total effective rate of the observation group was 96.61%(57/59), which was significantly higher than 82.76% (48/58) of the control group(χ2=4.685, P<0.05).Conclusion:Magui Wenjingtang combined with shoulder joint loosening operation in treating periarthritis of shoulder with syndrome of blood deficiency and cold coagulation can effectively improve the pain and shoulder joint mobility, increase the curative effect, and improve the hemodynamics, and down-regulatie pain mediator of PGE2, SP, 5-HT, which may be related to the curative effect.Keywords：Magui Wenjingtang;periarthritis of shoulder;joint loosening operation;syndrome of blood deficiency and cold coagulation- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Clinical Efficacy of Modified Weijingtang and Maxing Shigan Tang Combined with Half-dose Hormones on Refractory Mycoplcasma Pneumoniae Pneumonia in Children Caused by Toxic Heat Closing Lung
Abstract：Objective:To observe the clinical efficacy of modified Weijingtang and Maxing Shigan Tang combined with half-dose hormones in the treatment of mycoplcasma pneumoniae pneumonia in children caused by toxic heat closing lung.Method:Totally 120 cases were randomly divided into control group and observation group, with 60 cases in each group. In addition to the basic therapy, control group was given Lianhua Qingwen granules + methanilone (10 mg·kg-1, 2 times/day), while observation group was given modified Weijingtang and Maxing Shigan Tang + methanilone (10 mg·kg-1, 1 time/day) for 14 days. Lung function indicators [maximal voluntary ventilation (MVV), time of peak tidal expiratory flow (TPTEF), forced expiratory volume in one second (FEV1), volume peak time expiratory flow (VPTEF)], plasma myocardial enzyme profile [creatine kinase (CK), creatine kinase isoenzyme (CKMB), lactate dehydrogenase (LDH), hydroxybutyric acid dehydrogenase (HBDH)], immune function [immunoglobulin G (IgG), immunoglobulin M (IgM), erythrocyte immune complex (RBC-ICR), erythrocyte C3b receptor (RBC-C3bR)], inflammatory factors [tumor necrosis factor-alpha (TNF-α), gamma-interferon (IFN-γ), interleukin-13 (IL-13), interleukin-17A (IL-17A)], clinical efficacy and adverse reactions were observed before and after treatment.Result:Four cases fell off during the study period. The total effective rate in observation group was 96.6%(57/59), which was higher than that in control group 84.2%(48/57, P<0.05). Compared with control group after treatment, lung function, RBC-C3bR and IL-13 were increased in observation group (P<0.05), while myocardial enzyme spectrum, IgG, IgM, RBC-ICR, TNF-α, IFN-γ and IL-17A were decreased in observation group (P<0.05). The incidence of adverse reactions in observation group was, lower than that in control group(P<0.05).Conclusion:Modified Weijingtang and Maxing Shigan Tang combined with half-dose hormones can improve lung function, myocardial enzyme profile, immune function and inflammatory factor levels of refractory mycoplasma pneumoniae in children caused by toxic heat closing lung, with a incidence of adverse reactions.Keywords：Weijingtang and Maxing Shigan Tang;half-dose hormones;refractory mycoplcasma pneumoniae pneumonia in children;toxic heat closing lung syndrome- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Protective Effect of Modified Qiju Dihuang Pills on Renal Function of Patients with Early Renal Impairment
Abstract：Objective:To observe the efficacy of modified Qiju Dihuang pills in protecting renal function of patients with early renal impairment with syndrome of Yin deficiency and Yang hyperactivity caused by hypertension and its effect in resisting inflammation and oxidative stress, and improving endothelial function.Method:Randomly divided into control group (59 cases) and observation group (60 cases) by random number table. Patients in control got valsartan capsules, 80 mg/time, 1 time/day. And patients whose blood pressure can't be controlled were added with nifedipine tablets, 10 mg/time, 1 time/day. In addition to the therapy of control group, patients in observation group were added with modified Qiju Dihuang pills, 1 dose/day. The control group took Qiju Dihuang pills.The courses of treatment were 16 weeks. And levels of cystatin C (CysC), urinary N-acetyl-β-glucosaminidase (NAG), β2 microglobulin (β2-MG), urinary microalbumin to creatinine ratio (UACR) and emodynamics of kidney were detected. Blood pressure, systolic pressure (SBP), diastolic pressure (DBP) and blood pressure renal artery resistance index (RI) and pulsatility index (PI) were recorded, and Yin deficiency and Yang hyperactivity were scored. Levels of nitric oxide (NO), endothelin-1 (ET-1), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), adiponectin (ADPN) and nuclear factor-kappa B (NF-κB) were detected.Result:The total effective rate in observation group was 91.67%(55/60), which was higher than 76.27%(45/59) in control group (χ2=5.255, P<0.05). Levels of SBP and DBP were lower than those in control group (P<0.05). At the 16th week during the treatment, the compliance rate of blood pressure was 90.66%, which was higher than 84.13% in control group (χ2=127.65, P<0.01). And levels of CysC, β2-MG, NAG, UACR, RI, PI, ET-1, SOD, GSH-Px, IL-6, NF-κB and TNF-α were lower than those in control group (P<0.01), while levels of NO, SOD and GSH-Px were higher than those in control group (P<0.01).Conclusion:In addition to the therapy for controlling blood pressure with routine western medicine, modified Qiju Dihuang pills can reduce level of the blood pressure, and control blood pressure with the standard, with anti-inflammatory and anti-oxidative stress effects. It can also improve the vasomotor function and the flow of kidney, protect the kidney function, and reduce the degree of injury, so as to delay the process of kidney damage and improve the prognosis.Keywords：hypertension;early renal impairment;modified Qiju Dihuang pills;Yin deficiency and Yang hyperactivity;inflammatory response;oxidative stress;endothelial function;renal hemodynamics- 6
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Wenyang Huashi Prescription on Inflammatory Factors and Immune Function of Patients After Allergic Fungal Rhinosinusitis
Abstract：Objective:To assess the efficacy of Wenyang Huashi prescription on rehabilitation of patients after allergic fungal rhinosinusitis (AFRS) operation, and its effect on inflammatory factors and immune function.Method:One hundred and forty-four patients were randomly divided into control group and observation group by random number table, with 72 cases in each group. Patients in control group was treated with Messerklinger to remove lesions completely, and got anti-infective therapy for three days, prednisone acetate tablets, 10 mg/time, 2 times/days, fluticasone propionate inhaled aerosol for a continued 4 weeks, 2 sprays/time, 1 time/day, itraconazole capsules at the first, fifth and ninth week after treatment (one course of treatment was 7 days), 0.2 g/time, 1 time/day. In addition to the therapy of meloxicam tablets, patients in observation group were also given local rinsing and oral administration of Wenyang Huashi prescription. The courses of treatment of both groups were 12 weeks. Before the operation and at the fourth and 12th week after the operation, subjective conditions were assessed. And before the operation and at the 12th, 24th and 48th week after the operation, Lund-Kennedy detected by endoscopic sinus surgery and nasal and paranasal sinus outcomes test-20 (SNOT-20) were scored. And the relapse was recorded during 48 weeks after the operation. And levels of tumor necrosis factor-alpha (TNF-alpha), interleukin-1bet (IL-1bet), nuclear factor kappa B (NF-kappa B), intercellular adhesion molecule in peripheral blood-1 (ICAM-1), T lymphocyte subsets (CD3+, CD4+, CD8+ and CD4+ /CD8+) were detected.Result:By rank sum test, the clinical efficacy in observation group was better than that in control group (Z=2.016, P<0.05). At the fourth and 12th week after the operation, scores of nasal obstruction, dizziness, runny nose and dysosmia in observation group were lower than those in control group (P<0.01). And scores of lund-kennedy and SNOT-20 were lower than those in control group (P<0.01). And levels of TNF-α, IL-1β, NF-κB and ICAM-1 were lower than those in control group (P<0.01). After treatment, levels of CD3+, CD4+ /CD8+ and CD4+ were higher than those in control group, and level of CD8+ was lower than that in control group (P<0.05). During the one-year follow-up, the relapse rate in observation group was 7.81%(5/64), which was lower than 22.58%(14/62) in control group (χ2=5.363, P<0.05).Conclusion:Wenyang Huashi prescription can improve short-term and long-term symptoms, improve the comprehensive curative effect, the quality of life and the immune function of the body, alleviate inflammatory injury, reduce the recurrence rate and improve the prognosis, and so is worth further study and use.Keywords：allergic fungal rhinosinusitis;Wenyang Huashi prescription;Mahuangfuzi Xixin Tang;Xinyi san;quality of life;recurrence;immune function;inflammatory factors- 6
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Clinic
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Effect of Polyethylene Glycol 400 on Bile Excretion of Baicalin and Its Main Metabolite
Abstract：Objective:To investigate the effect of polyethylene glycol 400 (PEG400) on rat bile excretion of baicalin and its main metabolite [baicalein 6-O-β-D-glucuronide (B6G)], and to analyze its mechanism of action.Method:Rats were randomly divided into baicalin+ water group and baicalin+ PEG400 group, the anesthesia was induced by intraperitoneal injection of 10% chloral hydrate (dose of 4 mL·kg-1) to prepare a rat bile duct intubation model. After the rats were fully awake, rats were given baicalin aqueous solution and baicalin PEG400 solution with dose of 168 mg·kg-1 for baicalin, respectively. And bile was collected from 0 h to 12 h after administration. UPLC-MS/MS was used to determine the concentration of drug excreted through bile at different time periods. Thermo Hypersil GOLD C18 column was used with acetonitrile (A)-0.1% formic acid solution (B) as the mobile phase for gradient elution (0-9 min, 90%-27%B; 9-10 min, 27%-90%B; 10-12 min, 90%B), the flow rate was 0.3 mL·min-1, the column temperature was 30 ℃, the injection volume was 5 μL. The mass spectra were obtained in positive ion mode with electrospray ionization (ESI). The effects of PEG400 on the activities and expressions in rat liver of uridine diphosphate glucuronyltransferase (UGT) 1A8 and UGT1A9 were studied in vitro incubation assay and enzyme linked immunosorbent assay (ELISA).Result:Compared with the baicalin+ water group, in the baicalin+ PEG400 group, the bile cumulative excretions of baicalin and B6G increased by 1.8 times and 2.1 times within 12 h, respectively. PEG400 increased the enzyme activities of UGT1A8 and UGT1A9 by 2.0 times and 1.5 times, and their concentrations in liver were increased by 2.2 times and 1.3 times, respectively.Conclusion:PEG400 can significantly increase the bile excretion of baicalin and its main metabolite B6G by enhancing the activities and expressions of UGT1A8 and UGT1A9, and its promoting effect on bile excretion of B6G is greater than that of baicalin, which provides a basis for the rational clinical application of PEG400 and the design of new dosage forms of flavonoids such as baicalin.Keywords：baicalin;baicalein 6-O-β-D-glucuronide;polyethylene glycol 400 (PEG400);bile;uridine diphosphate glucuronyltransferase (UGT) 1A8;UGT1A9;ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS)- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Drug Metabolism
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Simultaneous Determination of Six Flavonoids in Astragali Radix Directional Processed with Four Enzymes by UPLC
Abstract：Objective:To establish an UPLC method for the simultaneous determination of 6 flavonoids, and to research for the effect of Astragali Radix directional processed with four enzymes (complex enzyme, plant cellulase, snail enzyme, and β-glucosidase) on the contents of flavonoid glycosides and their aglycones in this herb.Method:Chromatographic separation was carried out on ACQUITY UPLC HSS T3 column (2.1 mm×100 mm, 1.8 μm) with the mobile phase of 0.1 mol·L-1 formic acid solution-0.1 mol·L-1 formic acid acetonitrile solution for gradient elution. The detection wavelength was set at 260 nm, the flow rate was 0.3 mL·min-1, the column temperature was 30 ℃, and the injection volume was 2 μL.Result:Calycosin-glucoside, calycosin, ononin, formononetin, 9, 10-dimethoxy-pterocarpan-3-O-β-D-glucoside and 3-hydroxy-9, 10-dimethoxy-pterocarpan showed good linear relationships within their own ranges (R2≥0.998 5), the relative standard deviations (RSDs) of precision, stability and repeatability were all <5.0%, and the average recovery was 97.62%-101.13% with RSDs of 1.4%-2.7%. In 0.5 g·L-1 level of enzyme solution, the contents of calycosin-glucoside, calycosin, ononin, formononetin, 9, 10-dimethoxy-pterocarpan-3-O-β-D-glucoside and 3-hydroxy-9, 10-dimethoxy-pterocarpan in Astragali Radix processed with complex enzyme were 0.082 0, 0.335 9, 0.055 9, 0.104 9, 0.015 0, 0.009 7 mg·g-1, the contents of them in Astragali Radix processed with plant cellulase were 0.105 7, 0.364 2, 0.070 2, 0.117 4, 0.020 8, 0.012 5 mg·g-1, their contents in Astragali Radix processed with snail enzyme were 0.031 4, 0.510 0, 0.043 5, 0.210 9, 0.013 0, 0.013 0 mg·g-1, and their contents in Astragali Radix processed with β-glucosidase were 0.085 3, 0.312 4, 0.061 5, 0.110 8, 0.005 8, 0.009 6 mg·g-1, respectively.Conclusion:After the processing of Astragali Radix by four enzymes, in addition to 9, 10-dimethoxy-pterocarpan-3-O-β-D-glucoside, the contents of calycosin-glucoside and ononin are reduced, but the contents of their three corresponding aglycones are significantly increased. The established method is simple, accurate and reproducible, and is suitable for the simultaneous determination of 6 flavonoids in Astragali Radix, which can provide a reference for this herb directional processed with enzymes.Keywords：Astragali Radix;ultra high performance liquid chromatography (UPLC);flavonoids;determination;directional processing;processing conversion;snail enzyme- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Analysis on Content Change and Transformation Rule of Main Characteristic Components in Stir-frying Process of Glycyrrhizae Radix et Rhizoma
Abstract：Objective:To study on the content change and transformation rule of eight main characteristic components in stir-frying process of Glycyrrhizae Radix et Rhizoma.Method:The contents of liquiritin apioside, liquiritin, isoliquiritin apioside, isoliquiritin, liquiritigenin, isoliquiritigenin, glycyrrhizic acid and glycyrrhetinic acid in Glycyrrhizae Radix et Rhizoma were determined simultaneously by HPLC. The chromatographic conditions were Waters Symmetry® C18 column (4.6 mm×250 mm, 5 μm), and the mobile phase of acetonitrile (A)-0.05%phosphoric acid solution (B) for gradient elution (0-9 min, 19%-25%A; 9-18 min, 25%-34%A; 18-38 min, 34%-51%A; 38-58 min, 51%-89%A), the flow rate of 1 mL·min-1, the detection wavelengths at 320 nm (0-16 min), 276 nm (16-25 min), 370 nm (25-28 min), 254 nm (28-58 min), the injection volume of 10 μL and the column temperature at 30 ℃.Result:After stir-frying, the total content of three components with dihydroflavone as mother nucleus was decreased, while the total content of three components with chalcone as mother nucleus showed an upward trend, the content change of glycyrrhizic acid was not obvious, but glycyrrhetinic acid content showed a slight upward trend. When the monomer heating temperature reached 130 ℃, dihydroflavones and chalcones could be isomerized with each other, and with the increase of temperature, the isomerization became more obvious. When the heating temperature rose to 180 ℃ (isoliquiritin apioside was 130 ℃), in addition to the isomerization, the glucosidic bond of flavonoid glycosides began to break and gradually transformed into the corresponding secondary glycosides or aglycones. Glucosidic bond of glycyrrhizic acid could also be broken to form glycyrrhetinic acid, which was detected at 150 ℃.Conclusion:The change of chemical composition is complex during stir-frying process of Glycyrrhizae Radix et Rhizoma, in addition to the isomerization and glucosidic bonds breaking observed in this experiment, there may be other complex reactions. The content of one compound in the herb is affected by many factors during its processing, such as the time and temperature of frying, the stability of the compound itself and so on.Keywords：Glycyrrhizae Radix et Rhizoma;stir-frying;content changes;high temperature;monomer component;flavonoids;triterpenes- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Rapid Identification on Chemical Components in Dalbergiae Odoriferae Lignum by UPLC-Q-TOF-MS/MS
Abstract：Objective:Dalbergiae Odoriferae Lignum is a rare traditional Chinese medicine material in China. However, there are many varieties of various sources and different qualities in the market at present. In order to further define the pharmacodynamic substance basis, electrospray time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) was used to rapidly analyze chemical constituents of methanol extract of Dalbergiae Odoriferae Lignum.Method:Chromatographic separation was performed on an UPLC RRHD SB-C18(3.0 mm×100 mm, 1.8 μm)for gradient elution, with mixtures of acetonitrile and 0.1%formic acid-water as mobile phases at a flow rate of 0.3 mL·min-1. The column temperature was maintained at 40 ℃. The data was collected in a negative ion mode with electro-spray ionization source(ESI).Result:According to molecular ion peaks and MS2 mass spectrometry characteristic fragment ions, Mass Bank databases, as well as the mass spectrometry information of reference substances and relevant literatures, a total of 83 constituents were identified, including 18 flavones, 31 isoflavones, 10 neoflavonoids, 9 isoflavanones, 7 other flavonoids and 8 other components.Conclusion:UPLC-Q-TOF-MS/MS can quickly, accurately and comprehensively identify chemical constituents in methanol extract of Dalbergiae Odoriferae Lignum, and isoflavones, flavones, neoflavonoids and isoflavanones are the main chemical constituents, which laid a foundation for the basic research of medicinal substances of Dalbergiae Odoriferae Lignum, and provided theoretical basis and technical support for the improvement of quality standards of Dalbergiae Odoriferae Lignum.Keywords：Dalbergiae Odoriferae Lignum;Dalbergia;UPLC-Q-TOF-MS/MS;chemical constituents;flavones;isoflavones;neoflavonoids- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Strong Light Stress on Growth, Physiological and Biochemical and Gene Expression of Key Enzymes in
Atractylodes lancea Abstract：Objective:To explore the effect of strong light stress on the growth, physiological and biochemical and key enzyme gene expression of the Atractylodes lancea, in order to provide the scientific basis for the standardized cultivation of the A. lancea.Method:The two-year-old A. lancea seedlings were taken as experimental materials. Poplar forest (light transmittance between 18.26%-36.04%) was taken as control group(ck). Different density shading networks were used to simulate different degrees of high light stress (51.10%, 80.73%, 100%) in late July. The growth state of A. lancea was observed. On the 0th, 5th, 10th, 15th, 20th days, the physiological and biochemical indexes of malondialdehyde (MDA) content, cell membrane permeability, proline (Pro) content, antioxidant enzyme activity and chlorophyll content in the leaves of A. lancea were measured. The relative expression levels of 3-hydroxy-3-methylglutarate monoacyl coenzyme A reductase (3-hydroxy-3-methylglutaryl coenzyme A, HMGR) and farnesyl pyrophosphate synthase gene (farnesyl pyrophosphate synthase, FPPS) in leaves of A. lancea under intense light stress were determined by real-time fluorescence quantitative PCR(Real-time PCR).Result:After strong light stress, the color of the leaves of A. lancea changed from dark green to light green and yellowish green, and the burn of leaves became more and more serious. The contents of MDA, conductivity and Pro showed an upward trend with the increase of transmittance. Peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT) tended to increase first and then decrease. The chlorophyll content decreased with the increase of light transmittance. The relative expression of HMGR in leaves of A. lancea decreased with the increase of light transmittance, while FPPS increased first and then decreased.Conclusion:The results showed that A. lanceaa could alleviate the inhibition of strong light stress by increasing the activity of antioxidant enzymes and regulating the content of osmotic pressure under certain strong light stress. Excessively strong intensity light stress leads to disequilibrium of metabolic mechanism of A. lancea, and seriously inhibits the plant growth.Keywords：Atractylodes lancea;strong light stress;3-hydroxy-3-methylglutaryl CoA reductase;farnesyl pyrophosphate synthase gene- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Establishment of a New Evaluation Method for Identification of Drug Resistance of
Staphylococcus aureus Based on Rapid Identification System of MicroorganismsAbstract：Objective:To explore the feasibility of the rapid identification system(MALDI-Biotyper System) of microorganisms for rapid identification of Pseudomonas aeruginosa and clinical isolation of Staphylococcus aureus.Method:Identification quality control and clinical isolation were conducted for drug resistance of S. aureus by microbial rapid identification system and broth dilution method. The scores of microbial rapid identification system were compared with the MIC value of broth dilution method. The drug resistance of P. aeruginosa was simultaneously identified to determine the accuracy and applicability of the rapid identification system of microorganisms.Result:The scores of the microbial rapid identification system showed that the score of sensitive quality control strain S. aureus was higher than 2.000, and the that of resistant strain of methicillin-resistant S. aureus(methicillin-resistant S. aureus, MRSA)was between 1.700 and 2.000.The score of clinically isolated S. aureus was between 1.700 and 2.000, which suggested the drug resistance and was consistent with the MIC value of the broth dilution method. At the same time, the systemic identification value of the P. aeruginosa, which is independent of the quality control sensitive strain, was greater than 2.000, showing sensitivity and it was a sensitive strain itself, which was consistent with the results.Conclusion:The microbial rapid identification system scoring method can be used for the rapid identification of the drug resistance of S. aureus and P. aeruginosa.Keywords：Staphylococcus aureus;Pseudomonas aeruginosa;methicillin-resistant Staphylococcus aureus;microdilution method;MALDI Biotyper System- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Quality Control of Salvianolic Acids Reference Extract
Abstract：Objective:To establish the quality control method for multi-index content determination and fingerprint of salvianolic acids.Method:Agilent ZORBAX SB-C18 (4.6 mm×250 mm, 5μm) column was adopted, with 0.1%formic acid-water as mobile phase A and 0.1%formic acid-acetonitrile as mobile phase B for gradient elution (0-30 min, 20%-21.5%B; 30-35 min, 21.5%-25%B; 35-45 min, 25%-40%B; 45-50 min, 40%-95%B). The column temperature was set at 30 ℃, the flow rate was set at 1 mL·min-1, and the detection wavelength was set at 288 nm. Relative correction factors of caffeic acid, salvianolic acid E, rosmarinic acid, lithosperic acid, salvianolic acid B and salvianolic acid Y were determined by the concentration method. The content of each indicator component of the reference extract of salvianolic acid polyphenolic acid was determined and compared with the results of the monomer reference substance by the external standard method. At the same time, the fingerprint method was established. and the similarity evaluation was carried out on 10 batches of extracts.Result:Caffeic acid, salvianolic acid E, rosmarinic acid, lithospermic acid, salvianolic acid B, and salvianolic acid Y had a good linear relationship within the respective detection mass concentration ranges (r>0.999 9). The injection precision RSD was 0.1%-1.2%, the reproducible RSD was 1.2%-1.6%, and the recovery of the six components was 82.03%-98.68%. The stability of each component in the sample solution was good within 36 h. The relative correction factors for each indicator component were determined to be caffeic acid (2.92), salvianolic acid E (1.10), rosmarinic acid (1.61), lithosperic acid (1.07), salvianolic acid B (1.00), salvianolic acid Y (0.83). The effects of different methods, concentrations, instruments, columns, wavelengths were investigated, and the measured relative correction factors were found to be suitable. The results of the calibration factor method and the monomer standard reference substance method were less different. The HPLC fingerprints of the reference extract of salvianolic acids were established, and five common characteristic peaks were determined. The chromatographic peaks were confirmed according to the reference substance. The similarity of the fingerprints of the 10 batches of extracts was higher, and the quality difference was smaller.Conclusion:The multi-index content determination method and the fingerprint method established in this study are simple, rapid, accurate and reproducible, and can be used for quality control of Salviae miltiorrhizae Radix et Rhizoma polyphenolic acid reference extract.Keywords：Reference extract;Salviae Miltiorrhizae Radix et Rhizoma;relative correction factor;fingerprint;salvianolic acid B;quality control- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Chemical Constituents from Stems of
Zanthoxylum bungeanum Abstract：Objective:To isolate and identify the chemical constituents from the 95%, 75%ethanol extracts of the stems of Zanthoxylum bungeanum.Method:The 25 kg stems of Z. bungeanum were extracted with 95%, 75%ethanol for three times, and the combined filtrates were concentrated under vacuum to get the extracts. The 95%extracts were then extracted by petroleum ether, dichloromethane, ethyl acetate and n-butanol successively to obtain corresponding fractions. Such fractions and 75%extracts were isolated and purified by silicagel, Sephadex LH-20, ODS, preparation HPLC and recrystallization to obtain compounds. Their structures were identified by mass spectroscopy (MS), and nuclear magnetic resonance (NMR).Result:Sixteen compounds were isolated from the stems of Z. bungeanum and identified as dictamnine(1), decarine(2), zanthobungeanine(3), pseudocolumbamine(4), skimmianine(5), norchelerythrine(6), osthenol(7), dimethylfraxetin(8), 3, 4, 5-trimethoxycinnamylalcohol(9), asarinin(10), yangambin(11), syringaresinol(12), ashantin(13), bis(2-ethylhexyl) benzene-1, 2-dicarboxylate(14), 24-propylcholesterol(15), and sucrose(16).Conclusion:Compounds pseudocolumbamine(4), 3, 4, 5-trimethoxycinnamylalcohol(9), and 24-propylcholesterol(15)were isolated from the genus of Zanthoxylum for the first time and compounds dictamnine(1), osthenol(7), dimethylfraxetin(8), asarinin(10), yangambin(11), syringaresinol(12), ashantin(13), and bis(2-ethylhexyl) benzene-1, 2-dicarboxylate(14)were isolated from this plant for the first time.Keywords：Zanthoxylum bungeanum stems;alkaloid;coumarin;lignans;ethanol extracts;pseudocolumbamine;3, 4, 5-trimethoxycinnamylalcohol;24-propylcholesterol- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Pre-column Derivatization RP-HPLC for Simultaneous Determination of 17 Amino Acids in Cynomorii Herba from Different Producing Areas and Its Multivariate Statistical Analysis
Abstract：Objective:To establish a pre-column derivatization reverse-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous determination of 17 amino acids in Cynomorii Herba from different producing areas and conduct a multivariate statistical analysis.Method:RP-HPLC with pre-column derivatization was employed, with phenyl isothiocyanate (PITC) as derivatization reagent. Separation was performed on a WondaSil C18-WR column (4.6 mm×150 mm, 5 μm), with 0.05 mol·L-1 sodium acetate solution (pH 6.5) as mobile phase A, and acetonitrile-methanol-water (3∶1∶1) as mobile phase B for gradient elution at a flow rate of 0.8 mL·min-1. The detective wave length was set at 254 nm, and the column temperature was maintained at 35 ℃. Principal component analysis (PCA) and systematic cluster analysis (HCA) models were established for multivariate statistical analysis and quality evaluation.Result:17 Kinds of amino acid were detected in Cynomorii Herba, 7 of which were essential amino acids. The 17 amino acids showed good linearity in respective concentration range, r = 0.999 0-0.999 9.The average recoveries were between 98.03%-103.89%with RSD<3.5%. The results of PCA and HCA were basically the same, and both methods can be used to clearly distinguish Cynomorii Herba from 12 municipal producing areas into 6 regions. PCA can be used to classify Cynomorii Herba according to different municipal or provincial production areas, and HCA can be used to classify it according to provincial production areas. It showed that the amino acid contents in Cynomorii Herba from different municipal and provincial producing areas had differences, and the content distribution showed obvious geographical clustering characteristics. PCA showed that Cynomorii Herba from Gansu province and Inner Mongolia had higher amino acid contents and better quality as compared with other producing areas.Conclusion:The established method can be used for content determination of 17 amino acids in Cynomorii Herba from different producing areas, and provide a reference for its comprehensive quality evaluation.Keywords：Cynomorii Herba;amino acid;PITC;pre-column derivatization;RP-HPLC;principal component analysis;systematic cluster analysis;quality evaluation- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Detection of Rutin in Sophorae Immaturus Flos with Electrochemical Sensor Assembled with Gold Nanoparticles/Nafion/Multi-wall Carbon Nanotube
Abstract：Objective:A new-type electrochemical sensor was developed by electrodepositing gold nanoparticle on the surface of glassy carbon electrode (GCE) modified by Nafion-multi-wall carbon nanotube (MWNTs) by the potentiostatic method, and used for the detection of rutin in practical samples.Method:The electrochemical properties of the sensor in potassium ferricyanide-potassium ferrocyanide system were studied by electrochemical impedance spectroscopy and cyclic voltammetric method, in order to explore the electrochemical performance of the sensor. The electrochemical behavior of rutin on the surface of the sensor was studied by the cyclic voltammetric method, and the factors affecting the performance of the sensor were optimized. The content of rutin was determined by differential pulse voltammetric methods.Result:The optimized conditions were -0.25 V of the deposition potential of gold nanoparticle, 20 s of the deposition time, pH 3.0, and 6 μL of 1 g·L-1 carbon nanotube. Under the conditions, a good linear relationship between reduction peak current and rutin concentration was obtained from 5.0 ×10-9 to 7.0 ×10-7 mol·L-1, and the detection limit was 3.6×10-9 mol·L-1.Conclusion:The developed sensor has a good electrical conductivity and stability, and is highly sensitive and simple for the detection of rutin, with a low detection limit. It has been successfully applied to the detection of rutin in Sophorae Immaturus Flos with the recoveries between 97.6%and 104.4%, which provides a new method for the determination of rutin and a new idea for the quality control of Chinese medicinal materials.Keywords：electrochemical sensor;multi-wall carbon nanotubes;gold nanoparticles;Nafion;rutin- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Pharmacy Fundamentals
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Active Components and Mechanism of Rhei Radix et Rhizoma on Renal Fibrosis Based on Network Pharmacology
Abstract：Objective:To explore the active components, potential targets and signaling pathways of Rhei Radix et Rhizoma in the treatment of renal fibrosis based on the network pharmacology method, and then to verify the target genes in vitro.Method:Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and Traditional Chinese Medicine Integrated Database (TCMID) were retrieved to obtain the main active ingredients of Rhei Radix et Rhizoma. The potential anti-renal fibrosis targets of Rhei Radix et Rhizoma were predicted by similarity ensemble approach (SEA), Swiss Institute of Bioinformatics (SIB) and GeneCards Database. Target protein-protein interaction (PPI) network was constructed by using String Version 10.5 database. David 6.8 software was used for gene ontology (GO) enrichment analysis and the Kyoto Encyclopedia of genes and genomes (KEGG) pathway enrichment analysis of the key targets. Cytoscape Version 3.6.0 software was used for visualized analysis of PPI network, active ingredient-key target network and the ingredient-target-signal pathway network. In combination with Malachards database, the signal pathways with high correlation with renal fibrosis were screened. Then, cell experiments were used for verification: HK-2 cells were selected to establish fibrosis model by transforming growth factor-β1 (TGF-β1) stimulation. The cells were treated with rhein for 48 hours. Western blot assay was used to detect the protein expression level of hypoxia inducible factor-1 α (HIF-1 α), vascular endothelial growth factor (VEGF), and platelet-derived growth factor receptor-α (PDGFR-α). Protein expression levels of E-cadherin and α smooth muscle actin (α-SMA) were detected by immunofluorescence. Apoptosis was detected with flow cytometry.Result:Totally 17 active ingredients of Rhei Radix et Rhizoma and 424 targets of anti-renal fibrosis effect were screened out, including five key targets: protein kinase B(Akt)1, mitogen activated protein kinases 3(MAPK3), epidermal growth factor receptor(EGFR), interleukin(IL)-6 and VEGFA in turn. The biological process of GO enrichment mainly involved signal transduction, cell proliferation and apoptotic process. The results of KEGG pathway enrichment showed that phosphatidylinositol 3-kinase(PI3K)/Akt, HIF-1, VEGF, and forkhead transcription factor (FoxO) pathways were related to the anti-renal fibrosis mechanism of Rhei Radix et Rhizoma. Results of the in vitro experiment proved that rhein could inhibit the expression of E-cadherin, α-SMA, HIF-1α, VEGF and PDGFR-α. In addition, rhein inhibited apoptosis induced by TGF-β1 in HK-2 cells. Part of the prediction results of network pharmacology were verified.Conclusion:This study reflects the multi-component, multi-target and multi-pathway mechanism characteristics of Rhei Radix et Rhizoma. The mechanisms of its anti-renal fibrosis effects may be related to inhibiting HIF-1 α / VEGF /PDGFR-α signaling pathway, apoptosis and epithelial-mesenchymal transition (EMT) of renal tubular epithelial cells.Keywords：Rhei Radix et Rhizoma;network pharmacology;renal fibrosis;target;mechanism of action- 13
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Visualization Analysis of Henoch-schonlein Purpura Treated by Traditional Chinese Medicine Based on Citespace
Abstract：Objective:To finding the main research contents, research frontier, author and institutional cooperation of traditional Chinese medicine(TCM) for treating henoch-schonlein purpura(HSP). Providing reference for the research and development of TCM for treating the disease.Method:Using Citespace to analyze 2 878 TCM articles related to HSP retrieved from CNKI, cluster analysis and burst analysis of literature keywords, co-occurring authors and institutional cooperation analysis.Result:Since 1995, the number of related literature was growing rapidly and had been stable at more than 100 per year after 2005.Cluster analysis showed 32 clusters, consisting of 396 nodes and 638 lines. The main clustering results include Children with allergic purpura, blood-activating and stasis-resolving drug, Henoch-Schonlein purpura nephritis, blood-cooling drugs, clinical observation, etc. Break analysis yielded 52 emergent words. It can be seen that TCM treatment of HSP is mainly based on cooling blood, followed by activating blood to eliminate stagnation and clearing heat. Commonly used drugs are Moutan Cortex, Paeoniae Radix Rubra, and Rehmanniae Radix, etc. Clinically, it pays more attention to the experience of famous doctors, research on Children with allergic purpura, etc.The author's cooperation network has obtained the maps of the three main cooperation teams with DING Ying, SUN Yi-qiu and HE Ping as the core. The Density of institutional cooperation network is 0.007 1.Conclusion:The main research contents of TCM for treating HSP include Children with allergic purpura, blood-activating and stasis-resolving drug, HSP nephritis, blood-cooling drugs, clinical observation, etc. Children with allergic purpura, experience from famous doctor, HSP nephritis and clinical efficacy is the foremost current research hotspot. A number of research teams have been formed that are relatively stable, but the institutional cooperation is scattered.Keywords：visualization;Citespace;henoch-schonlein purpura;traditional Chinese medicine;mapping knowledge domains- 20
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Data Mining
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Research and Practice of " Three Suitability" Theory in Processing of Traditional Chinese Medicine
Abstract：As an important part of clinical medication, the main function of processing of traditional Chinese medicine (TCM) is to reduce toxicity and increase efficiency. The key to the best clinical efficacy of TCM after processing lies in three aspects of moderation, adaptation and timeliness, namely " three suitability" theory. In recent years, scholars have done a lot of research under the guide of " three suitability" theory, fully explaining the scientific connotation of the theory, which greatly promoted the inheritance and innovation of TCM processing. In this paper, the basic connotation of " three suitability" theory was summarized, combining with a large number of modern research reports, the pharmacodynamics and composition changes of processed drugs were analyzed and discussed under the guide of " three suitability" theory. At the end of the paper, the authors proposed that we should not only systematize the traditional processing theories, but also use modern advanced technologies to explain the essence of traditional processing theories, and to innovate and develop new processing theories, promote the development of TCM processing discipline and the progress of TCM processing industry.Keywords：processing of traditional Chinese medicine;moderation;adaptation;timeliness;"three suitability" theory;essence of theory;inheritance and innovation- 13
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Correlation Analysis of " Toxin Damaging Brain Collaterals" from Glymphatic System
Abstract：The collaterals are branches of the meridians and vessels system, and have the roles of connecting upper-lower and interior-exterior portions of the body, the characteristics of two-way flow in supporting the operation of Qi and blood, and the functions of material exchange and metabolism. The brain is the intersection of the Yang meridians. Crisscross brain collaterals permeate Qi and blood to enrich the brain, and spread Yang Qi, in order to warm the brain-mind, and provide material basis and source power to the " brain governing mind" . Under pathological conditions, cerebral collaterals are blocked, and toxic pathogens are endogenous, resulting in " toxin damaging brain collaterals" . This theory is not only applied to the study of stroke pathogenesis, but also extended to other encephalopathy, such as dementia, which promoted the development of the theory of pathogenesis in traditional Chinese medicine. Recently, a " glymphatic system" was discovered in the brain, which is an exchange flow system of cerebrospinal fluid-brain interstitial fluid mediated by astrocyte. The glymphatic system transports nutrients and neuroactive substances, such as glucose, lipids, electrolytes and apolipoprotein E in the cerebrospinal fluid, to brain tissue, and also removes metabolic products (such as lactic acid), soluble proteins (such as β-amyloid protein and Tau protein) from the brain and foreign bodies, which are important liquid flow systems that maintain the homeostasis of the brain. The discovery of the glymphatic system provides a new perspective for the study of pathological mechanism of neurological diseases, and may become a new target for interventions in neurological diseases, such as cerebrovascular diseases and neurodegenerative diseases. As a widely distributed cerebral metabolic waste removal way and material delivery system, the lymphatic system may be the biological foundation of " brain collateral" disease, and a cross point of understanding on " toxin impairing brain collaterals" by Western and traditional Chinese medicine. The study based on the glymphatic system will give a more rational explanation on " toxic damage to brain collaterals" .Keywords：collaterals;toxin damaging brain collaterals;glymphatic system- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Theory Analysis on Absorption of Exogenous Substances in Traditional Chinese Medicine
Abstract：The safety of traditional Chinese medicine is affected by many factors, and the influence of exogenous harmful substances has been concerned and become a hot spot in recent years, especially heavy metals, pesticide residues and some other harmful substances. In order to explore the effects of non-soil and non-pesticide treatment on residues of these two harmful substances, the heavy metals and agricultural residues of Lilii Bulbus were detected, and the correlation of the data was analyzed. In this experiment, heavy metals and pesticide residues of Scrophulariae Radix were detected, and correlation analysis was conducted for their data. The mechanism of transport phase was interpreted with statistical moment similarity tool of total fingerprint by supramolecular chemistry theory. A large number of experimental data in this paper showed that heavy metals and pesticide contents in Lilii Bulbus basaltifolia had a positive correlation, which was closely related to supramolecular phenomena. Moreover, the similarity of fingerprints between Lilii Bulbus and Scrophulariae Radix suggested that Lilii Bulbus and Scrophulariae Radixa had a high selectivity in absorption of agricultural residues, which proved that the absorption of pesticides in Lilii Bulbus and Scrophulariae Radix from different habitats had the function of supramolecular imprinting template. It was considered that medicinal plant was a giant complex supramolecule with various levels of " imprinted template" . Heavy metals and agricultural residues were also involved in plant growth, forming an " imprinted template" for the formation of supramolecules between agricultural residues and heavy metals. After heavy metals and agricultural residues formed supramolecules, their liposolubility and permeability changed in varying degrees, and their transport in medicinal plants was promoted. Finally, the heavy metal supramolecules of pesticides were absorbed, distributed, aggregated and accumulated in plants. The purpose of the study was to reveal the mechanism of heavy metal and pesticide supramolecule transport, provide a new direction for the treatment of heavy metals and pesticide residues, and ensure the safety of traditional Chinese medicine.Keywords：heavy metal;agricultural residue;supramolecular theory;transport;Scrophulariae Radix;Lilii Bulbus- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Academic Discussion
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Herbal Textual Research of Setariae Fructus Germinatus
Abstract：Setariae Fructus Germinatus (Guya) is a commonly used traditional Chinese medicine, which has been used for thousands of years. In ancient and modern books and works, the name is often confused because of its complicated relationship with the origin. In order to clarify the name and source of Guya, the authors examined the name, origin and processing history of Guya through consulting ancient Chinese herbal books, modern Chinese medicine monographs, calendar edition of Chinese Pharmacopoeia and the processing standards of various provinces, and found that different regions in China used Guya according to their local habits, resulting in the foreign body of the same name of Guya, lacking a unified standard. It is suggested that changing the name of Guya to Suya is more practical, and is conducive to the quality standard research and clinical accurate application of Guya.Keywords：Setariae Fructus Germinatus;Suya;product name;origin;processing;herbal textual research;quality standard- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Herbal Textual Research and Modern Analysis of Pheretima
Abstract：Pheretima has a long history of medication, its original name was earthworm, and it was first recorded in Shennong Bencaojing, which was listed as inferior product. In the 2015 edition of Chinese Pharmacopoeia, two varieties of Pheretima were included. However, due to the variety of Pheretima, there are mixed non-pharmacopoeia collection of Pheretima family status. Based on the systematic review of ancient and modern literature, this paper conducts herbal textual research on Pheretima in terms of name, origin, distribution of origin, genuine production area, harvesting time, processing methods, etc. Based on the analysis of various ancient books and modern research documents of herbal medicine and their accompanying drawings, it is found that the Pheretima and white-necked Pheretima mentioned in ancient books are the general names of the genus Pheretima. The ancient people thought that white-necked Pheretima was a good medicinal material, which was the same as the opinion that Guangdilong was better in quality than Tudilong in modern research. In ancient and modern literature, the origin of earthworm is relatively consistent, but due to the change of environment, the output of wild Pheretima is reduced, and now the output of Pheretima is mainly artificial breeding. In ancient times, harvesting should be as far as possible in spring, summer and autumn. However, in modern times, the best harvest time is autumn. Different processing methods of earthworm in different ages and regions are different. Attention should be paid to following and inheriting the ancient processing methods, combining with modern research techniques, the quantitative standard of processing of Pheretima should be formulated, so that Pheretima medicinal materials can be applied comprehensively and effectively. This research provides the basis for the original source, resource development, correct use, genuine producing area and processing method determination of Pheretima.Keywords：Pheretima;herbal textual research;modern research;ancient and modern literature;origin;genuine producing area;processing- 16
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Herbal Textual Research
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Naphthoquinones from
Rhinacanthus nasutus and Their Pharmacological ActivitiesAbstract：Rhinacanthus nasutus is the shrubs of Acanthacesa of Rhinacanthus nasutus. It has been used as traditional Zhuang medicine for thousands of years, with antipyretic and alexipharmic, antibacterial and anti-inflammatory, antiasthmatic and expectorant effects. Modern pharmacological studies have shown that R. nasutus has been used for treatment of various diseases, including pneumonia, gastritis and hepatitis. In addition, naphthoquinone esters isolated from this plant exhibits antitumor, antiviral, antifungal, and immunomodulatory activities, with a high application value. Scholars at home and abroad have isolated a variety of compounds from R. nasutus, including 20 naphthoquinones, organic acids, lignans, alkaloids and other structural types of components. Among them, naphthoquinones have a variety of biological activities. Up to now, there are no systematic and comprehensive report on the chemical constituents and pharmacological effects of naphthoquinones. In this review, we summarized the chemical constituents of naphthoquinones with pharmacological activity, finding that R. nasutus and naphthoquinones have a great potential to be developed as a traditional Chinese medicine for tumors, diabetes, nonalcoholic fatty liver, which can provide an important scientific basis for defining the pharmacodynamic basis of the pharmacological activities and a reference for further research and comprehensive utilization of Zhuang medicine of Rhinacanthus nasutus.Keywords：Rhinacanthus nasutus;naphthoquinones;pharmacological activities;research progress- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Research Progress of Different Components of Traditional Chinese Medicine in Treatment of Blood Stasis Syndrome
Abstract：To a certain extent, put forward the concept of " component of traditional Chinese medicine (TCM)" simplifies the complexity of multi-component and multi-target of TCM, which provides a possibility for the clarification of the material basis of the efficacy of TCM, and also provides a new direction for promoting the modernization and industrialization of TCM, promots the high quality development of TCM. The correlation between prescription and disease syndrome has made rapid progress, both basic research and clinical application are fruitful. However, the correlation between components and disease syndrome still needs to be further studied. The syndrome of blood stasis is a common syndrome of TCM science, and it is more common in various diseases, especially cardiovascular and cerebrovascular diseases, kidney disease, diabetes and hyperlipoidemia. A large number of studies have shown that some specific components contained in TCM or TCM compound can improve the related indexes of patients or experimental animal model with blood stasis syndrome. It is manifested in reducing blood viscosity, inhibiting platelet activation and adhesion aggregation, changing erythrocyte deformability index, inhibiting thrombosis and so on. Blood stasis is not only the pathogenic factor of many diseases, but also the pathological product of many kinds of diseases, which involves a wide range of diseases. Therefore, this study will study the progress of different components of TCM in the prevention and treatment of blood stasis syndrome, focusing on saponins, flavonoids, organic acids, polysaccharides, alkaloids and other active components in improving hemorheological abnormalities, hypercoagulability, platelet activation and adhesion aggregation, thrombosis. Based on the thought of component-disease syndrome, this paper searches the relevant literature in recent 20 years, classifies and summarizes the achievements of different components in the prevention and treatment of blood stasis syndrome, and hopes to provide some ideas for the further study of the pharmacological action of TCM components, the study of compatibility of TCM components and the research of TCM components.Keywords：blood stasis syndrome;components of traditional Chinese medicine;research progress;promoting blood circulation and removing blood stasis;Saponins;flavonoids;organic acids;polysaccharides- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Traditional Chinese Medicine and Its Active Ingredients on Cardiovascular Disease Based on Nrf2/HO-1 Signaling Pathway
Abstract：Cardiovascular disease is the leading cause of death in China. Due to its great individual differences in genetic background, pathogenesis and disease development trend, the survival risk rate after standardized western medicine treatment under the guidance of the current guidelines remains high. Traditional Chinese medicine(TCM) has unique multiple-target, multiple-pathway and multiple-layer advantages, which can effectively make up for shortcomings of western medicine. Therefore, it has been widely used in the treatment of cardiovascular diseases. Oxidative stress is one of the important causes of cardiovascular diseases. Nuclear factor erythroid-2-related factor 2(Nrf2) is the central regulator of this reaction. When being activated, it can transfer to the nucleus and initiate signaling in the downstream pathway, thus playing an anti-oxidative stress role. As one of the most important endogenous protection systems in the body, the Nrf2/heme oxygenase-1 (Nrf2/HO-1) signaling pathway is the most classical approach for Nrf2 in playing roles. There have been certain achievements in studying and clarifying TCM by regulating this pathway to treat cardiovascular diseases using modern molecular biology and other methods. Based on this, this paper summarized the relationship between Nrf2/HO-1 signaling pathway and cardiovascular diseases, then concluded and analyzed the mechanism and pharmacological effects of TCM and its active ingredients in Nrf2/HO-1 signaling pathway on different cardiovascular diseases, involving active ingredients of TCM, TCM pairs active ingredients, TCM extracts and TCM formula. This paper provides a theoretical reference for the development and utilization of anti-cardiovascular drugs.Keywords：nuclear factor erythroid-2-related factor 2/heme oxygenase 1(Nrf2/HO-1);traditional Chinese medicine;active ingredients;cardiovascular disease- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Review
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