Abstract：ObjectiveTo establish the determination for index components in benchmark samples of Erdongtang， and clarify the content and transfer rate rages of index components in 15 batches of benchmark samples， and to explore the quantity transfer of index components of decoction pieces to benchmark samples.MethodFifteen batches of benchmark samples were prepared， the contents of mangiferin， baicalin and glycyrrhizic acid were determined by high performance liquid chromatography （HPLC）-diode array detector （DAD）， the mobile phase was acetonitrile （A）-0.1% formic acid aqueous solution （B） for gradient elution （0-10 min， 10%-17%A； 10-25 min， 17%-19%A； 25-28 min， 19%-25%A； 28-45 min， 25%-33%A； 45-46 min， 33%-45%A； 46-60 min， 45%-55%A）， detection wavelength was set at 254 nm. Contents of timosaponin BⅡ and the sum of protoneodioscin and protodioscin were determined by HPLC-evaporative light scattering detector （ELSD）， the mobile phase was acetonitrile （A）-water （B） for gradient elution （0-20 min， 24%A； 20-25 min， 24%-27%A； 25-33 min， 27%-28%A； 33-36 min， 28%-90%A； 36-41 min， 90%-24%A）.ResultThe methodological verification of the established method was good， which could be used for determination of five index components in benchmark samples. The content ranges of mangiferin， baicalin， glycyrrhizic acid， timosaponin BⅡ， and the sum of protoneodioscin and protodioscin in 15 batches of benchmark samples of Erdongtang were 0.14%-0.23%， 2.40%-3.37%， 0.07%-0.44%， 0.43%-0.95%， and 0.15%-0.47%， the transfer rate ranges of them were 33.90%-52.15%， 84.46%-105.61%， 22.59%-93.86%， 38.07%-61.43%， and 53.28%-96.11%， respectively.ConclusionThe consistencies of transfer rate of mangiferin， baicalin， timosaponin BⅡ and the sum of protoneodioscin and protodioscin （except glycyrrhizic acid） between decoction pieces and benchmark samples of Erdongtang are good， indicates that the transfer rates of 4 index components are stable during the preparation process of benchmark samples， which can provide data support for research and development of the compound preparation of this formula.
Keywords：classical famous formulas;Erdongtang;benchmark samples;high performance liquid chromatography （HPLC）;quantity transfer law;determination;transfer rate
Abstract：ObjectiveTo explore the mechanism of Didangtang （DDT） against the inflammatory cascade triggered by foam cell pyroptosis in high-glucose environment.MethodOxidized low density lipoprotein （ox-LDL， 100 mg·L-1） was used to induce pyroptosis of foam cells. The control group （5.5 mmol·L-1 glucose）， foam cell group （100 mg·L-1 ox-LDL）， high-glucose group （33.3 mmol·L-1 glucose）， DDT group （10% DDT-containing serum）， and NOD-like receptor family pyrin domain-containing 3 （NLRP3） inhibitor group （MCC950， 10 nmmol·L-1） were designed. The cell membrane damage was observed by lactate dehydrogenase （LDH） release assay. The expression of cysteinyl aspartate-specific proteinase-1 （Caspase-1） was detected by immunofluorescence method， and expression of key proteins NLRP3， Caspase-1， gastermin D （GSDMD）， interleukin-1β （IL-1β）， and interleukin-18 （IL-18） in the pyroptosis pathway was determined by Western blot. The release of IL-18 and IL-1β， monocyte chemoattractant protein-1 （MCP-1）， and tumor necrosis factor α （TNF-α） in cell supernatant was measured by enzyme-linked immunosorbent assay （ELISA）.ResultThe expression of NLRP3， Caspase-1， and GSDMD was up-regulated （P<0.01） and the release of IL-1β， IL-18， MCP-1， IL-1α， and TNF-α was increased （P<0.01） in foam cell group compared with those in the control group. The expression of NLRP3， Caspase-1， and GSDMD was higher （P<0.01） and the release of inflammatory factors was more （P<0.01） in the high-glucose group than in the foam cell group. DDT and MCC950 can inhibit expression of NLRP3， Caspase-1， GSDMD and reduce the release of IL-1β， IL-18， MCP-1， IL-1α， and TNF-α.ConclusionDDT can suppress the pyroptosis of foam cells induced by NLRP3/Caspase-1 pathway in high-glucose environment and thereby alleviate the inflammatory cascade.
Abstract：ObjectiveTo study the underlying mechanism of Liuwei Dihuangwan in inhibiting triple-negative breast cancer through mitogen-activated protein kinase kinase kinase 1 （MAPKKK1） and Krüppel-like factor 4 （KLF4）.MethodFour hundreds SPF female Kunming mice aged 11.5 months were palpated once every 3 days. The model mice of spontaneous tumors were randomly divided into a model group （normal saline）， a paclitaxel group （0.025 g·kg-1·d-1， ip， 21 days）， and high-， medium- and low-dose Liuwei Dihuangwan groups （7.2， 3.6， 1.8 g·kg-1·d-1， ig）. Tumor tissues were separated until the moribund stage. The tumor volume and weight were measured， and the tumor inhibition rate and the survival time of the tumor mice were calculated （after 6 months， tumor-free mice were assigned into the normal group）. SPF SD rats were selected to prepare serum samples containing Liuwei Dihuangwan of different concentrations for cell culture， and MAPKKK1 in MDA-MB-231 cells was silenced. The protein expression of MAPKKK1 and KLF4 was detected by immunofluorescence and Western blot.ResultThe in vivo experimental results showed that compared with the conditions of the normal group， the protein expression of MAPKKK1 and KLF4 in tumor tissues of the model group dropped （P<0.01）. Compared with the model group， all medication groups showed reduced tumor volume and weight （P<0.05， P<0.01）， increased tumor inhibition rate， prolonged survival time of tumor mice （P<0.05）， and increased protein expression of MAPKKK1 （P<0.01）. Additionally， the paclitaxel group and the high-dose Liuwei Dihuangwan group exhibited increased protein expression of KLF4 （P<0.01）. The in vitro experiments showed that compared with the conditions of the normal group， the fluorescence intensities of MAPKKK1 and KLF4 in MDA-MB-231 cells in all medication groups were potentiated， and the protein expression of MAPKKK1 in the paclitaxel group and the high- and medium-dose Liuwei Dihuangwan groups， and the protein expression of KLF4 in the paclitaxel group and high-dose Liuwei Dihuangwan group increased （P<0.01）. After silencing of MAPKKK1， compared with the conditions of the negative plasmid group （unsilenced MAPKKK1）， the fluorescence intensities of MAPKKK1 and KLF4 and the protein expression decreased in the RNAi-27 positive plasmid group （silenced MAPKKK1） （P<0.05， P<0.01）. Compared with the RNAi-27 positive plasmid group， all medication groups had enhanced fluorescence intensities of MAPKKK1 and KLF4 and protein expression to different degrees （P<0.05， P<0.01）.ConclusionLiuwei Dihuangwan can inhibit the growth of triple-negative breast cancer， and the underlying molecular mechanism is related to the up-regulation of MAPKKK1 and activation of KLF4 expression.
Keywords：Liuwei Dihuangwan;triple-negative breast cancer;mitogen-activated protein kinase kinase kinase 1 （MAPKKK1）;Krüppel-like factor 4 （KLF4）;plasmid silencing
Abstract：ObjectiveTo explore the mechanism of Wutou Chishizhi Wan in regulating autophagy and phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/glycogen synthase kinase-3β （GSK-3β） signaling pathway in rats with myocardial ischemia-reperfusion injury （MIRI）.MethodSixty male SD rats were randomly assigned into the normal group （normal saline）， model group （normal saline）， positive control （trimetazidine， 5.4 mg·kg-1） group， and low-， medium-， and high-dose （1.63， 4.9， 14.7 g·kg-1， respectively） Wutou Chishizhi Wan groups， with 10 rats in each group. The rats in other groups except the normal group underwent left anterior descending coronary artery ligation for modeling. Electrocardiogram was employed to detect the ST-segment elevation to evaluate the modeling. Hematoxylin-eosin （HE） staining was performed to reveal the damage of myocardial tissue. The levels of aspartate aminotransferase （AST） and creatine kinase （CK） were determined by colorimetry， and those of cardiac troponin T （cTnT） and myoglobin （MYO） by enzyme-linked immunosorbent assay （ELISA）. Western blot was carried out to determine the protein levels of microtubule-associated proteins 1 light chain 3 （LC3）， autophagy-related gene Beclin-1， PI3K， Akt， GSK-3β， p-GSK-3β， and p-Akt.ResultCompared with the normal group， the modeling elevated the serum levels of AST， CK， cTnT， and MYO （P<0.01）， destroyed the arrangement of myocardial cells abd nucle， twisted and broken myocardial fibers， up-regulated the protein levels of LC3Ⅱ/Ⅰ and Beclin-1 （P<0.01）， and down-regulated the protein levels of PI3K， p-Akt， and p-GSK-3β （P<0.01）. Compared with the model group， trimetazidine and Wutou Chishizhi Wan （all the doses） lowered the levels of AST， CK， cTnT， and MYO in serum （P<0.01）， restored the arrangement of myocardial cells and muscle fibers， reduced necrosis， down-regulated the protein level of Beclin-1 （P<0.01）， and up-regulated the protein levels of PI3K， p-Akt， and p-GSK-3β （P<0.01）. Additionally， Wutou Chishizhi Wan （all the doses） down-regulated the protein level of LC3Ⅱ/Ⅰ （P<0.05， P<0.01）. Compared with those in the trimetazidine group， the serum AST level rose in the low-dose Wutou Chishizhi Wan group （P<0.05） and declined in the high-dose group （P<0.01）， and the protein level of Beclin-1 was down-regulated in the medium-dose group （P<0.01）. Additionally， the trimetazidine group had higher protein level of LC3Ⅱ/Ⅰ than medium- and high-dose Wutou Chishizhi Wan groups （P<0.05， P<0.01）， higher protein level of PI3K than low-， medium-， and high-dose groups （P<0.01）， lower protein level of p-Akt than low- and medium-dose groups （P<0.01）， and higher p-GSK-3β protein level than the medium-dose group （P<0.01）.ConclusionDifferent doses of Wutou Chishizhi Wan can ameliorate MIRI， and the high dose has the best effect. Wutou Chishizhi Wan can reduce the activity of myocardial injury markers AST， CK， cTnT， and MYO， and alleviate the pathological damage of myocardial tissue. It can down-regulate the protein levels of beclin-1， LC3Ⅱ/Ⅰ， and up-regulate those of PI3K， p-Akt， and p-GSK-3β. In summary， Wutou Chishizhi Wan may inhibit excessive autophagy and regulate the PI3K/Akt/GSK-3β signaling pathway to exert protective effect on MIRI rats.
Keywords：Wutou Chishizhi Wan;myocardial ischemia-reperfusion;autophagy;phosphatidylinositol 3-kinase （PI3K）;protein kinase B （Akt）
Abstract：ObjectiveTo explore whether Hei Xiaoyaosan can inhibit the inflammatory response in the hippocampi of Alzheimer's disease （AD） rats by regulating and activating the Wnt/β-catenin signaling pathway to improve the cognitive and memory dysfunction.MethodAmong the 90 male Wistar rats， 12 were randomly selected as the blank group （normal saline） and 12 as the sham operation group （normal saline）. For the remainder， amyloid β-protein42 （Aβ42） was injected in the left and right hippocampus to induce AD， and then the AD rats were randomized into model group， low-， medium-， and high-dose Hei Xiaoyaosan groups （corresponding doses of Hei Xiaoyaosan， ig）， and donepezil group （donepezil hydrochloride，ig）， with 12 in each group. The administration lasted 42 days. The pathological changes of hippocampal CA1 region was observed based on Nissl staining. The escape latency on the 1st to 5th day in Morris water maze was recorded and the spatial memory on the 6th day was tested. Enzyme-linked immunosorbent assay （ELISA） was employed to detect the expression of interleukin （IL）-6， IL-10， and tumor necrosis factor-α （TNF-α） in rat hippocampus and serum， Western blotting to examine the protein expression of glycogen synthase kinase-3β （GSK-3β）， β-catenin， and peroxisome proliferator-activated receptor gamma （PPARγ）， and real-time polymerase chain reaction （Real-time PCR） to determine the mRNA expression of rat GSK-3β， β-catenin， and PPARγ.ResultCompared with the blank group， the number of neurons in the hippocampal CA1 area of model group was significantly reduced， the arrangement was uneven， the cell body was damaged more obviously， and the Neisser body was unclear. The treatment group was significantly prolonged （P<0.01）， and the number of crossing stations was significantly reduced （P<0.01）， the levels of IL-10 in serum and hippocampus of rats in the model group were significantly decreased， while the levels of IL-6 and TNF-α were significantly increased （P<0.01）， the GSK-3β protein and mRNA in the model group were significantly increased， and the protein expressions of β-catenin and PPARγ were significantly decreased （P<0.01）， and the difference was more obvious. The number of neurons in the donepezil group was more distributed， neatly arranged， the structure was intact， and the Nissl bodies were clear and definite， the escape latency on the 3rd to 5th days in middle and high dose groups of Hei Xiaoyaosan and the donepezil group was significantly shortened （P<0.01）， the number of crossing platforms increased significantly （P<0.01）， the expression levels of IL-10 in the rat hippocampus and serum were significantly increased， while IL-6 and TNF-α were significantly decreased （P<0.01）， GSK-3β in the rat hippocampus was significantly increased. The expressions of GSK-3β protein and mRNA were significantly decreased， while the expression levels of β-catenin and PPARγ protein and mRNA were significantly increased （P<0.01）. There was no significant difference in each index between the donepezil hydrochloride group and the high-dose Hei Xiaoyaosan group.ConclusionHei Xiaoyaosan can inhibit the inflammatory response in the hippocampus of AD rats by regulating the Wnt/β-catenin signaling pathway， thereby alleviating the cognitive and memory impairment of AD rats.
Abstract：ObjectiveTo explore the therapeutic effect and possible mechanism of Banxia Xiexintang and its disassembled prescriptions in regulating the flora disorder induced by mixed antibiotics in young rats.MethodSeventy male BALB/C young rats were randomly assigned into 7 groups： blank group， model group， Bifidobacterium tetralogy viable tablets （0.68 g·kg-1） group， Banxia Xiexintang （9.1 g·kg-1） group， Xinkai （3.19 g·kg-1） group， Kujiang （1.82 g·kg-1） group， and Ganbu （4.1 g·kg-1） group， with 10 rats in each group. Except the blank group， the other groups were given mixed antibiotics by gavage to induce intestinal flora disorder. After 14 days， the rats in different drug groups were administrated with corresponding drugs by gavage， and those in the blank group and model group with the same amount of normal saline once a day for 14 days. After that， fecal samples were collected aseptically for 16S rDNA sequencing of intestinal flora， and lipopolysaccharide （LPS， 10 mg·kg-1） was injected intraperitoneally to induce inflammatory reaction. The tissue morphology of colonic mucosa was observed via hematoxylin-eosin （HE） staining， and the macrophage infiltration of colonic mucosa was observed via toluidine blue staining and immunohistochemistry. The expression of interleukin-1β （IL-1β）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， tumor necrosis factor-α （TNF-α）， and interleukin-10 （IL-10） mRNA were determined by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）.ResultCompared with the blank group， the modeling changed the intestinal flora structure of the young rats （P<0.01）， damaged the colonic mucosa， reduced the macrophage infiltration， and down-regulated the mRNA levels of IL-1β， IL-6， IL-8， TNF-α， and IL-10 （P<0.01）. Compared with the model group， bifidobacterium quadruple viable tablets， Banxia Xiexintang and its disassembled prescriptions increased the diversity of intestinal flora and the relative abundance of beneficial bacteria such as Bacteroidetes and Firmicutes （P<0.01）. At the same time， they ameliorated colonic mucosal injury （P<0.05， P<0.01）， increased macrophage infiltration （P<0.05， P<0.01）， and up-regulated the mRNA levels of IL-6， IL-8， and TNF-α （P<0.01）. The mRNA level of IL-1β was up-regulated in Bifidobacterium tetralogy viable tablets， Banxia Xiexintang， Kujiang， and Ganbu groups （P<0.01）， and that of IL-10 was up-regulated in Bifidobacterium tetralogy viable tablets， Banxia Xiexintang， Xinkai， and Ganbu groups （P<0.01）.ConclusionBanxia Xiexintang and the disassembled prescriptions can adjust the intestinal flora of young rats exposed to antibiotics and protect the immune barrier of colonic mucosa after intestinal flora disorder. In particularly， the whole prescription of Banxia Xiexintang demonstrates the best performance.
Keywords：Banxia Xiexintang;intestinal flora;antibiotics;intestinal immunity;study of compatibility
Abstract：ObjectiveThis study aims to explore the potential molecular mechanism of Gegen Qinliantang （GQL） in the intervention of atherosclerosis （AS） based on network pharmacology and molecular docking.MethodThe active components and targets of each medicinal in GQL were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， and AS-related genes from 7 databases. Thereby， the anti-AS targets of GQL were screened out. Cytoscape 3.8.0 was employed to construct the "component-target" network， and STRING the protein-protein interaction （PPI） network. Core targets were screened out with CytoNCA. R clusterProfiler was used for Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment of target genes， which were then visualized. Finally， molecular docking of the top ten active components with the core targets of AS was performed and the binding affinity was compared with that between atorvastatin and the core targets.ResultIn the end， 150 active components of GQL， 20 289 AS targets， and 213 common targets were retrieved， and 48 core common targets were screened out. They were mainly involved in the GO terms of nuclear receptor activity， ligand activation， and transcription factor activity and the pathways of fluid shear force and AS， advanced glycation end products-receptor for advanced glycation end products （AGE/RAGE）， interleukin-17 （IL-17）， tumor necrosis factor （TNF）， Toll-like receptor pathways and other signaling pathways closely related to AS. The molecular docking results showed that the effective components of GQL had high binding affinity to core targets of AS， and the binding affinity was even higher than that between the atorvastatin and core targets. The five groups with high binding affinity were puerarin-TNF， baicalein-inducible nitric oxide synthase 2 （NOS2）， puerarin-NOS2， and formononetin-NOS2， wogonin-NOS2.ConclusionThe above result provides new ideas for further exploration of this classical decoction.
Abstract：ObjectiveTo explore the mechanism underlying the intervention of Gegen Qinliantang （GQL） in vulnerable plaques in atherosclerosis （AS） of ApoE-/- mice by regulating the polarization of macrophages.MethodTwelve normal C57BL/6CNC mice were used as the control group， and 60 ApoE-/- mice of the same line were randomized into 5 groups： model group， low-dose， middle-dose， and high-dose GQL groups （GQL-D， GQL-Z， and GQL-G groups， respectively）， and atorvastatin group （western medicine group）. High-fat diet was used for modeling. The control group and the model group were given （ig） equal volume of sterile distilled water， and GQL-D， GQL-Z， GQL-G， and western medicine groups received （ig） corresponding concentration of drugs for 8 weeks. The levels of total cholesterol （TC）， triglyceride （TG）， high-density lipoprotein cholesterol （HDL-C）， and low-density lipoprotein cholesterol （LDL-C） were detected with biochemical methods. The distribution of plaques in the aortic region was observed based on oil red O staining and hematoxylin-eosin （HE） staining. Serum levels of M1 pro-inflammatory factors tumor necrosis factor （TNF）-α and interleukin （IL）-6 and M2 anti-inflammatory factors IL-13 and transforming growth factor （TGF）-β were detected by enzyme-linked immunosorbent assay （ELISA）. Protein expression of macrophage mannose receptor CD206/arginase-1 （Arg-1） and CD206/inducible nitric oxide synthase （iNOS） was determined by double-labeling immunofluorescence， and mRNA expression of aortic Arg-1 and iNOS by real-time polymerase chain reaction （PCR）.ResultLevels of TG， TC， and LDL-C were significantly lower and HDL-C level was significantly higher in the GQL-Z， GQL-G， and western medicine groups than in the model group. As the concentration of GQL rose， the area with plaques gradually shrunk and the color became lighter. The staining areas of the GQL-G group and the western medicine group were the most scattered. The administration groups showed significant increase in the protein levels of Arg-1 and CD206， significant decrease in the protein level of iNOS， significant rise of Arg-1 mRNA level， and significant drop of iNOS mRNA level （P<0.05）.ConclusionGQL intervenes in the vulnerable plaques in AS by improving lipid metabolism， inhibiting macrophage M1 polarization， promoting macrophage M2 polarization， and further improving the inflammatory microenvironment.
Abstract：ObjectiveTo explore the effect of Gegen Qinliantang （GQL） on vulnerable plaque of atherosclerosis based on the macrophage pyroptosis mediated by nuclear factor （NF）-κB/NOD-like receptor protein 3 （NLRP3）/cysteine-aspartic acid protease （Caspase）-1 pathway.MethodA total of 12 normal C57BL/6CNC mice were used as the control group， and 60 ApoE-/- mice of the same line were randomized into 5 groups： model group， low-dose， medium-dose， and high-dose GQL groups （GQL-D， GQL-Z， GQL-G groups， respectively）， and western medicine group. The control group and model group were given （ig） equal volume sterile distilled， and GQL-D， GQL-Z， GQL-G and western medicine groups received （ig） corresponding concentration of drugs for 8 weeks. Aortic plaques were observed based on hematoxylin and eosin （HE） staining. Serum levels of interleukin （IL）-1β and IL-18 were detected by enzyme-linked immunosorbent assay （ELISA）， protein levels of macrophage mannose receptor （CD206）/apoptosis-associated speck-like protein containing a CARD （ASC） and CD206/NLRP3 by double-labeling immunofluorescence， and C-terminal gasdermin D （GSDMD）， N-terminal GSDMD， NLRP3， pro-cysteinyl aspartate specific proteinase 1 （pro-Caspase-1） and NF-κB p65 by Western blot.ResultCompared with the control group， model group demonstrated serious pathological changes， rise of the levels of serum IL-1β and IL-18 and tissue ASC， NLRP3， C-terminal GSDMD， N-terminal GSDMD， pro-Caspase-1， and NF-κB p65， and decrease of CD206 level （P<0.05）. As compared with model group， the administration groups showed alleviation of the lesions in aortic wall， decrease in levels of serum IL-1β and IL-18 and tissue ASC， NLRP3， C-terminal GSDMD， N-terminal GSDMD， pro-Caspase-1， and NF-κB p65， and rise of CD206 level， with significant difference between some groups （P<0.05）.ConclusionGegen Qinliantang alleviates vulnerable plaque of atherosclerosis by regulating NF-κB/NLRP3/Caspase-1 pathway and further relieving macrophage pyroptosis.
Abstract：ObjectiveThe tolerance of C57BL/6 mice to artemisinin-sensitive and -resistant strains of Plasmodium berghei （Pb） K173 and the differences in blood parameters， spleen coefficient and spleen structure during infection were compared to explore whether the artemisinin resistance of Pb would aggravate malaria infection.MethodPbK173 artemisinin-sensitive and -resistant strains were tested in parallel. C57BL/6 mice were randomly divided into 1 control group， 4 artemisinin-sensitive strain groups and 4 artemisinin-resistant strain groups by body weight. Each infection group was simultaneously inoculated （ip） with 1×107 infected red blood cells （iRBCs） of sensitive/resistant strain. For the mice in the survival test group， the body weight was recorded every day post infection， and the tail vein blood smear was collected to calculate the Pb infection rate. In the other infection groups， peripheral blood and spleen were collected on 2， 5 and 9 d after infection. Peripheral blood parameters， spleen coefficient， pathological section of spleen and spleen cells were detected in each group.ResultOn 1-3 d after infection， the infection rate of the resistant strain （0.4±0.0， 0.8±0.1， 1.9±0.4）% was always higher than that of the sensitive strain （0.2±0.1， 0.4±0.1， 1.1±0.3）% （P<0.01）. From the 4th d of infection， the infection rate of the two groups gradually approached. The survival period of the sensitive strain group （20.5±1.2） d was shorter than that of the resistant strain group （23.3±1.4） d （P<0.01）. On the 9th d， the white blood cell count of the sensitive strain group （16.2±1.1）×109 cells/L was higher than that of the resistant strain group （10.6±1.8）×109 cells/L （P<0.01）. Flow cytometry analysis of spleen cells showed that the sensitive strain group （3.6±0.4） demonstrated a higher CD4+/CD8+ value than the resistant strain group （2.3±0.2） on the 9th d （P<0.01）. The spleen of C57BL/6 infected mice was gradually enlarged during infection， and on the 9th d， the resistant strain group （3.1±0.1）% showed a higher spleen coefficient than the sensitive strain group （2.7±0.2）% （P<0.01）. In the early stage of C57BL/6 infected mice， the red pulp of spleen was hyperemic and swollen. On the 9th d， the marginal area of the spleen disappeared and the structure of the red and white pulp was destroyed.ConclusionWithout drug treatment， the protective immune responses of peripheral blood and spleen of C57BL/6 mice were more sensitive to PbK173 artemisinin-sensitive strain. The artemisinin-resistant strain of PbK173 bred with mouse-to-mouse blood transmission and increased artemisinin dose exhibited shortened growth period and reduced toxicity.
Abstract：ObjectiveTo explore the mechanism of Naozhenning on learning and memory ability and neuron damage in hippocampal CA1 region of post-concussion syndrome model rats based on mitochondrial function.MethodMultiple cerebral concussion （MCC） was induced in SPF Wistar rats with the free-fall impact method. Then the model rats were randomly classified into model group （equivalent volume of distilled water）， piracetam （0.43 g·kg-1， ig） group， and low-， medium-， and high-dose NZN （5.4， 10.8， 21.6 g·kg-1， respectively， ig） groups， with 10 rats in each group， and another 10 normal rats were included in the normal control group （equivalent volume of distilled water）. The administration lasted 14 days and then relevant indexes were detected. Morris water maze test was used to observe the changes of learning and memory ability in each group， such as escape latency， residence time in primary quadrant， and times of crossing platform. The pathological changes of hippocampal CA1 region were observed based on hematoxylin-eosin （HE） staining and Nissl staining. The ultrastructure of mitochondria was observed under the transmission electron microscope （TME） and the activity of mitochondrial respiratory chain complex Ⅰ was detected by colorimetry. The content of adenosine triphosphate （ATP） was determined by fluorescence probe and mitochondrial membrane potential （MMP） by fluorescein enzyme-linked fluorescence immunoassay.ResultCompared with the normal control group， the model group showed long escape latency， short residence time in target quadrant， few times of crossing the platform， significant decrease in counts of neurons and Nissl bodies in hippocampal CA1 region， damage of neuronal morphology and mitochondrial structure， and significant reduction of MMP and the content of mitochondrial ATP and respiratory chain complex I （P<0.05， P<0.01）. The NZN groups demonstrated short escape latency， long residence time in target quadrant， increased times of crossing the platform， small number of neurons and Nissl bodies in hippocampal CA1 region， alleviated damage of neuronal morphology and mitochondrial structure， and increase in MMP and the content of mitochondrial ATP and respiratory chain complex I （P<0.05， P<0.01）.ConclusionNZN can improve the learning and memory ability of MCC rats by improving mitochondrial structure and function and alleviating hippocampal neuron injury.
Keywords：Naozhenning;post-concussion syndrome;hippocampus;mitochondrial function;learning and memory
Abstract：ObjectiveWe aimed to investigate the efficacy and mechanism of Yishen Shengjing Prescription （YSP） in the treatment of oligoasthenospermia in rats.MethodThe oligoasthenospermia rat model was established by injection with cyclophosphamide （35 mg·kg-1·d-1） for 5 consecutive days. Rats were randomly assigned into control group （without treating with cyclophosphamide）， model group， low- （YSP-L）， medium- （YSP-M）， and high- （YSP-H） dose （2.91， 5.83， and 11.66 g·kg-1， respectively） groups， Wuzi Yanzongwan （WYW， 1.03 g·kg-1） group， and L-carnitine （0.17 g·kg-1） group， with 8 rats in each group. After 28 days of drug intervention， the body weight， testicular weight， and testicular index of rats were recorded. The sperm quality in epididymis was detected by computer-assisted sperm analysis （CASA） system. Hematoxylin-eosin （HE） staining was employed for observation of testicular tissue morphology. The levels of malondialdehyde （MDA） and superoxide dismutase （SOD） in testicular tissue were detected by colorimetry. The levels of follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and testosterone （T） in serum were determined by enzyme-linked immunosorbent assay（ELISA）. TdT-mediated dUTP nick-end labeling （TUNEL） was employed to detect the apoptosis of testicular cells. The protein levels of B cell lymphoma-2（Bcl-2）， Bax， and cleaved Caspase-3 in testicular tissue were detected by Western blot.ResultCompared with the control group， the model group showed decreased body weight， testicular weight and index， sperm concentration and motility （P<0.01） and increased testicular pathological score （P<0.01）. Compared with the model group， the YSP-M， YSP-H， WYW， and L-carnitine groups showed increased body weight， testicular weight， testicular index， sperm concentration and motility and decreased testicular pathological score. After modeling， the SOD level decreased （P<0.01） while the MDA content increased （P<0.01） in the testicular tissue. YSP-H， WYW， and L-carnitine reversed the SOD and MDA level changes caused by modeling. Compared with the control group， the model group exhibited declined T level （P<0.01） and increased FSH and LH levels （P<0.01）. Compared with the model group， YSP， WYW， and L-carnitine increased the T level （P<0.01） and decreased the LH level （P<0.05， P<0.01）. The apoptosis rate of spermatogenic cells in the model group was higher than that in the control group （P<0.01）， whereas YSP-M， WYW， and L-carnitine reversed such changes （P<0.01）. The model group rats showed decreased expression of Bcl-2（P<0.05） and increased expression of Bax and cleaved Caspase-3 （P<0.05， P<0.01）. Compared the model group， YSP-M， YSP-H， WYW， and L-carnitine up-regulated the Bcl-2 expression and down-regulated the cleaved Caspase-3 expression （P<0.05， P<0.01）.ConclusionYSP improved the sperm quality of oligoasthenospermia model rats by regulating the antioxidant system and sex hormone levels and inhibiting the apoptosis of spermatogenic cells.
Abstract：ObjectiveTo observe the effect of polysaccharides from root， stem， leaf and fruit of Schisandra chinensis on exercise endurance in the aging mice induced by D-galactose.MethodMale ICR mice were randomly assigned into six groups： blank control group， model group， root polysaccharide group， stem polysaccharide group， leaf polysaccharide group and fruit polysaccharide group. The mice were administrated with distilled water or root， stem， leaf and fruit polysaccharide （total sugar content of 35 mg·kg-1） by gavage. Thirty minutes after the administration， the blank control group was subcutaneously injected with normal saline， and the other groups with D-galactose （300 mg·kg-1）， once daily for 6 weeks. The anti-fatigue effects were evaluated by rotarod test， forelimb grip strength test， and weight-loaded swimming test. The fatigue and oxidation indicators such as blood urea nitrogen （BUN）， serum lactic acid （LD）， lactic dehydrogenase （LDH）， creatine kinase （CK）， superoxide dismutase （SOD）， malondialdehyde （MDA）， glutathione peroxidase （GSH-Px）， and reactive oxygen species （ROS） were measured by chemical colorimetry. The protein levels of pro-apoptotic protein B cell lymphoma-2 （Bcl-2）-associated X protein （Bax）， anti-apoptotic Bcl-2 and cleaved cysteinyl aspartate-specific protease-3 （cleaved Caspase-3） in mouse skeletal muscle were detected by Western blot.ResultIn the rotarod test， the time on rod was shorter in the model group than in the blank control group （P<0.01） and the root， stem and fruit polysaccharide groups （P<0.01）. In the forelimb grip strength test， the forelimb grip strength in the model group was lower than that in the blank control group （P<0.01） and the root， stem， leaf and fruit polysaccharide groups （P<0.01）. In the weight-loaded swimming test， the weight-loaded swimming time in the model group was shorter than that in the blank control group （P<0.01） and the root， stem， leaf and fruit polysaccharide groups （P<0.01）. Compared with those in the blank control group， the BUN， LD， LDH and CK levels significantly increased in the model group （P<0.05， P<0.01）. The increases in BUN and LDH levels were decreased by the root， stem and fruit polysaccharides （P<0.05， P<0.01） and those in LD and CK by the root， stem， leaf and fruit polysaccharides （P<0.05， P<0.01）. Compared with the blank control group， the model group showed decreased SOD and GSH-Px activities （P<0.01） and increased MDA and ROS content （P<0.01）. Compared with the model group， the root， stem， and fruit polysaccharide increased the SOD activity （P<0.05， P<0.01） and decreased ROS content （P<0.01）. The root and stem polysaccharides decreased the MDA content （P<0.01） and increased the GSH-Px activity （P<0.05， P<0.01）. Compared with the blank control group， the model group showed up-regulated protein levels of Bax and cleaved Caspase-3 and down-regulated protein level of Bcl-2 （P<0.01）. Compared with the model group， the root， and stem polysaccharides down-regulated the protein levels of cleaved Caspase-3 （P<0.05） and up-regulated protein level of Bcl-2 （P<0.01）.ConclusionThe polysaccharides from the root， stem， leaf， and fruit of S. chinensis have anti-fatigue effect in D-galactose-induced aging mice. The polysaccharides may exert such effect by improving the antioxidant capacity and inhibiting the apoptosis of skeletal muscle cells.
Keywords：polysaccharides of Schisandra chinensis;exercise endurance;D-galactose;aging;anti-fatigue
Abstract：ObjectiveTo study the effect of isoflavones from Sojae Semen Praeparatum （ISSP） on lipid metabolism in atherosclerotic mice， and decipher the underlying mechanism via the peroxisome proliferator-activated receptor gamma/liver X receptor alpha/ATP-binding cassette transporter A1 （PPARγ/LXRα/ABCA1） signaling pathway.MethodFifty ApoE-/- mice were randomly assigned into the model group， western medicine （atorvastatin calcium， 3.03 mg·kg-1） group， and low-， medium-， and high-dose ISSP （2.5， 5， 10 mg·kg-1， respectively） groups， with 10 rats in each group. Atherosclerosis model mice were established by bilateral ovariectomy and feeding high-fat diet. Another 10 ApoE-/- mice receiving ovariectomy and high-fat diet were taken as the sham group. Some mice died of postoperative infection， and finally 6 mice were included in each group. One week after operation， each group was administrated with corresponding drugs or equivalent amount of normal saline. After 12 weeks， the levels of triglyceride （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， and non-esterified fatty acids （NEFAs） in serum and liver tissue were measured. The levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in serum were detected by enzyme-linked immunosorbent assay （ELISA）. Hematoxylin-eosin （HE） staining and oil red O staining were used for observation of aortic plaque formation and liver lipid deposition. The mRNA and protein levels of PPARγ， LXRα， ABCA1， and ATP-binding cassette transporter G1 （ABCG1） in liver were determined by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot.ResultCompared with the sham group， the modeling of atherosclerosis increased the aortic plaque area （P<0.01）， elevated the serum TC， TG， LDL-C， TNF-α， and IL-6 levels （P<0.01）， decreased the level of HDL-C （P<0.01）， increased the liver index （P<0.05） and the levels of TC， TG， and NEFAs in liver （P<0.01）， and caused obvious hepatic fat vacuoles and lipid deposition. In addition， the modeling down-regulated the mRNA levels of PPARγ， LXRα， ABCA1 in liver （P<0.05， P<0.01），and regulated the mRNA and protein levels of ABCG1（P<0.05， P<0.01）. Compared with the model group， atorvastatin calcium and middle-， high-dose ISSP reduced the serum TC， TG， LDL-C， TNF-α， and IL-6 levels （P<0.01）， decreased the liver index （P<0.01）， alleviated the liver fat vacuoles and lipid deposition， and increased the levels of TC， TG， and NEFAs in the liver （P<0.05， P<0.01）. Furthermore， they up-regulated the mRNA and protein levels of PPARγ， LXRα， ABCA1， and ABCG1 in the liver （P<0.05， P<0.01）.ConclusionISSP may regulate lipid metabolism through PPARγ/LXRα/ABCA1 signaling pathway to down-regulate the expression of inflammatory cytokines in serum and alleviate liver lipid deposition， thereby suppressing the formation of atherosclerotic plaque.
Keywords：atherosclerosis;lipid metabolism;isoflavones from Sojae Semen Praeparatum;peroxisome proliferator-activated receptor gamma/liver X receptor alpha/ATP-binding cassette transporter A1 （PPARγ/LXRα/ABCA1） signaling pathway
Abstract：ObjectiveTo explore the effect of Qingfei Jiangmai decoction （QJD） on the content of mercapturic acids in urine in healthy people amid PM2.5 （particles 2.5 microns or less in size） pollution.MethodA total of 84 healthy students of 18-30 years old in Beijing were recruited and they were randomized into the test group （42 in total， with 1 dropout） and control group （42 in total， with 3 dropouts）. During the pollution， the test group and the control group respectively took QJD granules and placebo for 7 days （1 bag/time， 2 times/day）， and another 7-day intervention with the same drugs was performed at an interval of 4 weeks. The time-activity patterns were recorded during the intervention. On-line solid phase extraction-liquid chromatography/tandem mass spectrometry （SPE-LC-MS/MS） was performed to detect the content of PM2.5-related metabolites S-phenylmercapturic acid （SPMA）， 3-hydroxypropylmercapturic acid （3-HPMA）， 3-hydroxy-1-methylpropylmercapturic acid （HMPMA）， N-acetyl-S-（2-nitrile ethyl）-L-cysteine （CEMA）， and N-acetyl-S-（2-hydroxy ethyl）-L-cysteine （HEMA） in urine before and after intervention. Statistical analysis was followed.ResultThe content of CEMA， HEMA， 3-HPMA， and HMPMA in the test group was all higher after the intervention than before the intervention， with the significant difference in HEMA （P<0.05）. After intervention， content of HEMA and SPMA was significantly higher in the test group than in the control group （P<0.05）， and the difference in HEMA （Z=-3.614， P<0.01） and HMPMA （Z=-1.988， P<0.05） before and after invention in the test group was significantly larger than that in the control group. After the intervention， HEMA in the test group was significantly higher than that in the control group （F=7.597， P<0.01）.ConclusionDuring PM2.5 pollution， QJD can increase the excretion of HEMA， a metabolite of ethylene oxide， in the urine of healthy people in Beijing， and enhance the detoxification process of toxic components in PM2.5， which is of great value in preventing and treating haze-related illnesses.
Keywords：particulate matter 2.5（PM2.5）;mercapturic acids;Qingfei Jiangmai decoction;Chinese medicine therapy;haze;randomized control trial
Abstract：ObjectiveTo observe the efficacy and safety of Fuzheng Huayu tablets （FHT） for treating pulmonary inflammation in patients with coronavirus disease 2019 （COVID-19）.MethodA total of 70（4 cases were lost to follow-up, and 66 cases were finally completed） COVID-19 patients were recruited from February 1 to April 15 in 2020. They were assigned to a control group （35 patients） and a FHT group （31 patients）. The patients in the control group received routine treatment alone and those in the FHT group received FHT in addition to routine treatment. The primary outcome was the ratio of patients showing improvement in chest computed tomographic manifestations after 14 days. The secondary outcome measures included remission rate or progression rate of critical illness， clinical remission rate of respiratory symptoms， routine blood examination， C-reactive protein （CPR） level， procalcitonin （PCT） level， and blood oxygen saturation （SPO2）. The safety was assessed based on liver and kidney functions and adverse events.ResultAfter the 14-day treatment， the ratio of patients showing improvement in the FHT group （100%） was higher than that in the control group （77.1%） （χ2=8.063，P<0.01）. The ratio of disease stages after treatment showed no significant difference between two groups. In the FHT group， the symptoms including cough， dyspnea， and fatigue were alleviated after treatment （P<0.01）. In the control group， the symptoms including fever， cough， and dyspnea were alleviated （P<0.01）， while the fatigue was not relieved after treatment. No significant difference was observed in the clinical symptoms between the two groups after treatment. After treatment， the FHT group showed decreased white blood cell （WBC） count and neutrophil-to-lymphocyte ratio （NLR） （P<0.01）， elevated platelet （PLT） level （P<0.05）， lowered CRP level （P<0.05）， and no significant difference in lymphocyte （LYM）， hemoglobin （Hb）， SPO2 or PCT level. The control group showed decreased NLR （P<0.05） and WBC count （P<0.01）， elevated PCT level （P<0.05）， and no significant change in LYM， Hb， PLT， SPO2 or CRP level after treatment. Furthermore， the FHT group had higher PLT level than the control group （P<0.05） after treatment， and other indicators had no significant differences between the two groups. The liver and kidney functions had no significant difference between the two groups after treatment.ConclusionFHT can safely promote the absorption of acute pulmonary inflammation in COVID-19 patients.
Abstract：ObjectiveTo explore the effect of Ganshuang granule on liver fibrosis （S1 and S2） in chronic hepatitis B （CHB） with liver depression spleen deficiency and blood stasis syndrome.MethodA total of 100 patients were classified into the control group （50 in total with 4 lost and 2 rejected， 44 finally included） and observation group （50 in total with 5 lost and 2 rejected， 43 finally included） with the random number table method. Both groups were given oral entecavir tablets （0.5 mg/time， once a day， 12 months）， and oral glutathione tablets was applied depending on the conditions of patients. In addition， the control group took the analog drug of Ganshuang granule （3 g/time， 3 times/day， 12 months） and the observation group received Ganshuang granules （3 g/time， 3 times/day， 12 months）， followed by histological examination of the liver by puncture biopsy. The two groups were compared in terms of inflammatory activity grade and fibrosis stage， as well as liver stiffness measure （LSM）， liver function， hepatitis B virus （HBV） DNA， liver depression and spleen deficiency syndrome score， aspartate aminotransferase （AST）-to-platelet ratio index （APRI）， and fibrosis index based on the four factors （FIB-4）.ResultAfter treatment， liver fibrosis in the observation group was milder than that in the control group （P<0.05） and the inflammatory activity grade in the observation group was lower than that in the control group （P<0.05）. The effective rate in down-regulating inflammatory activity grade in the observation group was 77.78% as compared with the 45.83% in the control group （χ2=5.546， P<0.05）. The effective rate in decreasing the fibrosis stage in the observation group was 59.26%， which was higher than that （16.67%） in the control group （χ2=9.669， P<0.01）. The LSM and score of the liver stagnation and spleen deficiency syndrome in the observation group were lower than those in the control group at the 6th months and 12th months of treatment （P<0.05，P<0.01）. The levels of alanine aminotransferase （ALT）， AST， and alkaline phosphatase （ALP） in the observation group were lower than those in the control group （P<0.01）. The APRI and FIB-4 in the observation group were lower than those in the control group （P<0.01）.ConclusionThe Ganshuang granule combined with entecavir can alleviate inflammation and liver fibrosis， delay and reverse liver fibrosis， protect liver， and improve the traditional Chinese medicine syndrome of liver fibrosis （S1 and S2） in CHB， which is worth of clinical use and further research.
Keywords：chronic hepatitis B;liver fibrosis;liver depression and spleen deficiency syndrome;Ganshuang granules;early prevention and treatment;degree of fibrosis
Abstract：ObjectiveTo observe the clinical efficacy and safety of Chinese medicinal mixture for dispelling stasis and resolving phlegm combined with western medicine in the treatment of epilepsy combined with cognitive impairment by randomized， double-blind， placebo-controlled， parallel-group trial.MethodA total of 123 inpatients and outpatients with epilepsy complicated with cognitive impairment admitted to the department of neurology at Longhua Hospital from October 2020 to October 2021 were randomly assigned into a control group （62 cases， carbamazepine + placebo） and a treatment group （61 cases， carbamazepine + Quyu Dingxian Zhengtong mixture） by random number table method. In the treatment group， 4 cases were exfoliated and eliminated. In the control group， 3 cases fell off. Finally， 57 cases in the treatment group and 59 cases in the control group were included. The total course of treatment for both groups was 12 weeks. The clinical efficacy， efficacy for traditional Chinese medicine （TCM） syndromes， and incidence of adverse reactions were compared between two groups. The changes of seizure frequency， abnormal rate of electroencephalogram （EEG）， cognitive function score， serum homocysteine （HCY）， folic acid， and vitamin B12 （B12） were measured and compared before and after treatment.ResultAfter 12 weeks of treatment， the treatment group had higher clinical efficacy ［92.98% （53/57） vs 79.66% （47/59）， χ2=4.327， P<0.05］ and efficacy for TCM syndromes ［96.49% （55/57） vs 84.75% （50/59）， χ2=4.660， P<0.05］ than the control group. The treatment group was superior to the control group in reducing the seizure frequency （Z=-3.938， P<0.01） and improving the Montreal cognitive assessment （MoCA） score （t=4.333， P<0.01） and mini-mental state examination （MMSE） score （t=9.531， P<0.01）. The variations in serum HCY， folic acid， and B12 in the treatment group were less than those in the control group （t=-7.233， t=-7.972， t=-6.871， P<0.01）. After treatment， the abnormal rate of EEG in the treatment group was lower than that in the control group （χ2=4.437， P<0.05）. The incidence of adverse reactions in the treatment group （1.75%， 1/57） was lower than that （13.56%， 8/59） in the control group （corrected χ2=4.116， P<0.05）.ConclusionChinese medicinal mixture for dispelling stasis and resolving phlegm in combination with western medicine had better efficacy and safety than western medicine alone in the treatment of epilepsy complicated with cognitive impairment. Specifically， the combination outperformed western medicine alone in terms of clinical efficacy， efficacy for TCM syndromes， reduction in seizure frequency， abnormal rate of EEG， adverse reactions， improvement of cognitive function， and variations in serum folic acid， B12， and HCY values. Chinese medicinal mixture for dispelling stasis and resolving phlegm may improve the clinical efficacy and safety by changing the metabolism of folic acid， B12， and HCY in serum of the patients with epilepsy complicated with cognitive impairment.
Keywords：dispelling stasis and resolving phlegm;epilepsy with cognitive impairment;randomized double-blind parallel-group trial;clinical research;efficacy
Abstract：ObjectiveTo explore the effect of Qinggan Zishen prescription on metabolic disorders in obesity-related hypertension （OBH） patients and analyze the potential pharmacological mechanism based on network pharmacology.MethodA total of 85 eligible OBH patients who were treated in the outpatient or wards of Jiangsu Province Hospital of Chinese medicine from September 2018 to January 2020 were selected and randomized into the observation group （45 cases） and control group （40 cases）. All patients were treated with western medicine during a four-week introduction period， and then the observation group was treated with Qinggan Zishen prescription on the basis of western medicine. The study lasted 6 months， and indicators， such as triglycerides （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， glycosylated hemoglobin （HbA1c）， fasting blood glucose （FBG）， fasting insulin （FINS）， waist circumference （W）， hip circumference （H） were detected and homeostasis model assessment of insulin resistance （HOMA-IR），body mass index （BMI）， waist-hip ratio （WHR） were calculated before and after intervention. At the same time， the regulation network of the Qinggan Zishen prescription was visualized and the protein-protein interaction （PPI） network was constructed. The core targets of the network were obtained for Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis.ResultAfter intervention for 6 months， the levels of W， H， WHR， FINS， and HOMA-IR in the observation group were reduced as compared with those in the control group （P<0.05， P<0.01）. According to network pharmacology， the main components of Qinggan Zishen prescription in treating OBH were luteolin， quercetin， and berberine and the key targets were amyloid precursor protein （APP）， vascular endothelial growth factor A （VEGFA）， and estrogen receptor 1 （ESR1）. Moreover， the key biological pathway was advanced glycation end product （AGE）/advanced glycation end product receptor （RAGE） signaling pathway.ConclusionQinggan Zishen prescription can improve the metabolic disorder of OBH patients through multiple components， multiple targets， and multiple pathways， which provides new mindset for follow-up studies.
Abstract：ObjectiveTo optimize the extraction and purification process of Gardeniae Fructus for industrial production， and to obtain the total iridoid and total crocin extracts.MethodOrthogonal test was used to optimize the water extraction process by taking contents of geniposide， genipin gentiobioside， gardenoside， crocin-1 and crocin-2 as indicators and the decocting time， decocting times and water amount as factors. The purification process was optimized by single factor test， and four different types of macroporous adsorption resins were screened. The process conditions such as resin type， maximum loading amount， water washing amount， ethanol concentration， ethanol dosage， and flow rate of sample loading were mainly investigated. In addition， the drying methods （vacuum drying and spray drying） of the extract were investigated， and a pilot scale-up verification test was carried out.ResultThe optimal water extraction process of Gardeniae Fructus was to add 15， 10 times the amount of water for decocting twice， 1 h each time. The optimal purification process was as follows：the water extract through SP825L macroporous resin column， the amount of crude drug-the amount of resin （1∶1.5）， the sample loading flow rate of 3 BV h-1， adding 2 BV of water to remove impurities， adding 4 BV of 30% ethanol to obtain the iridoid part， then adding 3 BV of 70% ethanol to obtain the crocin part， collecting the ethanol lotion， and drying at 70 ℃. Under these conditions， the extraction amount of total iridoids was 590.75 mg·g-1 with the transfer rate of 70.48%， and the yield of dry extract was 8.89%. The extraction amount of total crocins was 83.37 mg·g-1 with the transfer rate of 22.20%， and the dry extract yield was 2.60%.ConclusionThe optimized extraction and purification process is stable and feasible with high extraction rate of active components， which is suitable for the industrial extraction and purification of active parts of Gardeniae Fructus.
Keywords：Gardeniae Fructus;macroporous resin;iridoid;crocin;orthogonal test;ultra-high performance liquid chromatography （UPLC）;purification process
Abstract：ObjectiveTo explore the correlation between the content of 4 functional components in Codonopsis pilosula roots from different areas and soil factors， and thereby to lay a theoretical basis for soil ecological regulation and improvement of quality of C. pilosula roots.MethodThe content of lobetyolin， atractylenolide Ⅲ， alcohol extract， and polysaccharides， as well as soil fertility and 16 soil factors in 24 batches of samples from different producing areas were determined. Principal component analysis （PCA） and Pearson's correlation analysis were used to explore the key soil factors leading to the variation of chemical component content in C. pilosula roots.ResultThe content of lobetyolin and atractylenolide Ⅲ in samples from Longxi was the highest， and the content of polysaccharides peaked in samples from Huguan. The content of lobetyolin was in positive correlation with soil total nitrogen， total phosphorus， total potassium， alkali-hydrolyzable nitrogen， and available potassium （P<0.01）， as well as soil organic matter， pH， available manganese， and available zinc （P<0.05）. There was a positive correlation between pH and atractylenolide Ⅲ content （P<0.05）. Soil total potassium was in positive correlation with alcohol extract and polysaccharide content （P<0.01）. Soil available zinc was positively correlated with alcohol extract and the polysaccharide content （P<0.05）. Sample sites with higher PCA scores were Pingshun， Huguan， and Longxi， which were significantly positively correlated with the content of polysaccharides in C. pilosula roots in different habitats.ConclusionThe content of functional components in C. pilosula roots can be improved by raising soil organic matter content and applying specific fertilizers.
Keywords：Codonopsis pilosula roots;functional component;polysaccharides in Codonopsis pilosula roots;soil factor;correlation analysis
Abstract：ObjectiveTo explore the possible mechanism of dried fruiting bodies of Fomes officinalis （FOA） against Alzheimer's disease （AD） based on network pharmacology and experimental verification.MethodThe effective components of FOA were retrieved from a Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine （BATMAN-TCM） and previous reports. The targets of the components were searched from PharmMapper and TargetNet， and the targets related to AD from Gene Expression Omnibus （GEO）， DrugBank， among other databases. Thereby， the common targets of FOA and AD were obtained， and the protein-protein interaction （PPI） network and component-target network were established based on STRING and Cytoscape 3.7.1， followed by the topology analysis of the networks， and Gene Ontology （GO） term enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis of the common targets. The results were verified by the molecular docking and the in vitro cell experiment.ResultA total of 24 candidate components and 242 predicted targets of FOA， and 96 common targets of FOA and AD were screened out. The key components included ［2-（1-carboxyhexadecylamino）-2-aminosuccinic acid］， 3-keto-dehydrosulfurenic acid， and eburicoic acid， and the active targets were albumin （ALB）， acetylcholinesterase （AChE）， estrogen receptor 1 （ESR1）， cysteine aspartate-specific protease-3 （Caspase-3）， and beta-secretase1 （BACE1）. The common targets were involved in 392 GO terms， and the key terms were the β-amyloid metabolic process and cholinesterase activity. A total of 77 KEGG pathways were obtained， which mainly included estrogen signaling pathway， cholinergic synapse， and AD. The results of molecular docking showed that 7 components of FOA had high binding affinity to amyloid precursor protein （APP）， BACE1， AChE， and Caspase-3. The cell survival rate rose （P<0.01） and the mRNA and protein expression of APP， BACE1， AChE， and Caspase-3 reduced in FOA groups in a dose-dependent manner compared with those in the model group （P<0.05）.ConclusionThis study reveals for the first time that FOA has multi-component， multi-target， and multi-pathway characteristics in the treatment of AD， which serves as a reference for further explaining the mechanism of FOA against AD.
Keywords：dried fruiting bodies of Fomes officinalis;Alzheimer's disease （AD）;human neuroblastoma （SHSY5Y） cell;network pharmacology;mechanism of action
Abstract：ObjectiveTo explore the mechanism of Cordyceps in treating bronchial asthma and chronic renal failure with the concept of "same treatment for different diseases" in traditional Chinese medicine （TCM） by network pharmacology and molecular docking technology.MethodThe active components and potential targets of Cordyceps were collected from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP） and SwissTargetPrediction. The disease targets were obtained from Therapeutic Target Database （TTD）， DrugBank， GeneCards and other databases. The common targets were obtained from the intersection of potential targets and disease targets. The protein-protein interaction （PPI） network was constructed by STRING11.5， and the ''component-target-diseas'' network of Cordyceps was established by Cytoscape 3.9.0. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses were carried out by Metascape， and molecular docking was performed by Autodock 4.2.ResultSixty common targets of disease and drug were screened out. The core targets mainly involved protein kinase B1 （Akt1）， non-receptor tyrosine kinase， sarcoma virus protein （SRC）， TP53， matrix metalloproteinase-9 （MMP-9）， and prostaglandin endoperoxide synthase 2 （PTGS2）. The potential targets were mainly enriched in the signaling pathways of renin-angiotensin system （RAS）， RAP1， phosphoinositide 3 kinase/protein kinase B （PI3K/Akt）， mitogen-activated protein kinase （MAPK）， etc.ConclusionThe active components of Cordyceps inhibited inflammatory response and reduced fibrosis and cell apoptosis in a multi-target and multi-pathway manner. The findings of this study preliminarily revealed the potential targets and modern biological mechanism of Cordyceps in treating bronchial asthma and chronic renal failure with the concept of ''same treatment for different diseases''， and provided references for in-depth experimental verification and clinical application.
Keywords：Cordyceps;network pharmacology;molecular docking technology;bronchial asthma;chronic renal failure;same treatment for different diseases
Abstract：In the "major preventable diseases and health problems in China in the next 20 years" released by the Chinese Academy of Engineering， hypertension takes the second place. As a metabolic disease， hypertension has become a major global public health problem. In recent years， owing to the multiple targets， multiple components， and holistic regulation， traditional Chinese medicine （TCM） has attracted the interest of scholars in the treatment of hypertension. Especially for young patients and patients with early-onset hypertension， it is expected to achieve the purpose of stopping or reducing western medicine and decreasing adverse reactions by syndrome differentiation and treatment. Through literature review and expert consultation， the Society of Cardiovascular Diseases of China Association of Chinese Medicine has organized experts to formulate comprehensive intervention strategies of TCM for hypertension and finally developed the Expert Consensus on Diagnosis and Treatment of Hypertension with Traditional Chinese Medicine （hereinafter referred to as the Consensus）. It was demonstrated and approved at the annual meeting of the Society of Cardiovascular Diseases of China Association of Chinese Medicine in December 2018 and officially published in August 2019. Combined with literature research and clinical antihypertensive practice of TCM， this paper aims to make a detailed interpretation of the key pathogenesis， syndromes， classical prescriptions， and syndrome differentiation-based medication in the Consensus， which is expected to promote the clinical popularization and application of the Consensus. The conclusions are as follows. The pathogenesis of hypertension can be classified into fire syndrome， fluid-retention syndrome， and deficiency syndrome. In terms of syndrome characteristics， hypertension is mainly divided into three major syndromes： ascendant hyperactivity of liver Yang， stagnation of phlegm-fluid， and deficiency of kidney Yin. Moreover， it details the key points in the treatment of hypertension with the recommended classical prescriptions and the medication rules. As for the non-drug therapy， Taiji， Baduanjin， Qigong， yoga， acupuncture， moxibustion， massage， pricking collaterals， cupping， and foot bathing are also recommended to relieve clinical symptoms and control blood pressure. In terms of health care， it is emphasized to pay attention to "uncontrollable factors of blood pressure" such as exogenous factors， emotional disorders， insomnia， and constipation.
Keywords：hypertension;traditional Chinese medicine;expert consensus;interpretation
Abstract：The incidence of psoriasis which is characterized by dryness， scaling and itching of the skin has been on the rise. It can have a profound psychological impact on patients' quality of life. Accumulating research has been conducted on the mechanisms of psoriasis in western medicine， from the difference of pathological manifestations of terminal keratinocytes， the disorder of expression of related factors and cells， to the immune imbalance of T lymphocytes and their subsets， or the abnormal transcription dominated by genetic genes. As a result， a complex and huge mechanism network has formed and many hypotheses have emerged. In recent years， with the in-depth research on non-coding genes， it has been clarified that microRNA-modified multi-pathway effect interferes with the occurrence and development of psoriasis and affects the proliferation， apoptosis， and differentiation of keratinocytes. The research on microRNA involves both genetics and immunology， which can help improve the key links in the micro pathway of psoriasis. Thus， it is a key part in the pathogenesis of psoriasis and has also become the hotspot and difficulty of modern research on psoriasis. At the same time， we should give full play to the advantages of traditional Chinese medicine （TCM） in syndrome differentiation and treatment. To be specific， microRNA targets of compound Chinese medicine preparations with the functions of clearing heat， cooling blood， detoxifying and removing blood stasis or effective medicinal monomers such as paeonol， tripterygium glycosides， shikonin， curcumin， total glucosides of paeony and indirubin should be explored， and microRNA can be used as the basis for blood syndrome differentiation of psoriasis. Thereby， the syndrome differentiation theory of TCM and micro indicators of western medicine are integrated， to make full use of characteristics of TCM and guide the clinical syndrome differentiation and treatment. At present， the intervention on microRNA in TCM is rarely studied， and available studies mainly focus on several targets such as microRNA-155， microRNA-210， microRNA-21， microRNA-203， microRNA-320， microRNA-124， microRNA-330， microRNA-146a， and microRNA-15a-5p. This paper summarizes the research on compound Chinese medicine prescriptions and monomers in the treatment and syndrome differentiation of psoriasis through the intervention of microRNA， which is expected to provide a reference for the research on psoriasis in TCM and western medicine and the establishment of microRNA-based database.
Keywords：traditional Chinese medicine;microRNA;psoriasis;mechanism;classification
Abstract：Smallanthus sonchifolius， a plant resource with both medicinal and edible values， has been taken as fruit for a long history. Studies have proved that phenolic acids， flavonoids， sesquiterpene lactones， and fructooligosaccharides are the major compounds in S. sonchifolius. The extract of S. sonchifolius demonstrates noticeable antioxidant， anti-inflammatory， antimicrobial， and anti-cancer effects， as well as the activities of lowering blood glucose level， regulating intestinal function and so on. The rhizomes and leaves of S. sonchifolius contain abundant phenolic acids， mainly caffeic acid and its derivatives， which endow S. sonchifolius with remarkable antioxidant effect. Moreover， these substances can reduce blood glucose by improving insulin sensitivity. Fructooligosaccharides are abundant in the tuber of this plant， which can improve intestinal function by regulating intestinal flora. The sesquiterpene lactones in glandular trichomes on the leaf surface can inhibit the proliferation of cancer cells， among which uvedafolin and enhydrofolin have particularly strong activities. Furthermore， the sesquiterpene lactones have obvious inhibitory effect on Gram-positive bacteria. In terms of structure， the number of epoxy groups is linked to the strength of anticancer and antimicrobial effects. In addition， S. sonchifolius contains other compounds such as volatile oils， fatty acids， sterols， diterpenes， p-hydroxyacetophenone derivatives， and octulosonic acid derivatives， thereby exhibiting the pharmacological effects of treating Alzheimer's disease， protecting kidney， and lowering blood lipids. However， the isolation and identification of the main compounds in S. sonchifolius need further exploration， and the mechanism of action remains to be studied. Here we summarized the principal chemical components and pharmacological activities of S. sonchifolius， aiming to give a clue for the comprehensive development and utilization of this plant.
Abstract：Ferroptosis， a new type of iron-dependent programmed cell death， is related to multiple pathways such as glutathione/glutathione peroxidase 4， iron metabolism， lipid metabolism， and iron autophagy， and plays an important part in the occurrence and development of many diseases， such as tumor， cerebral ischemia， and Parkinson's disease. Ferroptosis is a double-edged sword as it can eliminate pathological cells （such as tumor cells） but long-term ferroptosis may cause or aggravate other disorders related to abnormal lipid metabolism and iron metabolism. Regulating the balance between cell proliferation and ferroptosis may be an important target for drug intervention in diseases. The Yin-yang theory is one of the foundational principles of traditional Chinese medicine （TCM）， which is used to explain the physiological functions and pathological changes of human body and to guide the diagnosis and prevention of disease and health care. The balance of cell proliferation and programmed death is essentially the balance of Yin and Yang at the cellular level， which is governed and regulated by the law of balance. TCM intervenes in ferroptosis by promoting ferroptosis of tumor cells （damaging the excess） and inhibiting ferroptosis of other diseases （compensating the deficiency）， which is similar to the treatment principle of adjusting Yin and Yang. On this basis， this article aims to use the Yin-yang theory to clarify the relationship between TCM promoting ferroptosis and inhibiting ferroptosis， which is expected to lay a basis for the modern application of Yin-yang theory and provide new targets for TCM treatment.
Keywords：ferroptosis;traditional Chinese medicine;Yin-yang theory;damaging the excess;compensating the deficiency
Abstract：As a traditional Chinese medicinal material， Glycyrrhizae Radix et Rhizoma has been extensively used in various formulae. According to modern pharmacological research， it has anti-tumor， anti-inflammatory， anti-viral， liver-protecting， anti-heart failure， immunoregulatory， and anti-fibrosis effects. Caused by the interaction of various factors， cancer features complex pathogenesis. It is a global challenge and one of the main causes of death in China. Statistics show that the morbidity and mortality of malignant tumors have been on the rise， particularly for the young， which threaten the health of human beings. At the moment， radiotherapy and chemotherapy are the main countermeasures. Most clinical anti-tumor drugs demonstrate non-selective toxicity. To be specific， they damage normal cells while killing tumor cells， thus injuring vital organs. In addition， long-term medication will reduce the sensitivity of tumor cells. However， traditional Chinese medicine， which is characterized by treatment based on syndrome differentiation， multiple components， and multiple targets， is superior in the treatment of tumor. Studies have shown that the combination of anti-tumor drugs with Chinese medicine can not only enhance the anti-tumor effect but also alleviate toxicity. Therefore， it has been a research hotspot to develop anti-tumor drugs based on traditional Chinese medicine. In recent years， major headway has been made in the research on active ingreddients of Glycyrrhizae Radix et Rhizoma in anti-liver cancer， anti-breast cancer， anti-lung cancer， and anti-colon cancer and the combination with other drugs for anti-tumor. On this basis， we summarized the mechanisms of active ingredients of Glycyrrhizae Radix et Rhizoma in inducing apoptosis， interfering with cell cycle， inducing autophagy， inhibiting glycolysis， regulating immunity， and modulating miRNA and signaling pathways， as well as the combination with other drugs in anti-tumor efficiency， toxicity reduction， and sensitivity enhancement， hoping to lay a theoretical basis for the further development and clinical application of active ingredients of Glycyrrhizae Radix et Rhizoma.
Keywords：Glycyrrhizae Radix et Rhizoma;active ingredients of Glycyrrhizae Radix et Rhizoma;anti-tumor;mechanism;combined anti-tumor
Abstract：Autophagy and tumor immune escape are important biological mechanisms in the process of tumor cell proliferation and metastasis， involving multiple signaling pathways. The interaction of autophagy and tumor immune escape seriously affects the treatment and prognosis of tumor diseases. However， the correlation between autophagy and tumor immune escape is still not fully elucidated. Recent studies have shown that autophagy can affect the activity of immune cells by regulating the presentation of antigens in tumor cells， the release of cytokines， and the degradation of immune checkpoint proteins， thereby positively or negatively regulating tumor cell immune escape. The activation of autophagy in tumor cells can inhibit the activation of the innate immune sensing pathway of stimulator of interferon genes （STING）-type Ⅰ interferon （IFN-Ⅰ） to inhibit its immunogenicity and cytotoxic T lymphocytes （CTLs）， which promotes tumor immune escape. While autophagy suppression can reduce the infiltration of M2 macrophages， promote the binding of natural killer group 2， member D （NKG2D） to its ligand， and inhibit the recognition of immune checkpoint proteins， thereby exerting an immune-killing effect and inhibiting tumor immune escape. Traditional Chinese medicine （TCM） has unique advantages in anti-tumor research， especially in the unilateral regulation of autophagy or improvement of tumor immunity， but the research based on the regulation of autophagy and tumor immunity by TCM is insufficient. A few studies have shown that Chinese medicine monomers and compounds can exert an anti-tumor effect by regulating cell autophagy and interfering with tumor immune escape， but there is still a lack of systematic elaboration. The present study reviewed correlation between autophagy and tumor immune escape and regulation of autophagy by Chinese medicine to interfere with tumor immune escape to provide new ideas for research on mechanism of TCM against tumor diseases and development of innovative TCM drugs against tumors.
Abstract：Gouty arthritis （GA） is the metabolic rheumatism caused by purine metabolism disorder， which can be acute or chronic. The main manifestations of GA include recurrent redness， swelling， heat pain， and dysfunction of the affected joints. According to the theory of modern medicine， GA is closely associated with the increase in uric acid， the participation of inflammatory cytokines， the weakening of antioxidant response， apoptosis， and the imbalance of intestinal flora and bone metabolism， whereas the specific pathogenesis remains unclear. GA is characterized by easy diagnosis， difficult treatment， and high recurrence rate， which seriously affects the life quality of patients. Colchicine， corticosteroids， non-steroidal anti-inflammatory drugs， and selective cyclooxygenase-2 inhibitors are the commonly used western medicines for this disease， which demonstrate remarkable short-term therapeutic effect. However， long-term use of these medicines will bring serious adverse reactions. Chinese medicines， with high safety and causing few adverse reactions， have a variety of active components which can act on multiple pathways and targets to exert synergistic effects， thus attracting wide attention. This paper systematically reviews the literature reporting the Chinese medicines in improving antioxidant response， reducing chondrocyte apoptosis， and regulating intestinal flora and bone metabolism， aiming to further clarify the pathogenesis of GA and provide a scientific basis for the clinical application of Chinese medicines in the prevention and treatment of GA.
Keywords：gouty arthritis;traditional Chinese medicine;pathogenesis;monosodium urate;research progress
Abstract：Fibrosis can occur in nearly all organs of the body and is an outcome of many chronic diseases. As inflammation leads to necrosis of parenchymal cells， excessive proliferation of fibroblasts and overproduction of extracellular matrix （ECM） occur in tissues and organs， which may cause structural damage and loss of function of organs in the case of continuous progression. Chinese medicine has definite effect on fibrosis and prescriptions with effects of replenishing Qi and activating blood， such as Buyang Huanwutang， are frequently used in clinical settings. Clinical research and experiments show that Buyang Huanwutang can delay the progression of fibrosis in multiple organs such as lung， heart， liver， and kidney by improving organ function， reducing ECM deposition， anti-oxidative stress， anti-inflammatory response， regulating the imbalance of matrix metalloproteinases （MMPs）/tissue inhibitors of metalloproteinases （TIMPs）， and modulating transforming growth factor-β （TGF-β）/Smad pathway. According to traditional Chinese medicine， healthy Qi deficiency is the internal cause of fibrosis， and blood stasis is an important pathological factor in the formation of fibrosis. Moreover， deficiency and stasis exist in the whole process of fibrosis and the changes of microenvironment of fibrotic organs and tissues accord with the pathological manifestations of Qi deficiency and blood stasis. This article reviews the anti-fibrosis mechanism of Buyang Huanwutang in multiple organs， which provides a science-based explanation for the treatment of fibrosis by Buyang Huanwutang and lays a foundation for further clinical research.
Keywords：Buyang Huanwutang;organ fibrosis;mechanism of action;research progress;inflammation
Abstract：Diabetic nephropathy （DN） is among the common microvascular complications of diabetes. In recent years， the incidence has been on the rise with the increase in prevalence of diabetes， threatening the health of human. The early stage of DN is characterized by excessive accumulation of extracellular matrix （ECM） and thickening of glomerular basement membrane which result in glomerular mesangial proliferation and massive collagen deposition. The late stage features glomerular sclerosis and renal fibrosis （RF）. It has been confirmed that RF is the key pathological process for the development of DN. Therefore， it is the research focus to explore the pathogenesis and treatment methods of RF. It has been frequently verified that Chinese medicine is superior in the treatment of diabetic RF. It relieves diabetic RF by regulating transforming growth factor-β （TGF-β）， secretory glycoprotein （Wnt）/β-catenin， nuclear factor-κB （NF-κB）， Notch， mitogen-activated protein kinase （MAPK）， and other signaling pathways. Therefore， this paper reviews the pathogenesis of diabetic RF and the treatment with Chinese medicine， which is expected to serve as a reference for clinical application of Chinese medicine in the treatment of diabetic RF.
Keywords：diabetic nephropathy;renal fibrosis;pathogenesis;traditional Chinese medicine;Chinese medicinal materials