Abstract：Objective:To explore the effect of Tongxie Yaofang on p38 mitogen activated protease(p38 MAPK), mitogen and stress protein kinase 1(MSK1), cyclic adenosine effector response element binding protein(CREB)mRNA and protein expression in colon tissue of diarrhea type irritable bowel syndrome (D-IBS) rat model with liver depression and spleen deficiency(GYPX), and interleukin-1β(IL-1β), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), and the content of total superoxide dismutase (T-SOD) and malondialdehyde (MDA) in serum.Method:The 60 SPF Wistar rats were randomly divided into 6 groups, with 10 rats in each group. Except the normal group, rats in the other groups were given by gavage with folium sennae and chronic bondage to establish D-IBS with GYPX for 14 days. The low, medium, and high doses Tongxie Yaofang were administered to Tongxie Yaofang(2.25, 4.5, 9 g·kg-1)gavage respectively. The piveronium bromide group was given piveronium bromide tablets suspension(0.02 g·kg-1)gavage.The normal group group and model group were given the same volume normal saline for 21 days. After the last gavage for 18 hours, the heart blood was collected and the colon tissue was dissected. Real-time PCR was used to observe the expression of p38 MAPK, MSK1 and CREB mRNA in rat colon. Western blot was used to observe the expression of p38 MAPK, MSK1 and CREB protein. ELISA was used to observe the contents of IL-1β, IL-6 and TNF-α in colon. Hydroxylamine was used to observe the T-SOD level in serum, thiobarbituric acid(TBA)was used to observe the MDA content in serum. hematoxyl in-eosin(HE) staining was used to observe the morphological changes of colon tissues.Result:Compared with normal group, the expression of p38 MAPK, MSK1, CREB mRNA and the protein content of p38 MAPK, MSK1 and CREB in the colon tissue of model group rats increased significantly, while the content of IL-1β, IL-6 and TNF-α increased significantly(P<0.05). The level of serum T-SOD decreased significantly, and the content of MDA increased significantly(P<0.05). Compared with the model group, the medium and high dose group of Tongxie Yaofang significantly decreased the expression of p38 MAPK mRNA, content of p38 MAPK, CREB protein and IL-1β, IL-6, TNF-α in colon tissue(P<0.05). The level of serum T-SOD increased significantly, and the content of MDA decreased significantly(P<0.05). High dose group of Tongxie Yaofang can significantly decreased the expression of MSK1, CREB mRNA, content of MSK1 protein(P<0.05). Histopathological observation showed that no significant organic lesions were observed in the colonic morphology of each group of rats, which was consistent with the morphological characteristics of IBS.Conclusion:Tongxie Yaofang has a significant dose-effect relationship in the treatment of D-IBS rats with GYPX in a certain range, which may be related to its increases antioxidant stress and inhibit activation of p38 MAPK signaling pathway and reducing the level of downstream inflammatory factors.
Abstract：Objective:To study the anti-depressive effect of Qing' ewan in treating chronic unpredictable mild stress (CUMS) in rats, and the regulatory effect on estrogen receptor and estrogen receptor-related signaling pathways, in order to explore its anti-depressive mechanism.Method:The CUMS model was established. The experiment was divided into normal control group, model group, escitalopram oxalate group (positive control) and Qing' ewan groups (1.71, 5.13, 15.39 g·kg-1). After 4 weeks of modeling, rats were treated with corresponding drugs for 2 weeks. Behavioral evaluation [sucrose preference test (SPT), forced swimming test (FST), open field test (OFT)] was conducted to assess if the CUMS model was successful. Western blot was used to analyze the protein expression levels of estrogen receptor α (ERα), estrogen receptor β (ERβ), brain-derived neurotrophic factor (BDNF) and tyrosine kinase receptor B (TrkB).Result:Compared with the normal group, the sucrose consumption rate and the score of OFT in the model group decreased(P<0.05, P<0.01), the immobility time of FST prolonged significantly(P<0.01), and the protein expression levels of ERα, ERβ, BDNF and TrkB decreased(P<0.05, P<0.01). Compared with the model group, the behavioral performance of the treated group was improved, the sucrose consumption rate and the score of OFT increased(P<0.05, P<0.01), and the immobility time decreased(P<0.05). The protein expressions of ERα, ERβ, BDNF and TrkB in the treated group were significantly up-regulated(P<0.05, P<0.01), especially the middle-dose Qing' ewan group (5.13 g·kg-1).Conclusion:Qing' ewan can improve depression-like behavior in CUMS rats. Its mechanism may be related to the neuroprotective effect by up-regulating the expressions of ERα and ERβ and activating estrogen receptor-mediated ERβ/BDNF/TrkB pathways.
Abstract：Objective:To develop high performance liquid chromatography-diode array detector (HPLC-DAD) wavelength switching for simultaneously determining the contents of inosine, loganic acid, chlorogenic acid, amygdalin, hydroxysafflor yellow A, gentiopicroside, ferulic acid and liquiritin in 15 batches of material benchmarks of Shentong Zhuyutang.Method:The quantitative analysis was carried out on a Thermo Hypersil GOLD C18 column (4.6 mm×250 mm, 5 μm) with mobile phase of acetonitrile-0.1%phosphoric acid aqueous solution for gradient elution, the flow rate was 1.0 mL·min-1, the detection wavelengths were set as 248 nm (0-11 min, inosine), 235 nm (11-14 min, loganic acid), 324 nm (14-16 min, chlorogenic acid), 220 nm (16-19 min, amygdalin and hydroxysafflor yellow A), 274 nm (19-26 min, gentiopicroside), 247 nm (26-54 min, ferulic acid and liquiritin), the column temperature was maintained at 25 ℃. According to the contents of eight active components in 15 batches of material benchmarks, orthogonal partial least squares discriminant analysis (OPLS-DA) in SIMCA 14.1 was used to evaluate the quality difference of each batch of samples.Result:Each component had good separations, the linear ranges of the above 8 components were 2.1-67.2, 1.812 5-58, 1.937 5-62, 5.212 5-166.8, 8.45-270.4, 7.075-226.4, 1.775-56.8, 3.875-124 mg·L-1, respectively (r≥0.999 6). The average recoveries of them were 99.23%, 100.09%, 99.33%, 98.85%, 99.15%, 98.75%, 99.42%, 98.96%, respectively (RSD<2%). The contents of the above eight components in 15 batches of material benchmarks were 0.183 5-0.250 3, 0.173 1-0.265 3, 0.069 5-0.169 8, 0.959 2-1.458 2, 1.905 4-2.553 3, 0.933 3-1.997 5, 0.084 6-0.143 4, 0.212 5-0.704 3 mg·g-1, respectively. Liquiritin, ferulic acid, gentiopicroside and hydroxysafflor yellow A were determined to have significant impact on the quality of different batches of material benchmarks of Shentong Zhuyutang through OPLS-DA.Conclusion:The established method for simultaneous determination of multi-components is reliable, simple and in line with the requirements of methodological verification. It is suitable for the quality control of research and development of compound preparations of Shentong Zhuyutang.
Keywords：Shentong Zhuyutang;classical famous formulas;wavelength switching method;active ingredients;hydroxysafflor yellow A;liquiritin;orthogonal partial least squares discriminant analysis
Abstract：Objective:To explore the therapeutic effect and mechanism of Chaige Qinlian Tang on pneumonia in young mice.Method:The pneumonia model was duplicated by slowly dripping Staphylococcus aureus into the nasal cavity of mice.After successful modeling, the mice were randomly divided into model group, clindamycin group, and high and low-dose Chaige Qinlian Tang groups, with sham operation group as negative control group.The rats were given 200 mg·kg-1 high-dose Chaige Qinlian Tang, 100 mg·kg-1 low-dose Chaige Qinlian Tang and 120 mg·kg-1 clindamycin.The mice were observed every day.Colonies were counted in the lungs of each group five days later.The expression levels of interleukin(IL)-16, tumor necrosis factor (TNF)-α in lung lavage fluid of each group were determined by enzyme linked immunosorbent assay (ELISA). Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to measure the expression levels of IL-16, TNF-α in lung lavage fluid of each group.The expressions of tumor necrosis factor receptor (TNFR) 1, Caspase-3 and Caspase-7 in lung and the pathological changes of lung were observed.Result:Compared with the sham operation group, the respiratory state and the activity state of the model mice were worse, and the survival rate was higher in the high-dose Chaige Qinlian Tang group.Compared with the sham operation group, the pulmonary colony counts in the model group and treatment groups were increased, compared with the model group, the lung colony counts in clindamycin group and high-dose Chaige Qinlian Tang group were improved significantly (P<0.05, P<0.01). Compared with the control group, the expression levels of IL-16, TNF-α, TNFR1, Caspase-3, Caspase-7 mRNA and protein in the lung of model group and treatment groups were significantly increased (P<0.01). Compared with model group, the expression levels of IL-16, TNF-α and TNFR1, Caspase-3, Caspase-7 in the lung of clindamycin group and high and low-dose Chaige Qinlian Tang groups were significantly increased (P<0.01). The expression levels of protein and mRNA were significantly decreased (P<0.05, P<0.01), and the pathological changes of lung were improved, especially in clindamycin group and high-dose Chaige Qinlian Tang group.Conclusion:Chaige Qinlian Tang has a certain therapeutic effect on Staphylococcus aureus pneumonia in young mice.This effect may be related to regulating TNFR1, Caspase-3 and Caspase-7 pathways, reducing the secretion of IL-16 and TNF-alpha, and enhancing the clearance of staphylococcus aureus.
Abstract：Objective:To investigate the protective effect of modified Yinchenhao Tang on α-isothiocyanate(ANIT)-induced cholestatic liver disease (CSLD).Method:Wistar rats were randomly divided into 7 groups: blank control group, model control group, compound Glycyrrhizin capsules group(22.5, 45 mg·kg-1), modified Yinchenhao Tang low, middle and high dose groups(4.1, 8.1, 16.2 g·kg-1). A model of cholestatic liver injury was prepared by intragastric administration of ANIT (100 mg·kg-1). Glycyrrhizin capsules and modified Yinchenhao Tang were administered intragastrically on the second day of modeling for 4 consecutive days. And bile duct intubation was performed on the fifth day to measure the bile flow rate of the rats, and serum was taken to test the total bilirubin(TBIL), direct bilirubin(DBIL), indirect bilirubin(IBIL), alanine aminotransferase(ALT) and total bile acid(TBA) serological indicators of each group. Pathological changes of liver tissues were observed by hematoxylin-eosin (HE) staining. The expression levels of G protein-coupled bile acid receptor(TGR5), nucleotide binding oligomerization domain-like receptor 3(NLRP3) and cysteinyl aspartate specific proteinase-1(Caspase-1) proteins in the iver tissues were detected by Western blot.Result:Compared with the blank control group, bile flow rate in the model group decreased significantly(P<0.01). TBIL, DBIL, IBIL, ALT and TBA level in serum were significantly increased(P<0.01), liver tissue lesions were severe, and significantly increased the expression of liver tissue TGR5 and Caspase-1.Compare with model group, the compound Glycyrrhizin capsules group had no significant effect on bile flow rate and TBIL, DBIL, IBIL, ALT and TBA level in serum. Bile flow rate increased and TBIL, DBIL, IBIL, ALT and TBA level in serum decreased significantly in modified Yinchenhao Tang high dose group. The compound Glycyrrhizin capsules group and modified Yinchenhao Tang group have different extents of improvement the pathological changes of the lung tissues, and the protein expression of TGR5 and Caspase-1 were significantly decreased in the liver tissue(P<0.01).Conclusion:Modified Yinchenhao Tang can effectively treat CSLD in rats, and its mechanism may be related to bile acid and bile acid receptor TGR5-mediated inflammatory factors.
Keywords：cholestatic liver disease;modified Yinchenhao Tang;α-isothiocyanate;G protein-coupled bile acid receptor 1;nucleotide binding oligomerization domain-like receptor 3;cysteinyl aspartate specific proteinase-1
Abstract：Objective:To investigate the effects of modified Buwangsan on the learning and memory ability of Alzheimer's disease (AD) model rats and the expression of NOD-like receptor 3 (NLRP3), cysteine-containing aspartate-specific proteases 1 (Caspase-1) and interleukin-1 beta (IL-1β) in NLRP3 inflammatory pathway in hippocampus of AD model rats, and exploring the underlying mechanism of modified Buwangsan.Method:The 52 eligible rats were randomly divided into sham control group, AD model group, low-dose modified Buwangsan group (1.5 g·kg-1) and high-dose modified Buwangsan group (3 g·kg-1). AD mouse model was established by bilateral hippocampus injection of Aβ1-425 μL (2 g·L-1). The rats in low-dose and high-dose modified Buwangsan group received low and high dose modified Buwangsan respectively within the next 4 weeks, once daily. The learning and memory ability was tested by Morris water maze. The expression of NLRP3, Caspase-1 and IL-1β mRNA was tested by quantitative PCR(Real-time PCR) and Western blot.Result:As compared with the sham group, the learning and memory ability of the rats were significantly impaired (P<0.05). Compared with AD model group, the learning and memory ability and the expression levels of NLRP3, Caspase-1, and IL-1β mRNA and protein were all no statistical differences in low-dose modified Buwangsan group, while the learning and memory ability of the rats were significantly improved and the expression of NLRP3, Caspase-1 and IL-1β mRNA in hippocampus of rats was significantly decreased in high-dose modified Buwangsan group (P<0.05).Conclusion:High-dose modified Buwangsan could attenuate neuroinflammation in the hippocampus of AD mouse model via inhibiting the expression of NLRP3, Caspase-1 and IL-1β, which may be the mechanisms of modified Buwangsan could be used to ameliorate the learning and memory ability of AD mouse model.
Abstract：Lung cancer is currently the leading malignant tumor in China, which seriously endangers people's health. Nowadays, traditional Chinese medicine (TCM) has played an increasingly important role in the comprehensive treatment of lung cancer. It has unique advantages in promoting postoperative recovery, reducing the side effects of radiotherapy and chemotherapy, and effectively prolonging the lifetime of patients. Qingzao Jiufei Tang is mainly used to treat the syndrome of Qi-Yin deficiency due to dryness-heat injury to the lungs. The experimental and clinical studies have confirmed that Qingzao Jiufei Tang has a good anti-lung cancer effect and broad application prospects. In this paper, we reviewed relevant literatures through China National Knowledge Infrastructure (CNKI), Weipu Data, Wanfang Data, PubMed and other databases in recent years, and found a few reports on the anti-lung cancer effect of Qingzao Jiufei Tang. There was still a lack of systematic and comprehensive explanation for its specific mechanism of action against lung cancer. This paper systematically summarized the clinical application of Qingzao Jiufei Tang against lung cancer in recent years, as well as its effects through cell-related signaling pathways and energy metabolism against lung cancer cells. It is clear that this decoction can significantly inhibit the signaling pathways of epithelial growth factor receptor (EGFR), nuclear transcription factor kappa B (NF-κB)/intercellular cell adhesion molecule-1 (ICAM-1), and Janus kinase 2 (JAK2)/signal transducers and activators of transcription 3 (STAT3) of the lung cells. It could also inhibit energy metabolism of tumor cells, and reduce the production of relevant metabolites. This will provide new ideas for the clinical application of Qingzao Jiufei Tang against lung cancer.
Abstract：Objective:To observe the effect of Qingzao Jiufei Tang on apoptosis of lung cancer, Janus protein tyrosine kinase 2/signal transducers and transcriptional activator protein 3 (JAK2/STAT3) signaling pathway, as well as the expressions of downstream apoptosis-related proteins Bcl-2-associated X (Bax) and Cyclin D1.Method:Totally 50 male C57BL/6J mice were randomly divided into five groups: chemotherapy group (CTX), model group, high-dose Qingzao Jiufei Tang group, middle-dose Qingzao Jiufei Tang group and low-dose Qingzao Jiufei Tang group, with 10 mice in each group. The model of lung cancer was established by injecting Lewis lung cancer cells into the right axillary of mice. High-dose, middle-dose and low-dose Qingzao Jiufei Tang groups were orally given drugs (11, 5.5, 2.75 g·kg-1·d-1) two weeks before the modeling. Chemotherapy group was administered intraperitoneally at a dose of 50 mg·kg-1·(2 d)-1, while model group was administered intragastrically with the equal volume of normal saline. After inoculation, the mice in each group were continued to be administered. Two weeks later, the mice in each group were killed, and the tumors were collected. Then the JAK2 protein phosphorylation level was detected by immunohistochemistry (IHC). STAT3, Bax and Cyclin D1 protein expression levels were detected by Western blot, and apoptosis of lung cancer cells was observed by transmission electron microscopy.Result:Compared with model group, the phosphorylation levels of JAK2 and STAT3 protein in lung cancer cells were significantly decreased, the expression of Bax protein was significantly increased, and the expression of Cyclin D1 protein was significantly decreased in high-dose Qingzao Jiufei Tang group, middle-dose Qingzao Jiufei Tang group and chemotherapy group, with statistically significant differences (P<0.05, P<0.01). The results of transmission electron microscopy showed significant apoptotic phenomena in high-dose Qingzao Jiufei Tang group, middle-dose Qingzao Jiufei Tang group, low-dose Qingzao Jiufei Tang group and chemotherapy group compared with the model group.Conclusion:Qingzao Jiufei Tang had an obvious effect in promoting the apoptosis of lung cancer cells. Its mechanism may be related to the inhibition of the phosphorylation of JAK2 and STAT3 protein, the promotion of its downstream Bax protein expression and the inhibition of its downstream Cyclin D1 protein expression.
Keywords：Qingzao Jiufei Tang;lung cancer;Janus protein tyrosine kinase 2 (JAK2)/signal transducers and transcriptional activator protein 3 (STAT3);apoptosis
Abstract：Objective:To study the effect of Qingzao Jiufei Tang on the expression of key limiting enzymes hexokinase 2(HK2), phosphofructokinase 2(PFK2) and pyruvate kinase M2 (PKM2), and the glucose content in Lewis mice colon cancer cells.Method:A total of 50 male C57BL/6J mice were randomly divided into model group, chemotherapy group, and high, middle and low-dose Qingzao Jiufei Tang groups, with 10 mice in each group. The lung cancer cell model was established by injecting Lewis lung cancer cells into the right axilla. The high, middle and low dose groups were administered at the doses of 11, 5.5, 2.75 g·kg-1·d-1 for 2 weeks before modeling. The drug was administered through intraperitoneal injection at a dose of 50 mg·kg-1·(2 d)-1 in the chemotherapy group. The model group was intragastrically administered with an equal volume of normal saline. After the inoculation, the drug was administered for two weeks. Two weeks later, all of the mice were put to death, and tumor tissues were collected. The mRNA expression of HK2 was detected by Real-time PCR. the protein expression of PFK2 was detected by Western blot, the PKM2 activity was detected by enzyme-linked immunosorbent assay (ELISA).Result:Compared with the model group, mRNA expressions and activity of PKM2 in lung cancer cells of treatment groups were significantly declined, and glucose content increased significantly, with significant differences from those of model group (P<0.01). The PFK2 protein expressions in lung cancer cells of treatment groups (high, medium and low-dose groups) were significantly decreased (P<0.05, P<0.01).Conclusion:Qingzao Jiufei Tang could inhibit Lewis proliferation, and decrease the glucose intake in lung cancer cells. The effect targets may be the key rate-limiting enzymes HK2, PFK2, PKM2.
Abstract：Objective:To discuss the effect of Qingzao Jiufei Tang on enzymatic activity and regulatory factor of glucose 6-phosphatedehydrogenase(G6PD) in pentose phosphate energy metabolism pathway in lung cancer.Method:Fifty male C57BL6J mice were randomly divided into five groups. Animal models were induced through axillary injection with Lewis cells. The Qingzao Jiufei Tang group was given drugs (11, 5.5, 2.8 g·kg-1·d-1) two weeks before modeling, the cyclophosphamide(CTX) group was intraperitoneally injected with CTX (50 mg·kg-1), and the model group was intraperitoneally injected with the same volume of saline after molding. At 14 d after modeling, the mice were sacrificed, and the tumor tissues were collected. The enzymatic activity of G6PD, content of reactive oxygen species (ROS) were detected by enzyme-linked immunosorbent assay(ELISA) method. Expressions of gp91phox and p22phox mRNA were detected by real-time fluorescent quantitative polymerase chain reaction(Real-time PCR) method.Result:Compared with the model group, the enzymatic activity of G6PD in high-dose group, medium-dose group and low-dose group were reduced obviously (P<0.05, P<0.01). Content of ROS, mRNA expressions of gp91phox and p22phox in high-dose group, medium-dose group and low-dose group were reduced obviously (P<0.01).Conclusion:Qingzao Jiufei Tang may inhibit the expression of G6PD by inhibiting the expression of gp91 phox, p22phox oxidase, and then reduce content of ROS, so as to reduce the energy metabolism and hyperplasia of lung cancer cells.
Keywords：Qingzao Jiufei Tang;pentosephosphate;regulatory factor;glucose-6-phosphate dehydrogenase (G6PD);reactive oxygen species (ROS)
Abstract：Objective:To clarify the inhibitory effect of essential oil from Alpinia zerumbet rhizome (EOFAZ) on oxidized low-density lipoprotein (ox-LDL)-induced transformation of macrophage into foam cell and explore its possible mechanism.Method:THP-1 monocyte was incubated with 100 μg·L-1 phorbol myristate acetate (PMA) to grow into macrophage, experiment was divided into 4 groups as follows, control group, model group (80 mg·L-1 ox-LDL), EOFAZ at low dose (80 mg·L-1 ox-LDL+ 4 μg·L-1 EOFAZ)and EOFAZ at high dose (80 g·L-1 ox-LDL+ 20 μg·L-1 EOFAZ). Mathye thiazolye telrazliurn (MTT) method was employed to examine the influence of EOFAZ on macrophage viability. Western blot was used to analyze the expression level of cluster of differentiation 36(CD36) and ATP-binding cassette transporter A1(ABCA1) protein in macrophage. Enzyme-linked immunosorbent assay (ELISA) was used to detect cholesteryl ester contents in macrophage. Oil red O staining was applied to determine the accumulation of lipids in macrophage.Result:EOFAZ showed non-toxic effect on macrophage. Compared to control group, macrophage in model group displayed higher level of cholesteryl ester and lipid droplet(P<0.01), as well as significant increasing of CD36 expression (P<0.01), but no effect on ABCA1 expression. EOFAZ notably reduced the contents of lipids and cholesteryl ester(P<0.01), down-regulated expression of CD36 and up-regulated expression of ABCA1 in macrophage in comparison with the model group(P<0.01), indicating that EOFAZ inhibited transformation of macrophage into foam cell.Conclusion:EOFAZ could inhibit ox-LDL-induced transformation of macrophage into foam cell, the underlying mechanism may involves its ability to increase CD36 expression and decrease ABCA1 expression in macrophage.
Keywords：essential oil from Alpinia zerumbet rhizome;macrophage;foam cell;THP-1 monocyte;cluster of differentiation 36(CD36);ATP-binding cassette transporter A1(ABCA1);atherosclerosis
Abstract：Objective:To observe the effect of berberine and 6-shogaol, main components of Coptiae Rhizoma and Zingiberis Rhizoma, on the inflammatory signaling pathway of Toll-like receptors 4 (TLR4)/nuclear factor kappa B (NF-κB) in colonic epithelial cells of mice with ulcerative colitis.Method:Fifty Kunming mice were randomly divided into normal group, model group, berberine group (100 mg·kg-1), 6-shogaol group (100 mg·kg-1), and 6-shogaol combined with berberine group (200 mg·kg-1), with 10 mice in each group. A mouse model of ulcerative colitis was established through oral administration with 2% dextroan sulfate for two weeks. Each group was given corresponding drugs by gavage, while normal group and model group were given equal amount of normal saline. Serum and colon tissue samples were taken 20 days after administration. Enzyme-linked immunosorbent method was used to detect serum interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) expressions, and real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot method were used to detect TLR4, NF-κB p65 mRNA and protein expressions in colon epithelial tissue.Result:Compared with the normal group, relative expressions of TLR4 and NF-κB p65 mRNA and protein were increased in the model group (P<0.01), and the contents of serum IL-1β and TNF-α were increased (P<0.01). Compared with the model group, relative expressions of TLR4 and NF-κB p65 mRNA and protein were significantly decreased in 6-shogaol group, berberine group and 6-shogaol combined with berberine group (P<0.01), and the contents of serum IL-1β and TNF-α were significantly decreased (P<0.01). Among the three groups, 6-shogaol combined with berberine group had the strongest effect (P<0.01).Conclusion:Both 6-shogaol and berberine can inhibit colonic inflammation, reduce inflammatory damage and treat ulcerative colitis. The combined application of 6-shogaol and berberine has a significant synergism effect. The mechanism is related to the excessive activation of TLR4/NF-κB pathway and the regulation of non-controllable intestinal inflammation.
Abstract：Objective:To investigate the effect of Shenqi Fuzheng injection on the sensitivity of cisplatin through immunomodulation, in order to explore its mechanisms.Method:The cell survival was measured by thiazolyl blue tetrazolium bromide (MTT), the rate of apoptosis was detected by flow cytometry, the expressions of apoptosis-related proteins were detected by Western blot, and the expression level of cytokines was detected by enzyme linked immunosorbent assay (ELISA).Result:Shenqi Fuzheng injection 1, 10, 100 mL·L-1 significantly inhibited the growth of co-cultured cells, and the cell survival rate was 71.8%and 59.9%at the concentration of 10, 100 mL·L-1 respectively.Shenqi Fuzheng injection 10 mL·L-1 combined with cisplatin significantly increased the sensitivity of co-cultured cells to cisplatin.The half maximal inhibitory concentration (IC50) of cisplatin was reduced from 30 to 15 μmol·L-1.The rate of apoptosis induced by the combined treatment increased by 15.5%compared with that of cisplatin 15 μmol·L-1 (P<0.05). The expressions of B-cell lymphoma-2 (Bcl-2), B-cell lymphoma-w (Bcl-w) and B-cell lymphoma-extra large (Bcl-xl) were inhibited, and the expressions of Bcl-2-associated X protein (Bax) and protein induced by BH3 domain (Bid) apoptosis were increased (P<0.05). Shenqi Fuzheng injection reduced the release of interleukin-10 (IL-10) and prostaglandin E2 (PGE2) induced by cisplatin (P<0.05).Conclusion:Shenqi Fuzheng injection improves the sensitivity of MDA-MB-231 to cisplatin by regulating immune cells, and plays a synergistic role in inhibiting the proliferation of breast cancer cells.This study provides experimental basis for the prevention and treatment of tumors with Yiqi Fuzheng method, and experimental reference for the study of traditional Chinese medicine in alleviating drug resistance of tumors.
Abstract：Objective:To observe the intervention effect of Yiqi Huoxue recipe (YQHX) on ventricular remodeling in rats with chronic heart failure, in order to explore its mechanism.Method:Among 40 male SD rats, 10 were randomly selected as the sham operation group. The left anterior descending coronary artery ligation was performed to construct the chronic heart failure(CHF) rat model. After modeling, they were randomly divided into model group, captopril group(13.5 mg·kg-1·d-1) and YQHX group (20 g·kg-1·d-1), and orally given the corresponding drugs. After 8 weeks of intervention, cardiac tissues were collected, body mass and heart mass were weighed, and echocardiography were performed to detect the changes in cardiac structure. Masson staining was performed to determine the myocardial interstitial collagen volume fraction. Western blot was used to detect the expression levels of mitochondrial fusion protein optic atrophy 1 (Opa1) and cleavage protein dynamic-related protein 1 (Drpl). The quantitative real-time fluorescence polymerase chain reaction(Real-time PCR)was applied to detect the expressions of Wnt/β-catenin pathway-related factors such as lipoprotein receptor-related protein 6 (LRP6), glycogen synthase kinase-3β (GSK-3β) and β-catenin.Result:Compared with the sham group, the left ventricular wall of the model group was significantly thickened (P<0.05), the cardiac cavity was significantly enlarged, and the content of collagen in the myocardial interstitium was increased (P<0.01). The expression level of Opal decreased, the expression level of Drp1 increased (P<0.05), the mRNA expression level of LRP6, GSK-3, and β-catenin increased (P<0.01). Compared with the model group, YQHX group can reduce ventricular wall thickening, heart chamber enlargement, myocardial interstitial collagen content, up-regulate the low expression of Opa1, but down-regulate the high expressions of Drpl, LRP6, GSK-3β, β-catenin(P<0.05, P<0.01).Conclusion:YQHX can effectively alleviate ventricular remodeling and improve mitochondrial energy metabolism in rats with CHF. The mechanism may be related to the inhibition of Wnt/β-catenin related factors.
Abstract：Objective:To determine whether the main components of Glycyrrhizae Radix et Rhizoma can improve insulin resistance by regulating glycogen synthesis, glycolysis pathway and fatty acid synthesis in myoblasts of L6 rat myoblasts.Method:Insulin resistance (IR) model of L6 rat myoblasts was established through incubation with 0.05 mmol·L-1 palmitic acid (PA) for 9 hours. Normal group, model group, glycyrrhizic acid (GA, 25 μmol·L-1) group, 18β-glycyrrhetinic acid (18β-GA, 25 μmol·L-1) group, isoliquiritigenin (ILG, 25 μmol·L-1) group and isoliquiritin (ILQ, 25 μmol·L-1) group were set up, glucose content in supernatant of cell culture medium was detected by glucose kit, myoblasts glycogen content was determined by glycogen detection kit, protein expression levels of Sterol regulatory element binding protein-1c(SREBP-1c), fatty acid synthetase (FAS) and glycogen synthase kinase3β(GSK3β) were detected by Western blot, and the mRNA expressions of key enzymes in glycolysis were detected by quantitative real-time fluorescence polymerase chain reaction(Real-time PCR).Result:Compared with those in the normal group, the glucose consumption rate was significantly down-regulated in model group (P<0.01), the glycogen content was decreased (P<0.05), the protein expressions of Sterol regulatory element binding protein-1c (SREBP-1c) and fatty acid synthase (FAS) were decreased (P<0.05, P<0.01), the mRNA expressions of fructose phosphate kinase 1 (PFK1), pyruvate kinase (PK) and hexokinase (HK) were down-regulated (P<0.05), and the protein expression of glycogen synthase kinase 3 (GSK3β) protein was increased (P<0.05). Compared with model group, GA, 18β-GA and ILG could significantly increase glycogen content in myoblasts of IR-L6 rats (P<0.05, P<0.01). GA, 18β-GA and ILQ could significantly increase the expression of SREBP-1c (P<0.05, P<0.01), and GA, 18β-GA, ILG and ILQ could significantly increase the expression of FAS (P<0.05, P<0.01), the mRNA expressions of PFK1, PK and HK (P<0.05, P<0.01), and down-regulate the protein expression of GSK3β (P<0.05).Conclusion:The main components of licorice improve the insulin resistance by promoting glycolysis and glycogen synthesis and regulating fatty acid synthesis.
Keywords：main components of Glycyrrhizae Radix et Rhizoma;L6 rat myoblast;insulin resistance;glycogen;glycolysis;fatty acid synthesis
Abstract：Objective:To observe the effect of Fuzheng Kangai (FZKA) decoction combined with gefitinib on the cells proliferation, apoptosis, invasion and metastasis of human lung adenocarcinoma A549 cells in vitro and in vivo, and relevant mechanisms.Method:The A549 cell proliferation of the control group, FZKA decoction groups (0.2, 0.4, 0.8, 1.6, 3.2 g·L-1), Gefitinib groups (10, 20, 40, 60, 80, 100 μmol·L-1) for 24, 48, 72 hours, and FZKA decoction (2 g·L-1) combined with Gefitinib (10 μmol·L-1) groups for 24 hours was detected by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. The changes of cell apoptosis, invasion and metastasis abilities of A549 cells were analyzed by flow cytometry, Wound Healing, transwell invasion assay. Western blot assay was used to examine the protein expressions of cleaved Caspase-3, B-cell lymphoma-2 (Bcl-2), B-cell lymphoma-2 associated X (Bax), F-box and WD repeat domain-containing (FBW7) and myeloid cell leukemia-1 (MCL-1) in vitro.Result:Compared with control group, FZKA decoction group and Gefitinib group inhibited the cell proliferation, cell apoptosis, cell invasion and metastasis abilities in a dose-dependent and time-dependent manner, and improve the protein expressions of Bax, Caspase-3, FBW7, but decreased the protein expressions of Bcl-2, MCL-1 (P<0.05). Compared with treatment with Gefitinib alone, FZKA combined with Gefitinib inhibited the proliferation of A549 cells, and induced apoptosis more significantly (P<0.05). Compared with treatment with Gefitinib alone, the cell scratch healing and invasion abilities were significantly reduced after combined treatment (P<0.05). FZKA decoction combined with Gefitinib up-regulated Bax, Caspase-3 and FBW7 protein expressions, and down-regulated Bcl-2 and MCL-1 protein expressions compared with treatment with Gifitinib alone (P<0.05).Conclusion:FZKA decoction combined with Gefitinib can inhibit the proliferation, invasion and metastasis, and induce apoptosis on A549 cells. The mechanism may be associated with the FBW7/MCL-1 pathway.
Keywords：Fuzheng Kangai decoction;Gefitinib;synergistic effect;F-box and WD repeat domain-containing(FBW7)/myeloid cell leukemia-1(MCL-1)
Abstract：Objective:To explore the clinical efficacy of modified Bushen Huoxuetang on kidney deficiency and blood stasis type early unexplained recurrent abortion and its effect on intestinal flora.Method:Totally 90 patients with kidney deficiency and blood stasis type early unexplained recurrent abortion were selected from March 2017 to October 2018.According to the random number table, they were divided into control group and observation group, with 45 cases in each group. The control group was given Bushen Huoxue capsule, while the observation group was given modified Bushen Huoxuetang. After treatment, the clinical efficacy, traditional Chinese medicine (TCM) syndrome scores and adverse reactions of two groups were compared, and the changes of serum inflammatory factors, coagulation function and intestinal flora were detected before and after treatment.Result:After treatment, the total effective rate of the observation group was 91.11%, which was higher than 77.78%of the control group (P<0.05). The scores of TCM syndromes in observation group were significantly lower than that in control group (P<0.05). The levels of C-reactive protein (CRP), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interleukin-8 (IL-8) in the observation group were significantly lower than those in control group (P<0.05). The thromboplastin time (APTT), thrombin time (TT) and prothrombin time (PT) in observation group were significantly higher than those in control group, while fibrinogen (FIB) was significantly lower than that in control group (P<0.05). The numbers of enterococcus, yeast and Enterobacter in observation group were significantly lower than those of control group, while the numbers of bifidobacteria and Lactobacillus in observation group were significantly higher than those of control group (P<0.05). The incidence of adverse reactions in observation group was 11.11%, which was lower than 28.89%of the control group (P<0.05).Conclusion:Modified Bushen Huoxuetang has a good clinical efficacy in treating kidney deficiency and blood stasis type early unexplained recurrent abortion, and can reduce the TCM syndrome score. Its mechanism may be related to the reduction of inflammation, and improvement of coagulation function and intestinal flora, with a good safety.
Abstract：Objective:To observe the effect of Jianpi Qingchang decoction on hormone withdrawal in patients with hormone-dependent spleen deficiency damp-heat ulcerative colitis.Method:The 60 patients with hormone-dependent ulcerative colitis with spleen deficiency and damp-heat were selected and collected from the outpatient department and the inpatient department of Longhua Hospital, Shanghai University of Traditional Chinese Medicine(TCM) between April 1, 2012 and January 31, 2014.All of patients in two groups were treated with standard hormone reduction method. The control group was given orally Bupi Yichang pills, 6 g/time, 3 times/day, and the experimental group was given orally Jianpi Qingchang recipe, 300 mL water decoction, 1 dose/d, 2 times/d. All of the patients received continuous treatment for 3 months. After treatment, disease activity index, mucosal healing evaluation, curative effect changes of TCM syndromes and changes of inflammatory factors in the two groups were observed.Result:Compared with before treatment, the two groups of Mayo scores after treatment were significantly reduced (P<0.01), and the experimental group was reduced more significantly than the control group (P<0.01). After treatment, 66.67%of patients in experimental group were in remission, and 13.33%of patients in control group were in remission, with statistically significant differences between two groups (P<0.01). After treatment, the healing rate of the control group was 46.67%, while that of the experimental group was 70.0%, with statistically significant differences (P<0.01). After treatment, the effective rate of TCM syndromes in control group was 80.0%, while that in experimental group was 96.67%, with statistically significant differences (P<0.01). Compared with control group before treatment, the levels of IL-1 in both groups were decreased (P<0.05), while the levels of IL-6 and IL-10 were increased (P<0.05). Compared with control group after treatment, the changes in experimental group were significantly better than those in control group (P<0.05).Conclusion:Jianpi Qingchang decoction can reduce the index of disease activity in patients with hormone-dependent ulcerative colitis due to spleen deficiency and dampness-heat, promote mucosal healing and improve the curative effect of TCM symptoms, and thus is worthy of clinical promotion.
Abstract：Objective:To observe the clinical efficacy of Sanju Kechuan oral liquid on phlegm-turbid obstruction of lung syndrome of asthma at acute episode, in order to study its effect on pulmonary function and inflammatory factors.Method:One hundred and seventeen patients were randomly divided into control group (57 cases) and observation group (60 cases) by random number table. Patients in control group got budesonide suspension for inhalation by atomizer, 1-2 mg/time, 2 times/days. Patients who could not control were added with salmeterol xinafoate and fluticasone propionate powder for inhalation in every morning and evening, 50-250 μg/time. In addition to the therapy of control group, patients in observation group was added with Sanju Kechuan oral liquid, 20 mL/time, 3 times/days. The course of treatment was 7 days. And before and after treatment, the severity of asthma at acute episode and the phlegm-turbid obstruction of lung syndrome were scored. And 1-second forced expiratory volume (FEV1), diurnal variability of peak expiratory flow (PEF), fractional exhaled nitric oxide (FeNO), peripheral blood, percentage of sputum eosinophils (Eos) were recorded, and levels of interleukin-2 (IL-2), interferon-gamma (IFN-γ), IL-4, IL-5 and IL-17 were detected.Result:The clinical efficacy in observation group was better than that in control group (Z=1.916, P<0.05). Scores of severity of asthma at acute episode and phlegm-turbid obstruction of lung syndrome were lower than those in control group (P<0.01). And FEV1 was higher than that in control group (P<0.05), while PEF was lower than that in control group (P<0.05). Asthma control in observation group was better than that in control group (Z=2.231, P<0.01). And levels of FeNO, Eos in sputum, Eos in peripheral blood, IL-4, IL-5 and IL-17 were lower than those in control group (P<0.01), whereas levels of IL-2 and IFN-γ were higher than those in control group (P<0.01).Conclusion:In addition to the therapy of routine western medicine, Sanju Kechuan oral liquid can alleviate the severity of phlegm-turbid obstruction of lung syndrome of asthma at acute episode, improve the lung function, control the attack of asthma and inhibit the airway inflammation, with a better the clinical efficacy than pure western medicine treatment.
Abstract：Objective:To observe the clinical efficacy of Changyanqing mixture on chronic recurrent ulcerative colitis (UC) with damp-heat syndrome of large intestine and the effect on the recurrence of disease, in order to discuss the mechanism of action in terms of the neuro-endocrine-immune inflammation network.Method:One hundred and twelve patients were randomly divided into control group (55 cases) and observation group (57 cases) by random number table. Patients in control group got mesalazine slow release tablets, 0.1 g/time, 4 times/days, and those the score of Mayo≥7 were added with prednisone acetate tablets, 0.75 mg·kg-1·d-1, and bifidobacterium viable powder with warm water after dinner, 1 pack/day, 2 times/days. In addition to the therapy of control group, patients in observation group were also given Changyanqing mixture in the morning and evening, 1 pack/day. A course of treatment was 6 weeks, and patients got further consultation once a week. During the remission stage, patients in both groups got mesalazine slow release tablets, 0.5 g/time, 3 times/days, and patients in observation group were added with Changyanqing mixture until the score of damp-heat syndrome of large intestine reduced by more than 90%. The number of patients entering the remission stage of 6 weeks and the time of remission stage were recorded. Before and after treatment, colonoscopy was detected, and Geboes index, Baron, damp-heat syndrome of large intestine and Mayo were scored. And levels of peripheral blood interleukin-6 (IL-6), IL-8, IL-10, IL-17, vasoactive intestinal peptide (VIP), motilin (MTL) and neuropeptide (NPY) were detected, and relapse at the 24-week follow-up was recorded.Result:After the 6-week treatment, the clinical efficacy in observation group was 100%, which was higher than 89.09%in control group (P<0.05). And the healing rate of mucosa was 96.4%, which was higher than 81.82%in control group (P<0.05). And the response rate in two groups was 100%. At the 6th month after the treatment, the clinical remission rate in observation group was 91.23%, which was higher than 76.36%in control group (χ2=4.581, P<0.05). And the average remission time was shorter than that in control group (P<0.01). After treatment, scores of colonic mucosa, Geboes index, colonic mucosa and Mayo were all lower than those in control group (P<0.01). And levels of IL-6, IL-8, IL-17, VIP and MTL were lower than those in control group (P<0.01), while levels of IL-10 and NPY were higher than those in control group (P<0.01). The relapse rate in observation group was 17.54%, which was lower than 38.18%in control group (χ2=5.955, P<0.05). And the mean recurrence time was longer than that in control group (P<0.01).Conclusion:In addition to the routine western medicine therapy, Changyanqing mixture can alleviate the condition of patients by shortening the course of the disease, reducing the recurrence rate, delaying the recurrence time, and regulating the nerve-endocrine-immune inflammation network.
Keywords：ulcerative colitis;chronic recurrence;damp-heat syndrome of large intestine;Changyanqing mixture;induced remission;recurrence;neuro-endocrine-immune inflammation network
Abstract：Objective:Near infrared spectroscopy was used to detect the concentration density (25 ℃), solid-containing content, rhein content and glycyrrhizic acid content of compound Dahuang decoction.Method:The concentrated liquid of compound Dahuang decoction was determined by near infrared optical fiber transmission spectrometry. The contents of rhein and glycyrrhizic acid were determined by HPLC. Fifty-one samples were used for internal cross-validation, and partial least square regression was used to establish correction models between near-infrared spectrum and density, solid-containing content, rhein content and glycyrrhizic acid content, respectively. Ten unknown concentrated liquid samples were collected for external validation and prediction.Result:The external validation complex correlation coefficients between near-infrared spectra and density, solid-containing content, rhein content and glycyrrhizic acid content of the concentrated liquid of compound Dahuang decoction were 0.995 9, 0.999 6, 0.997 0 and 0.992 2, and the root mean square error of prediction (RMSEP) values were 2.50×10-3, 0.17, 7.57 and 67.10, respectively.Conclusion:The near infrared spectroscopy is suitable for the determination of evaluation indexes of the concentrated liquid index of compound Dahuang decoction, and has the characteristics of rapid, simple, stable and reliable.
Abstract：Objective:To establish a scientific and reasonable grade evaluation standard for Polygoni Multiflori Caulis.Method:The quality constant method was used to conduct a grade evaluation study on Polygoni Multiflori Caulis. A total of 16 batches of samples were collected from Chinese herbal slices enterprises and medicinal materials markets. By measuring its appearance traits (average quality and average thickness of the decoction pieces) and the intrinsic quality indicators (content of 2, 3, 5, 4′-tetrahydroxystilbene-2-O-β-D-glucoside), then its quality constant and percentage quality constant were calculated comprehensively, finally, the grades of Polygoni Multiflori Caulis were classified by quality constant method.Result:The quality constant of 16 batches of the decoction pieces was 0.054-0.417, and the percentage quality constant was 12.98-100.00.If these samples were divided into three grades, the quality constant shall be ≥0.334 for the first-grade decoction pieces, the quality constant shall be <0.334 and ≥0.209 for the second-grade decoction pieces, while for the third-grade decoction pieces, the quality constant shall be <0.209.Conclusion:The grade evaluation method based on quality constant can overcome the shortcomings of traditional evaluation method for decoction pieces, and can realize scientific, objective and simple classification of Polygoni Multiflori Caulis. This study enriches the research data of modern grade evaluation of Polygoni Multiflori Caulis, and provides reference for grade evaluation and market circulation of other decoction pieces.
Abstract：Objective:To improve the quality control system in the production of Jinshuibao capsules, and to provide experimental basis for the follow-up research and application of this preparation.Method:High performance liquid chromatography (HPLC) was employed, the analysis was performed on a Ultimate AQ-C18 column (4.6 mm×150 mm, 5 μm). The chromatographic conditions for the determination of adenosine, guanosine and uridine were as following: mobile phase of methanol-0.1%formic acid aqueous solution for gradient elution, the flow rate of 0.4 mL·min-1, column temperature at 30 ℃, sample quantity of 10 μL, detection wavelength at 260 nm. The chromatographic conditions for the determination of ergosterol were as follows: mobile phase of methanol-water (98∶2), the flow rate of 1 mL·min-1, column temperature at 25 ℃, sample quantity of 10 μL, detection wavelength at 283 nm.Result:The main chromatographic peaks of fermented Cordyceps powder samples in different production stages showed little difference. The linear relationships of adenosine, guanosine and uridine were good (R2 >0.999), their recoveries were 106.06%, 101.25%and 105.88%, respectively. The contents of adenosine and ergosterol in 20 batches of samples extracted from 2016 to 2018 were in line with the requirements in the 2015 edition of Chinese Pharmacopoeia, the contents of guanosine and uridine were 0.97-1.36, 0.67-1.38 mg/capsule, respectively.Conclusion:The quality of Jinshuibao capsules in the market is stable. This method can be used to detect the quality of Jinshuibao capsules, and it is simple, stable and reliable.
Abstract：Objective:To study the correlation between endogenous hormone content, related enzyme activity and embryogenesis during the stratification of Acanthopanax senticosus seeds, and to provide theoretical basis for application of endogenous hormones and related physiological indicators in regulating germination of A. senticosus seeds in production and scientific research.Method:Endophytic fungi as well as different concentrations of nitric oxide and hydrogen peroxide solution were used for soaking the seeds of A. senticosus, and then thermophilic stratification was conducted for the seeds. The content of endogenous hormones such as gibberellin (GA3), abscisic acid (ABA), indole acetic acid (IAA), indole butyric acid (IBA) and salicylic acid (SA) in seeds of A. senticosus were tested by high performance liquid chromatography (HPLC), and the activity change of enzymes in vivo such as catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), and malondialdehyde (MDA) were tested. The correlation between the embryo rate and the hormones and their enzyme activities in the seeds of A. senticosus was studied by the grey correlation method.Result:The embryo rate was significantly positively correlated with IAA, GA3, and CAT, with correlation coefficient of 1.086, 0.935 and 1.067, respectively. There was a significant positive correlation between IAA, GA3, CAT, IBA, and SA, but with a significant negative correlation with ABA, MDA, and POD. The correlation degree was as follows: IAA>CAT>GA3>IBA>SA>SOD>ABA>POD>MDA.Conclusion:IAA, GA3 and CAT have significant promoting effects on embryo rate, and there is a significant correlation between hormone content and enzyme activity, which provides a basis for exploring the seed germination mechanism of A. senticosus.
Abstract：Objective:To study the effect of different nitrogen application rates on the growth and carbon metabolism of Acanthopanax senticosus seedlings, and screen out the rational fertilization conditions, in order to provide basis and guidance for scientific fertilization of artificially cultivated A. senticosus.Method:A single-point, single-factor field experiment was conducted to study the seedlings of growing evenly A. senticosus.Five different nitrogen application treatment groups were set up to treat the seedlings, namely N1 group (30 g·m-2), N2 group (60 g·m-2), N3 group (90 g·m-2), N4 group (120 g·m-2), N5 group (150 g·m-2) and CK group (0 g·m-2), respectively.Three months later, plant height, root circumference, stem circumference, root-shoot ratio and SPAD were measured at harvest time.The contents of protein, sucrose, starch, soluble sugar and reducing sugar in fresh leaves were measured.Result:N3 treatment was the best treatment method for the growth and development of A. senticosus seedlings, and the growth of A. senticosus seedlings was the best under this treatment.The protein content of A. senticosus seedlings in N3 treatment was the highest.Starch and sucrose were best accumulated in N5 treatment group and CK treatment group.N5 treatment had the highest soluble sugar content and reducing sugar content.Conclusion:There is a dose-effect relationship between the growth and development of A. senticosus seedlings; that is to say, low and high nitrogen application treatments will cause stress on A. senticosus seedlings.In conclusion, the suitable nitrogen application rate for A. senticosus growth is 90-120 g·m-2.
Keywords：Acanthopanax senticosus;nitrogen application rate;growth and development;carbon metabolism;carbohydrate
Abstract：Objective:To study the effect of different N application rates on the growth and development of Acanthopanax senticosus and the changes of antioxidant enzyme activity, and screen out the suitable amount of nitrogen fertilizer for its growth and development, in order to provide scientific evidence for rational fertilization of Acanthopanax senticosus in artificial cultivation.Method:By single-point, single-factor field experiment, the study samples were one-year-old seedlings of growing evenly A. senticosus.Five nitrogen application treatment groups were set up in the fields.They were N1 (30 g·m-2), N2 (60 g·m-2), N3 (90 g·m-2), N4 (120 g·m-2), N5 (150 g·m-2) and CK (0 g·m-2) in the control group.Three months later, the raw weight of plant, root, leaf and stem were measured at harvest time.After drying to constant weight, plant dry weight, stem dry weight, leaf dry weight and root dry weight were measured.Fresh leaves of plants were collected to measure malondialdehyde(MDA) content and activities of catalase(CAT), superoxide dismutase(SOD), peroxidase(POD), acid phosphatase(ACP) and aseorbateperoxidase(APX) after harvesting seedlings.Result:The biomass of A. senticosus in group N4 (120 g·m-2) was the highest, the protein content in group N3 (90 g·m-2) was the highest, and the activity of all antioxidant enzymes in group N3 (90 g·m-2) was the lowest.Conclusion:There is a dose-effect relationship between seedlings and the nitrogen application rate.That is to say, low nitrogen application rate and high nitrogen application rate will cause stress on Acanthopanax senticosus, and the activity of antioxidant enzymes under low nitrogen application rate is higher than that under high nitrogen application rate.
Abstract：Objective:To study the effect of plant-soil feedback on the antioxidant enzyme system of Acanthopanax senticosus seedlings, in order to elucidate the changes of plant-soil feedback on the antioxidant enzyme system of A. senticosus seedlings, and provide theoretical basis for revealing the reasons of plant-soil feedback.Method:Through the greenhouse pot experiment, plant height, leaf color value (SPAD), antioxidant enzymes [protein, superoxide dismutase(SOD), catalase(CAT), peroxidase(POD), aseorbateperoxidase(APX), malondialdehyde(MDA)] and yield and growth related indexes of soil without A. senticosus (group 1), soil with A. senticosus (group 2) for three consecutive years and soil with A. senticosus (group 3) for many years were measured respectively.Result:There were significant differences in plant height, SPAD, protein, SOD, CAT, POD, APX and MDA between seedlings of A. senticosus planted for three consecutive years (group 1), two successive years (group 2) and three successive years (group 3). The biomass, MDA, CAT, POD and SOD of A. senticosus seedlings in the soil without A. senticosus (group 1) were higher than those in the soil with A. senticosus (group 2 and 3), while the protein and APX were lower than those in the soil with A. senticosus (group 2 and 3).Conclusion:Plant and soil shows negative feedback regulation during the growth of A. senticosus seedlings, which reduced the activity of antioxidant enzymes. Therefore, the soil without planting A. senticosus has more advantages for the growth of A. senticosus seedlings. The results provide a basis for explaining the effect of plant-soil feedback on the growth of A. senticosus, and a theoretical basis and technical support for the technical standards of A. senticosus cultivation in farmland.
Abstract：Objective:To investigate the effects of plant-soil feedback on secondary metabolites in roots, stems and leaves of Acanthopanax senticosus seedlings.Method:One-year-old seedlings of A. senticosus were planted in the soil where no A. senticosus had been planted before (group 1), soil where A. senticosus had been planted for 3 years (group 2), and soil where A. senticosus had been planted for many years in the greenhouse pot experiment, and the secondary metabolites of its roots, stems and leaves were then analyzed.Result:L-Phenylalanine, 3, 4-dihydroxybenzoic acid, syringin, chlorogenic acid, caffeic acid, eleutheroside E, isofraxidin, rutin, hyperoside, and quercetin had significant differences in leaves and roots of A. senticosus seedlings in the soil of group 3, but there was no significant difference in chlorogenic acid and eleutheroside E in stem. Eleutheroside E, isofraxidin, rutin and hyperoside were not detected in the leaves of seedlings planted in group 3.Most of the secondary metabolites in the roots of A. senticosus seedlings showed positive feedback, while in the stems of Acanthopanax senticosus seedlings, caffeine, A. senticosus glycosides, hypericin and quercetin showed negative feedback, and most of the secondary metabolites in the leaves of A. senticosus seedlings showed positive feedback.Conclusion:The plant and soil showed different feedback in different parts of the growth process of A. senticosus seedlings, and the soil where A. senticosus had not been planted was more advantageous to the secondary metabolites of A. senticosus seedlings. The results of the study provide a basis for the study of the effect of plant-soil feedback on the A. senticosus, and provide the theoretical basis and technical support for the artificial cultivation of A. senticosus.
Abstract：Objective:Sixty-nine germplasm samples of Picria felterrae collected from the main producing areas in Guangxi were subject to genetic diversity and genetic relationship analyses using the simple seguence repeat(SSR) molecular marker technology and good germplasm genes associated with the content of picfeltarraenins were screened so as to provide references for germplasm resource evaluation, phylogenetic analysis, and molecular mark assisted breeding of that species.Method:20 pairs of randomly selected primers were amplified based on the transcriptome sequencing technology. The genetic diversity of and genetic relationship between the 69 samples were analyzed using the genetic polymorphic information for each marker locus, and one-variable linear regression and multiple stepwise regression analyses were performed to screen molecular markers associated with the content of picfeltarraenins.Result:The amplification using the 20 pairs of SSR primers produced 76 alleles, 3.8 alleles for each locus on average, higher than effective alleles (1.969 2), and the rare allele rate was 38.2%, suggesting that the alleles distributed unequally. The polymorphism rates of alleles varied between 0-59%, with an average of 38.24%, showing a great difference among loci. The polymorphic information content (PIC) varied between 0-0.621 1, with an average of 0.378 0.Shannon polymorphic information index varied between 0-1.240 1, with an average of 0.759.Nei's gene diversity index (Nei) varied between 0-0.682 3, with an average of 0.440 9.P21 had the highest level accompanied with the lowest P7 for the above three indexes, and significant genetic diversity differences were identified among the loci. For all loci, the mean observed heterozygosity was 0.382 4, lower than the average expected heterozygosity of 0.442 5, suggesting the loss of heterozygosity, the average genetic differentiation coefficient (Fst) was 0.365 9 and the average gene flow Nm was 0.433 2, suggesting a high genetic divergence and a low gene flow. The results of one-variable linear regression and multiple stepwise regression showed that there were 5 loci related to each of the picfeltarraenin IA and IB, and only 1 loci was associated with the content of both.Conclusion:There were significant differences in the genetic diversity of 20 SSR marker sites, and the 69 germplasm samples were greatly genetically differentiated and had low gene flow. From the selected 20 SSR markers 9 marker loci associated with the content of picfeltarraenin IA and IB were selected. The results can be used as a reference for phylogenetic analysis and selective breeding of Picria felterrae.
Abstract：Objective:To clone the complementary deoxyribonucleic acid (cDNA) of auxin/indole acetic acid protein (Aux/IAA) from Glycyrrhiza glabra (GgARPI) and analyze its sequence by bioinformatics.Method:RNA was extracted from fresh root of G. glabra, the cDNA sequence of GgARPI gene was cloned by reverse transcription polymerase chain reaction (RT-PCR), then sequencing and bioinformatic analysis were performed.Result:The GgARPI cDNA sequence with the full length of 686 bp was obtained from G. glabra. The full open reading frame (ORF) was 585 bp, encoding 194 amino acid residues. The bioinformatic analysis showed that the protein coded by GgARPI was a stable hydrophilic protein, with a relative molecular weight of 21.95 kDa and isoelectric point of 6.85.It contained no signal peptides or transmembrane domain. Its secondary structure mainly consisted of random coil. An Aux/IAA superfamily was included in the conversed domain. Homology analysis indicated that it had a close evolutionary relationship with leguminous plants, and a distant evolutionary relationship with monocotyledon, such as Setaria italica.Conclusion:GgARPI cDNA sequence is successfully cloned from G. glabra for the first time, which will lay a foundation for studying the function of GgARPI and explaining the molecular regulatory mechanism of biosynthesis of glycyrrhizic acid in G. glabra.
Abstract：Objective:To identify the active anti-tumor constituents of Benzoinum according to observation of the anti-tumor effect of chemical constituents from Benzoinum in vitro.Method:The 95%ethanol extract of Benzoinum was systematically separated by silica gel column chromatography, medium pressure liquid preparation chromatography and preparation liquid chromatography, and their structures were identified by physicochemical property and spectral data. Anti-tumor activities of the compounds of Benzoinum were screened by in vitro cells including human hepatoma cells in vitro (HepG2), human lung cancer cells (A549), human cervical cancer cells (HeLa), human breast cancer cells (MCF-7) and human prostate cancer cells (PC-3).Result:Fifteen compounds were isolated from Benzoinum and identified as myricadiol(1), 3-keto-oleanonic acid(2), (4E)-1, 5-bis(4-hydroxyphenyl)-1-methoxy-2-(methoxy-methyl)-4-pentene(3a and 3b), (E)-p-coumaryl alcohol γ-Ο-methyl ether(4), sesamin(5), 5-(3″benzoyloxypropyl)-7-methoxy-2-(3′, 4′-methylenedioxy phenyl)-benzofuran(6), dibutyl phthalate(7), methyl 4-hydroxy-3-methoxybenzoate(8), p-hydroxybenzaldehyde(9), p-hydroxyacetophenone(10), acetovanillone(11), 3-oxo-olean-11, 13(18)-dien-28, 19β-olide(12), vanillin(13), benzoic acid(14), and siaresinolic acid(15). Compounds 1 to 11 were isolated from the resin of Styrax tonkinensis for the first time. A part of these compounds had good anti-tumor activities. Among them, compound 2, 12 showed a strongest activity.Conclusion:The chemical constituents of Benzoinum have good prospects for the development and application of anti-tumor drugs.
Keywords：Benzoinum;chemical constituents;isolation and identification;anti-tumor activity;ethanol extract
Abstract：Objective:To explore the pharmacological mechanism of Xiao Xianxiongtang in treating type 2 diabetes mellitus (T2DM) by network pharmacology.Method:The main active ingredients, corresponding targets and target genes of Xiao Xianxiongtang were searched on Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) website. Relevant target genes of T2DM were obtained through Gene Cards. The targets of drug active ingredients were mapped to the targets of T2DM, and the intersection targets were obtained as the predictive targets of Xiao Xianxiongtang on T2DM. Cytoscape 3.7.1 software was used to construct the drug active ingredient-intersection target network model and select the key active ingredients. Interactive protein-protein interaction network (PPI) was constructed by STRING website, and key target genes were selected. Gene function analysis (GO) and enrichment analysis based on the Kyoto encyclopedia of genes and genomes (KEGG) pathway were performed on the intersecting targets using DAVID6.8 online tool.Result:Xiao Xianxiongtang had 30 active ingredients, 156 relevant targets, 14 key active ingredients and 18 key target genes on T2DM. GO analysis showed that the biological functions of Xiao Xianxiongtang in the treatment of potential genes of T2DM mainly involved transcriptional regulation, oxidative stress, protein binding and inflammatory reaction. KEGG pathway enrichment showed that the main pathways of Xiao Xianxiongtang in the treatment of T2DM were hypoxia inducible factor-1 (HIF-1) signaling pathway, tumor necrosis factor (TNF) signaling pathway, Toll-like receptor signaling pathway and thyroid hormone signaling pathway, phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt) signaling pathway, hepatitis B, hepatitis C, tyrosine kinase receptor2(ErbB) signaling pathway, calcium signaling pathway and nuclear factor-kappa B (NF-κB) signaling pathway. Conclusion: Xiao Xianxiongtang is a multi-component, multi-target and multi-pathway process in the treatment of T2DM. It plays an important role in the treatment of T2DM by regulating transcription, oxidative stress, protein binding and inflammatory reaction.Conclusion:The mechanism of Xiao Xianxiongtang in treating T2DM may alleviate insulin resistance, increase insulin sensitivity and reduce blood sugar by inhibiting the secretion of inflammatory factors, participating in anti-inflammatory response, reducing oxidative stress, increasing intracellular calcium concentration, blocking glucagon signaling pathway and activating PI3K/Akt pathway.
Keywords：Xiao Xianxiongtang;type 2 diabetes mellitus;network pharmacology;mechanism of action
Abstract：Objective:To analyze the potential targets and mechanism of Ginseng Radix et Rhizoma-Astragali Radix treatment in lung cancer based on network pharmacology.Method:The Ginseng Radix et Rhizoma, Astragali Radix ingredients and target genes were screened by the traditional Chinese medicine system pharmacology database and analysis platform (TCMSP). Lung cancer-related target genes were obtained from the human gene database (GeneCards). Cytoscape was used for constructing a " drug-ingredient-target-disease" network. Protein-to-protein interaction (PPI) data was downloaded from STRING and then PPI core genes was constructed by CentiScape. Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis of key target genes was performed using R software.Result:A total of 17 Ginseng Radix et Rhizoma and 16 Astragali Radix ingredients were screened. 50 target genes of Astragali Radix and 95 target genes of Ginseng Radix et Rhizoma in the treatment of lung cancer were obtained. A " drug-ingredient-target-disease" network was constructed. 38 PPI core genes were screened using CentiScape. GO function enrichment showed that biological functions of Ginseng Radix et Rhizoma-Astragali Radix were concentrated in nuclear receptor function, transcription-related function, ubiquitination and apoptosis. KEGG pathway enrichment showed that Ginseng Radix et Rhizoma-Astragali Radix treatment in lung cancer were mainly involved in phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt), apoptosis, tumor necrosis factor (TNF) and other pathways.Conclusion:By constructing a " drug-ingredient-target-disease" network, the mechanism of Ginseng Radix et Rhizoma-Astragali Radix treatment in lung cancer was discussed from the perspective of multi-component, multi-target and multi-pathway, which provides reference for further research.
Keywords：network pharmacology;Ginseng Radix et Rhizoma;Astragali Radix;lung cancer;mechanism
Abstract：The prevalence and mortality of cardiovascular diseases are still on the rise in China. Percutaneous coronary intervention(PCI)is the most effective strategy to reduce coronary artery stenosis and occlusion, it can also improve clinical symptoms on patients with coronary heart disease. Clinically, a large number of patients still have heartache symptoms after recanalization of the great vessel. During the perioperative period, PCI is often accompanied with adverse events such as oxidative stress, inflammation, calcium overload, endothelial function damage, and thrombus obstruction. More specifically, during the process of PCI, oxidative stress is more severe than other damages. For example, during the reperfusion period of PCI, the outbreak of reactive oxygen species (ROS) can lead to abnormal mitochondrial energy transfer and cell ion homeostasis, resulting in endothelial cell injury, it can also increase the level of vascular permeability, albumin leakage, the adhesion of leukocyte and platelet, and the level of vascular stenosis. After PCI, traditional Chinese medicine(TCM) brings vasucular restenosis into the category of " palpitation" . The movement of Qi and bloods is tightly related to the five Zang-orans. When the movement of Qi and blood is stagnant, the function of the heart's blood production and circulation is blocked, resulting in Qi of deficiency and blood stasis, thus causing a state of physical weakness. Moreover, at present, Probucol and Ticagrelor and other first-line clinical drugs have certain limitations, and ignore the body' s current status of " essential illusory and real" , unable to coordinate the relationship between the body' s Qi and blood, Yin, Yang and five zang-organs. The purpose of this research is to perform the negative effects of oxidative stress on coronary artery based on the molecular research of modern medicine accompanied with acknowledgement of TCM and to conclude the process of researching prevention and treatment of TCM, and to provide reference for the treatment of coronary artery and coronary artery experimental study.
Keywords：oxidative stress;damage of percutaneous coronary intervention;molecular mechanism;cognition of traditional Chinese medicine(TCM);TCM prevention and treatment
Abstract：Taohong Siwutang is a classical famous formula for promoting blood circulation and removing blood stasis. This paper reviewed the research progress of chemical constituents, pharmacological activities, clinical applications of Taohong Siwutang in recent years. At present, the study on the chemical constituents of different extracts of Taohong Siwutang is systematic. The study of its pharmacological effects mostly includes promoting blood circulation and removing blood stasis, regulating menstruation, promoting fracture healing, and so on. In clinical practice, Taohong Siwutang can be used in the treatment of multi-system and multi-viscera diseases, such as gynecological diseases, internal diseases, orthopedic diseases, dermatological diseases, and the like. Based on this, the quality markers of Taohong Siwutang are predicted and analyzed from the perspectives of quality transmissibility and traceability, ingredient specificity, component validity, component measurability, and formula compatibility environment, which is called five principles of quality marker (Q-marker). According to the analysis, ferulic acid, paeoniflorin, amygdalin, albiflorin, hydroxysafflor yellow A, catalpol and gallic acid can be selected as Q-markers of Taohong Siwutang. Subsequently, these Q-markers can be selected as indicators to conduct whole quality control of Taohong Siwutang and establish a quality traceable system by the quality transmitting of medicinal materials, decoction pieces, intermediates and corresponding objects, so as to provide a reference for the study of the whole process quality control system of Taohong Siwutang.
Keywords：Taohong Siwutang;quality marker (Q-marker);classical famous formulas;quality control;chemical constituents;pharmacological activities;clinical application
Abstract：The epithelial-to-mesenchymal transition (EMT), a process during which cells undergo transition from a polarized epithelial phenotype to a non-polarized mesenchymal phenotype, executed by transcription factors of Twist, Snail and Zeb families. EMT plays an important role in multiple stages of cancer progression such as initiation, tumor growth, and metastasis. Some active ingredients from Chinese materia medica can inhibit EMT by regulating transcription factors and signaling pathways by multiple targets. However, their therapeutic effect was hindered due to various limitation such as solubility, stability, tissue specificity and safety. Therefore, in order to improve the druggability of active ingredients from Chinese materia medica, enhance the therapeutic effect in inhibiting tumor metastasis mediated by EMT and reduce the toxic and side effects, a variety of nano-drug delivery systems have been developed in recent years. Here, we made a review about these drug delivery systems modulating EMT and their research progress in inhibiting tumor metastasis.
Keywords：epithelial-to-mesenchymal transition;active ingredients from Chinese materia medica;nano-drug delivery systems;tumor metastasis;invasion;inorganic nanocarriers;signaling pathways
Abstract：Hepatic fibrosis refers to the pathological process of abnormal proliferation of intrahepatic connective tissue caused by various pathogenic factors, resulting in the excessive accumulation of extracellular matrix (ECM) in the liver and the formation of fibrous scar. Its continuous deterioration will gradually develop into liver cirrhosis, liver failure, liver cancer and other serious liver diseases. Because liver fibrosis and early liver cirrhosis can be reversed, it is very important to control the reversible process of liver fibrosis for the prevention and treatment of liver cirrhosis and liver cancer. In recent years, it has been found that traditional Chinese medicine(TCM) has the characteristics of multi-target, less toxic and side effects and good effect in the treatment of liver fibrosis. In this paper, the mechanism of anti-hepatic fibrosis of TCM and its compound was summarized. TCM can regulate transforming growth factor-β (TGF-β), platelet derived growth factor (PDGF), vascular endothelial growth factor (VEGF), connective tissue growth factor (CTGF) and other growth factors to inhibit the activation of (HSCs) and induce the apoptosis of activated HSCs, promote the expression of adiponectin and inhibit the secretion of leptin, inhibit the inflammatory reaction of liver, resist oxidant stress, inhibit the capillarization of hepatic sinusoidal endothelial cells, so as to effectively prevent the progress of liver fibrosis. Therefore, TCM can inhibit the development of liver fibrosis through multi-mechanism and multi-level, and is one of the important means to treat liver fibrosis.
Keywords：traditional Chinese medicine;liver fibrosis;mechanism of action;research progress