Abstract：ObjectiveTo investigate the neuroprotective effect of Danggui Shaoyaosan （DSS） in a rat model of amyloid-β-peptide1-42 （Aβ1-42）-induced Alzheimer's disease （AD） as well as its regulatory effect on NOD-like receptor protein 3 （NLRP3）/cysteinyl aspartate-specific protease-1 （Caspase-1） signaling pathway.MethodThe AD animal model was established via intracerebral injection of Aβ1-42 and treated with different concentrations of DSS after the division of rats into the sham operation group， model group， as well as the high-， medium-， and low-dose DSS groups. Morris water maze test was conducted to determine the learning and memory abilities of rats. The morphology and function of neurons were detected by hematoxylin-eosin （HE） staining and Golgi staining， followed by immunofluorescence co-localization of NLRP3 inflammasome activation. The mRNA expression levels of interleukin （IL）-1β and IL-18 were measured by Real-time polymerase chain reaction （Real-time PCR）， and the protein expression levels of NLRP3， Caspase-1， and IL-1β were assayed by Western blot.ResultCompared with the sham operation group， the model group exhibited significantly decreased learning and memory abilities （P<0.01）， impaired neuronal morphology and function， up-regulated IL-1β and IL-18 mRNA expression， enhanced NLRP3 inflammasome activation， and elevated NLRP3， Caspase-1， and IL-1β protein expression （P<0.01）. Compared with the model group， DSS at both medium and high doses remarkably improved the learning and memory abilities of AD rats （P<0.05， P<0.01）， restored neuronal morphology and function， down-regulated the mRNA expression levels of inflammatory factors IL-1β and IL-18， reduced the activation of NLRP3 inflammasomes， and lowered the protein expression levels of NLRP3， Caspase-1， and IL-1β （P<0.01）.ConclusionDSS inhibits inflammasome activation and neuroinflammatory response possibly by regulating the NLRP3/Caspase-1 signaling pathway， thus exerting the neuroprotective effect.
Abstract：ObjectiveTo explore the protective effect and the mechanism of Danggui Shaoyaosan（DSS） on angiotensin Ⅱ （AngⅡ）/transient receptor potential cation channel 6 （TRPC6） pathway in nephrotic syndrome （NS） rats.MethodIn animal experiments， doxorubicin （4 mg·kg-1 for the 1st week and 2 mg·kg-1 for the 2nd week） was injected twice to the tail vein of rats to induce NS model in 160 rats， which were then randomly divided into model group （normal saline）， losartan group （30 mg·kg-1·d-1）， and low-（4.3 g·kg-1·d-1）， medium-（8.6 g·kg-1·d-1）， and high-dose （17.2 g·kg-1·d-1） DSS groups. Besides， a normal group was also set. After intervention for four weeks， ultrastructure changes of the kidney were identified by transmission electron microscopy （TEM）. The 24-hour urine protein was detected by kits. Radioimmunoassay was used to detect the content of AngⅡ and Calcineurin （CaN） in plasma. Western blot was used to detect the protein expression of TRPC6， angiotensin Ⅱ type 1 receptor （AT1R）， podocyte slit diaphragm-specific protein （Nephrin）， and cysteine-aspartic acid protease-3 （Caspase-3） in the renal cortex. Immunohistochemistry was used to detect the expression of TRPC6 and AT1R in the slit diaphragm. In cell experiments， AngⅡ stimulated MPC5 podocytes. The cells were randomly divided into a normal group， an AngⅡ group， an AngⅡ+SAR7334 （TRPC6-specific inhibitor） group， an AngⅡ+5%DSS group， an AngⅡ+10%DSS group， and an AngⅡ+15%DSS group. Western blot was used to detect the protein expression of TRPC6， AT1R， Nephrin， and Caspase-3 in podocytes.ResultCompared with the normal group， the model group showed increased 24-hour urine protein content （P<0.01） and AngⅡ and CaN in plasma （P<0.01）， incomplete glomerular structure， the extensive fusion of podocyte process with elevated fusion rate （P<0.01）， increased expression distribution of AT1R and TRPC6 in the renal cortex， and up-regulated protein expression of AT1R， TRPC6， and Caspase-3 in renal tissues （P<0.01）， and reduced Nephrin protein expression （P<0.01）. Compared with model group， the losartan group and the high-dose DSS group exhibited decreased 24-hour urine protein content （P<0.01） and the content of AngⅡ and CaN in plasma （P<0.01）， improved glomerular structure， reduced fusion rate of podocyte process （P<0.01）， diminished expression distribution of TRPC6 and AT1R in the renal cortex， declining protein expression of AT1R， TRPC6 and Caspase-3 in renal tissues （P<0.01）， and elevated Nephrin protein expression （P<0.01）. Additionally， compared with the normal podocytes， AngⅡ-stimulated podocytes showed increased protein expression of AT1R， TRPC6 and Caspase-3 （P<0.01）， and decreased expression of Nephrin （P<0.01）. Compared with the AngⅡ group， the AngⅡ+SAR7334 group displayed reduced protein expression of AT1R， TRPC6， and Caspase-3 （P<0.01） and increased protein expression of Nephrin （P<0.01）.ConclusionDSS can improve the pathological characteristics of NS presumedly by inhibiting the interaction between AngⅡ and TRPC6 in podocytes and improving the structural integrity of podocytes to repair the damage of glomerular molecular barrier and slow down the progression of NS-induced proteinuria.
Abstract：ObjectiveTo investigate the effect of Xiaoyaosan on depressive behavioral phenotype in mice with vascular dementia （VaD） mice and its possible mechanism.MethodSixty three-month-old male C57/BL6 mice were divided into the normal control group， model group， positive control group， as well as low-， medium-， and high-dose Xiaoyaosan groups. Mice in all groups except for the normal control group underwent bilateral carotid artery stenosis. Two weeks later， they were subjected to chronic restraint stress， 6 h per day， for inducing VaD complicated with depression. Mice in the low-， medium-， and high-dose Xiaoyaosan groups were treatment with intragastric administration of Xiaoyaosan decoction （5， 10， 20 g·kg-1）， the ones in the positive control group with fluoxetine （10 mg·kg-1）， and those in the normal control group and model group with an equal volume of normal saline for four weeks， during which the restraint stress was maintained. The depressive behavioral phenotype of mice was observed in sugar water preference test and tail suspension test. The fluorescence expression of myelin basic protein （MBP） in ventral hippocampus （vHIP） was detected by fluorescence immunoassay. The ultrastructure of myelin sheath in vHIP was observed by transmission electron microscopy. The protein expression levels of MBP， myelin oligodendrocyte glycoprotein （MOG）， myelin-associated glycoprotein （MAG）， triggering receptor expressed on myeloid cells-2 （TREM2）， inducible nitric oxide synthase （iNOS）， arginase 1 （Arg1）， interleukin-Iβ （IL-1β）， tumor necrosis factor-α （TNF-α）， interleukin-4 （IL-4）， and interleukin-10 （IL-10） were assayed by Western blot.ResultAs revealed by behavioral test， compared with the normal control group， the model group exhibited prolonged immobility time and decreased percentage of sugar water preference （P<0.01）. Compared with the model group， Xiaoyaosan significantly shortened the immobility time of mice （P<0.05） and increased the percentage of sugar water preference （P<0.01）. Western blot results showed that the protein expression levels of MBP， MOG， and MAG in vHIP of the model group were remarkably decreased as compared with those of the normal control group （P<0.01）. The protein expression levels of MBP， MOG， and MAG in vHIP of the low-dose Xiaoyaosan group were increased in contrast to those in the model group （P<0.05， P<0.01）， while the protein expression of iNOS was decreased （P<0.01）. The protein expression levels of MBP， MOG， MAG， TREM2， Arg1， IL-4， and IL-10 in the medium- and high-dose Xiaoyaosan groups were up-regulated （P<0.05， P<0.01）， whereas those of iNOS， IL-1β， and TNF-α were down-regulated （P<0.01）. The immunofluorescence findings demonstrated that the mean fluorescence intensity of MBP in the model group declined in comparison with that in the normal control group （P<0.01）， while the mean fluorescence intensities of MBP in the low-， medium-， and high-dose Xiaoyaosan groups were enhanced to different degrees （P<0.01）. It was observed under the transmission electron microscope that the myelin structure of the model group was loosened and the dense layer was separated and irregularly arranged. Xiaoyaosan improved the structural integrity of myelin sheath and the looseness of lamellar structure.ConclusionXiaoyaosan ameliorates the depressive behavioral phenotype of VaD mice， which may be related to the up-regulation of TREM2， the induction of M2 polarization of microglia cells， the enhancement of their anti-inflammatory and phagocytic abilities， and the promotion of damaged myelin sheath regeneration.
Abstract：ObjectiveTo observe the effects of Scutellariae Radix-Hedyotidis Herba on the proliferation of human lung adenocarcinoma A549 cells and the expression of interleukin-6（IL-6）， phosphatidylinositol 3-kinase （PI3K），protein kinase B （Akt）， p-protein kinase B （p-Akt）， mechanistic target of rapamycin （mTOR）， hypoxia-inducible factor-1α （HIF-1α）， and Cyclin D1 at the cellular level， and to explore their molecular mechanism.MethodFollowing the set-up of the blank group （complete medium）， low-， moderate-，and high-dose （20， 40， and 60 mg·L-1） Scutellariae Radix-Hedyotidis Herba groups， and low-， moderate-， and high-dose （5， 10， and 20 mg·L-1） cisplatin groups， the cell were treated with the corresponding drugs for 24， 48， and 72 h for detecting their viability by tetrazolium bromide （MTT） colorimetry. A549 cells were then divided into the blank group， Scutellariae Radix-Hedyotidis Herba group， cisplatin group， and combined medication group and intervened with thecomplete medium， 40 mg·L-1 Scutellariae Radix-Hedyotidis Herba， 10 mg·L-1 cisplatin， and 40 mg·L-1 Scutellariae Radix-Hedyotidis Herba + 10 mg·L-1 cisplatin， respectively， for 24， 48 and 72 h， followed by the measurement of inhibitory effects against the proliferation of A549 cells in each experimental group. The level of IL-6 in cell culture supernatant was determined by enzyme-linked immunosorbent assay （ELISA） after 72 h. The mRNA expression levels of HIF-1α and Cyclin D1 in each group were assayed by real-time polymerase chain reaction （Real-time PCR）， and the protein expression levels of PI3K， Akt， p-Akt， mTOR， HIF-1α， and Cyclin D1 by Western blot.ResultAfter 24 h intervention， Scutellariae Radix-Hedyotidis Herba did not significantly inhibit the proliferation of A549 cells. However， 48 h later， the inhibitory effect in Scutellariae Radix-Hedyotidis Herba groups were significantly enhanced in comparison with that in the blank group （P<0.05）， exhibiting a time-dependent response. After 72 h of action， no significant change was present in the inhibitory effect of each Scutellariae Radix-Hedyotidis Herba group， so the optimal concentration of Scutellariae Radix-Hedyotidis Herba was set at 40 mg·L-1 for follow-up experiments. As demonstrated by the comparison with the blank group， cisplatin at each concentration inhibited the cell proliferation in a time-dependent manner （P<0.05）. Considering the cell survival rate， the best concentration of cisplatin was set at 10 mg·L-1. Compared with the blank group， Scutellariae Radix-Hedyotidis Herba combined with cisplatin remarkably inhibited the proliferation of A549 cells in a time-dependent manner （P<0.05）， and the differences between the combined medication group and the other two groups became more significant after 72 h of medication （P<0.01）. The IL-6 level in each experimental group， especially in the combined medication group， significantly declined in contrast to that in the blank group （P<0.01）. The mRNA expression levels of HIF-1α and Cyclin D1 in all experimental groups were obviously lower than those in the blank group， with the most significant changes observed in the combined medication group （P<0.05，P<0.01）. The protein expression levels of PI3K， p-Akt， mTOR， HIF-1α， and Cyclin D1 in each experimental group was significantly down-regulated（P<0.05，P<0.01）， and the levels in the combined medication group were even lower than those in the cisplatin group （P<0.01）.ConclusionScutellariae Radix-Hedyotidis Herba has an inhibitory effect on the proliferation of A549 cells， which may be related to its inhibition against the expression and secretion of IL-6/PI3K/Akt/mTOR-HIF-1α axis.
Abstract：ObjectiveTo explore the effect of Duanteng Yimu decoction （DTYM） on the activation of the human umbilical vein endothelial cell （HUVEC） model and the effect on related activated proteins and vascular endothelial growth factor （VEGF） signaling pathway.MethodAfter DTYM （200， 400 g·mL-1） treatment of HUVEC induced by VEGF and tumor necrosis factor-α （TNF-α）， cell proliferation， migration， and tubulogenesis were detected by cell counting kit-8 （CCK-8） assay， 5-ethynyl-2'-deoxyuridine （EdU） assay， transwell migration assay， phalloidin staining， and matrix gel card method. The mRNA expression of adhesion factors， including E-selectin， intercellular adhesion molecule-1 （ICAM-1）， and vascular cell adhesion molecule-1 （VCAM-1） was detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The expression of von Willebrand factor （VWF）， platelet-endothelial cell adhesion molecule-31 （CD31）， angiogenic factor cysteine-rich-61 （CYR61）， angiopoietin-1 （ANG-1）， VEGF， and VEGF receptor-2 （VEGFR2） was detected by Western blot. Immunofluorescence was used to determine CD31 expression.ResultCompared with the normal group， the model group showed potentiated proliferation， migration， and tubulogenesis of HUVEC （P<0.05， P<0.01）， elevated mRNA expression of E-selectin， ICAM-1， and VCAM-1 （P<0.01）， up-regulated protein expression of VWF， CD31， ANG-1， CYR61， VEGF-α， and phospho （p）-VEGFR2 （P<0.05， P<0.01）， and increased CD31 immunofluorescence intensity （P<0.01）. Compared with the model group， the DTYM groups displayed blunted proliferation， migration， and tubulogenesis of HUVEC （P<0.05， P<0.01）， decreased mRNA expression of E-selectin， ICAM-1， and VCAM-1 （P<0.05， P<0.01）， down-regulated protein expression of VWF， CD31， ANG-1， CYR61， VEGF-α， and p-VEGFR2 （P<0.05， P<0.01）， and weakened CD31 immunofluorescence intensity （P<0.01）.ConclusionDTYM inhibits HUVEC proliferation， migration， adhesion， and tubulogenesis， which is associated with the regulation of CD31， VWF， CYR61， and ANG-1 expression in HUVEC and the VEGF signaling pathway.
Abstract：ObjectiveTo investigate the protective effect of Naoxin'an capsule （NC） against glial cell activation and inflammatory damage in brain of rats with chronic cerebral hypoperfusion-induced vascular cognitive impairment （VCI）.MethodOne hundred and fifty rats were randomly divided into a sham operation group （n=20） and a modeling group （n=130）. Following the modeling with the two vessels occlusion （2-VO） technique， 87 successfully modeled rats were randomly divided into the model group， positive drug group （aricept， 0.5 mg·kg-1）， and low-， medium-， and high-dose （0.18， 0.36， 0.72 g·kg-1） NC groups， with 17-18 rats in each group. After intragastric administration of NC for eight weeks， the Morris water maze test and passive avoidance test were conducted to detect the effects of NC on learning and memory ability of VCI rats. Changes in neuronal structure of rat hippocampal CA1 area were observed by hematoxylin-eosin （HE） staining， and the neuronal apoptosis in hippocampus by terminal deoxynucleotidyl transferase （TdT）-mediated dUTP nick end labeling （TUNEL） staining. Western blot assay was used to detect the expression levels of glial fibrillary acidic protein （GFAP）， ionized calcium-binding adapter molecule 1 （Iba-1）， phosphorylated p38 mitogen-activated protein kinase （p38 MAPK）， and phosphorylated nuclear factor κB （NF-κB）， followed by the measurement of interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α） in the brain by enzyme-linked immunosorbent assay （ELISA）.ResultCompared with the sham operation group， the model group displayed obviously decreased spatial learning and memory ability and memory retention ability （P<0.05， P<0.01）， neuronal damage in hippocampal CA1 area， enhanced neuronal apoptosis （P<0.01）， up-regulated GFAP and Iba-1 （P<0.01）， elevated phosphorylation of p38 MAPK and NF-κB （P<0.01）， and increased IL-1β and TNF-α （P<0.01）. Compared with the model group， NC at each dose significantly improved the spatial learning and memory ability and memory retention ability of VCI rats （P<0.05， P<0.01）， ameliorated the neuronal damage in hippocampus CA1 area， reduced the apoptosis rate of nerve cells （P<0.05， P<0.01）， down-regulated the expression of GFAP and Iba-1 （P<0.01）， decreased the phosphorylation levels of p38 MAPK and NF-κB （P<0.05， P<0.01）， and lowered TNF-α and IL-1β levels （P<0.01）.ConclusionNC alleviates the inflammatory damage of the central nervous system caused by activated p38 MAPK and NF-κB and improves chronic cerebral hypoperfusion-induced VCI in rats by inhibiting the activation of microglia and astrocytes.
Abstract：ObjectiveTo explore the effects of Xintongtai （XTT） on traditional Chinese medicine （TCM） syndrome score and collagen fibers in vascular smooth muscle cells（VSMCs） of rabbits with atherosclerosis in the regulation of p38 mitogen-activated protein kinase （p38 MAPK）/activator protien-1 （AP-1）signaling pathway.MethodA total of 120 rabbits of SPF grade were randomly divided into the sham operation group， combined phlegm and blood stasis model group， rosuvastatin group， and low-， middle-， and high-dose XTT groups. The rabbit model of atherosclerosis due to combined phlegm and blood stasis was established by exposing them to high-fat diet and balloon injury. Following modeling， the corresponding drugs were administered by gavage for eight weeks （2.3， 4.6， 9.2 g·kg-1 for low-， middle-， and high-dose XTT groups and 0.55 mg·kg-1 for rosuvastatin group）. At the end of medication， the abdominal aorta was isolated and stained with htoxylin-eosin （HE） for observing the vulnerable plaque. Matrix metalloproteinase-9 （MMP-9） and tissue inhibitor of metalloproteinase-1 （TIMP-1） were detected by immunohistochemistry （IHC）. The collagen fiber decomposition in VSMCs was observed after Masson staining. The protein expression levels of p38 MAPK and AP-1 in aorta was assayed by Western blotting. The combined phlegm and blood stasis syndrome was scored based on TCM syndrome scoring scale.ResultCompared with the model group， XTT at each dose and rosuvastatin significantly decreased MMP-9 content， increased TIMP-1， down-regulated p38 MAPK protein expression， and weakened the nuclear translocation of AP-1 （P<0.01）. Compared with the low-dose XTT group， the middle- and high-dose XTT groups and rosuvastatin group exhibited obviously lowered MMP-9，elevated TIMP-1， down-regulated p38 MAPK protein expression， and diminished AP-1 nuclear translocation （P<0.05，P<0.01）. The TCM syndrome scores of the middle- and high-dose XTT groups and rosuvastatin group were significantly improved as compared with that in the model group （P<0.05，P<0.01）. The comparison with the low-dose XTT group revealed a remarkable improvement in TCM syndrome score of the middle- and high-dose XTT groups and rosuvastatin group （P<0.01）. As demonstrated by Masson staining， the smooth muscle fibers in the model group were arranged in disorder， accompanied by enhanced collagen decomposition， thinned fibrous cap， and increased plaque vulnerability. Compared with the model group， the VSMCs in each XTT group and rosuvastatin group were orderly arranged， manifested as decreased collagen fiber decomposition and increased plaque stability.ConclusionXTT down-regulates the expression of p38 MAPK and MMP-9， increases the level of TIMP-1， reduces the nuclear translocation of AP-1， diminishes the decomposition of collagen fibers in VSMCs， and improves the score of combined phlegm and blood stasis syndrome. XTT alleviates arteriosclerosis due to combined phlegm and blood stasis by regulating p38 MAPK/AP-1 signaling pathway and downstream cytokines and stabilizing vulnerable plaques.
Keywords：heart receiving Qi from spleen;Xintongtai;atherosclerosis;p38 mitogen-activated protein kinase（p38 MAPK）/activator protien-1（AP-1） signaling pathway;traditional Chinese medicine syndrome score
Abstract：ObjectiveTo investigate the intervention effect of modified Shengjiangsan on hypoxia-inducible factor-1α （HIF-1α）/nicotinamide adenine dinucleotide phosphate oxidase 4 （NOX4） signaling pathway in membranous nephropathy （MN） rats and to explore its mechanism to reduce oxidative stress and apoptosis in renal tissues.MethodCationized bovine serum albumin （C-BSA） was injected into the tail vein of rats to replicate the MN model. Rats were randomly divided into a model group， a modified Shengjiangsan group， and a benazepril group after modeling， and administered by gavage once a day accordingly. At the end of the 4th week， the 24-h urine total protein （UTP）， urea nitrogen （BUN）， and serum creatinine （SCr） levels of each group were detected. Enzyme-linked immunosorbent assay（ELISA）was used to detect the levels of superoxide dismutase （SOD）， malondialdehyde （MDA）， and reactive oxygen species （ROS） in renal tissues of rats. In situ end labeling（TUNEL） staining was used to detect the cell apoptosis rate. The mRNA and protein expression levels of HIF-1α and NOX4 were detected by real-time fluorescence-based quantitative polymerase chain reaction（Real-time PCR）and Western blot， respectively. The immunohistochemistry method was used to detect the protein expression levels of B-cell lymphomas -2 （Bcl-2）， B-cell lymphomas xl （Bcl-xl）， Bcl-2 associated X protein （Bax）， Bcl-2 cell death regulator antibody （Bim）.ResultCompared with the normal group， the model group showed increased UTP （P<0.05）， decreased SOD， elevated MDA and ROS （P<0.05）， up-regulated mRNA and protein expression of HIF-1α and NOX4 （P<0.05）， enhanced protein expression of Bax and Bim， declining protein expression of Bcl-xl and Bcl-2 （P<0.05）， and increased cell apoptosis in renal tissues. Compared with the model group， the modified Shengjiangsan group and the benazepril group displayed declining UTP （P<0.05）， up-regulated SOD， decreased MDA and ROS （P<0.05）， down-regulated mRNA and protein expression of HIF-1α and NOX4 （P<0.05）， diminished protein expression of Bax and Bim， elevated protein expression of Bcl-xl and Bcl-2 （P<0.05）， and reduced cell apoptosis in renal tissues （P<0.05）.ConclusionThe protective effect of modified Shengjiangsan on the kidney is presumedly achieved by reducing the oxidative stress and apoptosis in renal tissues of MN rats via inhibiting the HIF-1α/NOX4 signaling pathway.
Abstract：ObjectiveTo investigate the effect of Stemona tuberosa alkaloids on the apoptosis of human hepatoma SMMC-7721 cells and the expression of apoptosis-related proteins including B lymphocytoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， and cleaved cysteinyl aspartate-specific protease-3 （cleaved Caspase-3）.MethodSMMC-7721 cells were routinely cultured， passaged， and treated with various concentrations （50， 75， 112， 167， and 250 mg·L-1） of S. tuberosa alkaloids， while those in the blank control group were only treated with 10% fetal bovine serum. The cell proliferation was determined by tetrazolium bromide （MTT） colorimetry and colony assay and the cell apoptosis by Hoechst 33258 staining. The protein expression levels of Bcl-2， Bax， and cleaved Caspase-3 were detected by Western blot.ResultS. tuberosa alkaloids inhibited the proliferation of SMMC-7721 cells， and the inhibition rate was significantly increased in comparison with that in the blank control group （P<0.01）， with the half maximal inhibitory concentrations （IC50） at 24 h， 48 h， and 72 h being （173.36±8.75）， （112.14±16.50）， and （96.41±2.60）mg·L-1， respectively. The cell colony-inhibitory activity was significantly increased in a dose-dependent manner （P<0.01）. Compared with the blank control group， S. tuberosa alkaloids promoted the apoptosis of SMMC-7721 cells， manifested as increased number of apoptotic cells and elevated apoptotic rate （P<0.01）. The typical morphological changes such as brightly blue-fluorescent condensed nuclei， cytoplasmic shrinking， and karyopyknosis were found under the upright fluorescence microscope. As revealed by comparison with the blank control group， the expression of Bcl-2 was significantly down-regulated （P<0.01）， while the protein expression levels of pro-apoptotic protein Bax and cleaved Caspase-3 in the 75， 112， 167， and 250 mg·L-1 S. tuberosa alkaloids groups were significantly up-regulated （P<0.01）.ConclusionS. tuberosa alkaloids inhibit the proliferation of SMMC-7721 cells and promote their apoptosis possibly by inhibiting Bcl-2 protein expression and promoting Bax and cleaved Caspase-3 protein expression.
Keywords：Stemona tuberosa alkaloids;human hepatoma;SMMC-7721 cells;apoptosis;B lymphocytoma-2 （Bcl-2）;Bcl-2-associated X protein （Bax）;cleaved cysteinyl aspartate-specific protease-3 （cleaved Caspase-3）
Abstract：ObjectiveTo investigate the effects of ligustrazine combined with emodin on angiogenesis of ascites carcinoma Walker-256 cells by observing their inhibition against nuclear factor-κB （NF-κB）， hypoxia-inducible factor-1α （HIF-1α）， and vascular endothelial growth factor-C （VEGF-C） in HIF signaling pathway.MethodFifty SD rats were randomly divided into sham operation group， model group， ligustrazine group， emodin group and ligustrazine combined with emodin group. Following the in situ injection of rat ascites carcinoma Walker-256 cells into the liver of normal rats， they were grouped and administered with ligustrazine （10 mg·kg-1）， emodin （10 mg·kg-1）， and ligustrazine （10 mg·kg-1） plus emodin （10 mg·kg-1） for seven days. Afterwards， the tumor-inoculated liver tissue was sampled from the experimental group and prepared into pathological sections for investigating tumor cell survival and VEGF expression. The in vitro hypoxia and hypoglycemia model （oxygen-glucose deprivation model）， hypoxia model， and hypoglycemia model of Walker-256 cells were constructed respectively. In the ligustrazine group， emodin group， and ligustrazine combined with emodin group， three consecutive concentrations that did not affect the proliferation of Walker-256 cells were selected for investigation. The drugs were administered before modeling， and the model treatment lasted for 4 h. The levels of HIF-1α， VEGF-C， and NF-κB in the cell culture supernatant of each group were tested.ResultAfter the rat liver was inoculated with Walker-256 cells， the total liver mass was significantly increased（P<0.05）， higher than that in the ligustrazine group， the emodin group， or the ligustrazine combined with emodin group（P<0.05）. Histopathological examination showed that the response of VEGF expression in the liver tissue of each administration group was lower than that of the model group. At the cellular level， the levels of HIF-1α， VEGF-C， and NF-κB in oxygen-glucose deprivation model of the ligustrazine group and the ligustrazine combined with emodin group were significantly reduced（P<0.05）， exhibiting a certain dose-dependent response， followed by the reduction in the hypoxia model. The levels of HIF-1α and NF-κB in the oxygen-glucose deprivation model and the hypoglycemia model of the emodin（1×10-2，1×10-3 mol·L-1） group and the ligustrazine combined with emodin（1×10-2，1×10-3 mol·L-1） group were significantly reduced， but there was no significant change in VEGF-C level of the hypoxia model of all the administration groups.ConclusionLigustrazine or emodin alone or their combination inhibits the abnormal increase in the weight of rat liver after inoculation with Walker-256 cells and the expression of VEGF in the liver tissue. Ligustrazine and emodin inhibit the protein expression of NF-κB and HIF-1α， thereby reducing the gene and protein expression of metastasis-related target VEGF-A activated by HIF-1α transcription， restricting tumor cell neovascularization， and inhibiting the invasion and spread of ascites carcinoma cells. Among them， ligustrazine has the most significant effect against hypoxia. Glucose interferes with the effect of ligustrazine. The combination of ligustrazine with emodin is conducive to diminishing the intervention of glucose and stabilizing the inhibition against tumor cells.
Abstract：ObjectiveTo investigate the effect of Jianpi Bufei prescription （JPBFP） on airway inflammation， airway hyperresponsiveness （AHR）， and cyclic adenosine monophosphate （cAMP） signaling pathway activity in ovalbumin （OVA）-sensitized and challenged juvenile asthma rats.MethodSeventy-five male SD rats were randomly divided into a blank group （n=15） and an experimental group （n=60）. The rats in the experimental group were sensitized by aluminum hydroxide gel containing 0.2% OVA and stimulated by aerosol inhalation of normal saline containing 1% OVA to induce an asthma model， followed by assignment into the following groups： a model group （n=15）， a JPBFP group （n=15， 8.37 g·kg-1·d-1）， an aminophylline group （n=15， 40 mg·kg-1·d-1）， and a dexamethasone group （n=15， 0.1 mg·kg-1·d-1）. AHR was detected by the pulmonary function analyzer， changes in inflammatory cells by white blood cell （WBC） count and differential blood count in bronchoalveolar lavage fluid （BALF）， and pathological changes of lung tissues by hematoxylin-eosin （HE）， Masson， and periodic acid-schiff （PAS） staining. The interleukin （IL）-4， IL-5， IL-13， interferon （IFN）-γ， and tumor necrosis factor （TNF）-α levels in serum and the cAMP level in plasma were tested by the enzyme-linked immunosorbent assay （ELISA）. Protein kinase A （PKA） expression in lung tissues was detected by immunohistochemistry. The cAMP-response element-binding protein （CREB） mRNA and protein expression in lung tissues was detected by the real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot.ResultCompared with the blank group， the model group showed increased lung resistance， decreased pulmonary compliance （P<0.05）， elevated WBC count and proportion of eosinophils in BALF （P<0.05）， up-regulated levels of IL-4， IL-5， IL-13， and TNF-α in peripheral blood， declining IFN-γ level （P<0.01）， severe pathological changes of lung tissues， dwindled cAMP， and down-regulated PKA and CREB expression （P<0.01）. Compared with the model group， JPBFP inhibited AHR， reduced WBC count and proportion of eosinophils in BALF and lung resistance （P<0.05）， improved pathological changes of lung tissues， increased pulmonary compliance， and up-regulated cAMP in serum and PKA and CREB expression in lung tissues （P<0.01）.ConclusionJPBFP can improve AHR， inhibit airway inflammation， and alleviate lung injury in asthma rats. Its mechanism may be related to the up-regulation of the activity of the cAMP/PKA/CREB signaling pathway.
Keywords：Jianpi Bufei prescription;asthma;airway hyperresponsiveness（AHR）;cyclic adenosine monophosphate （cAMP）;protein kinase A （PKA）;cAMP-response element-binding protein （CREB）
Abstract：ObjectiveTo explore the correlations of the severity of heart failure related to coronary heart disease arising from Qi deficiency and blood stasis with cardiac function indexes， energy metabolism indexes， coagulation function indexes， and inflammatory factors， in order to provide a scientific basis for further research on the biological foundation of this disease.MethodTwo hundred patients with heart failure related to coronary heart disease of Qi deficiency and blood stasis syndrome were collected and then classified into mild， moderate and severe groups according to their scores of Qi deficiency and blood stasis syndrome. Meanwhile， 40 healthy persons confirmed by physical examination during the same period were included into the control group. Such biological indexes as cardiac function indexes， energy metabolism indexes， coagulation function indexes， and inflammatory factors were determined in patients of each group for comparison. Then the Spearman rank correlation analysis was conducted to figure out the correlations between differential indexes and the severity of Qi deficiency and blood stasis syndrome， followed by the determination of risk factors for the severity of Qi deficiency and blood stasis syndrome by ordered logistic regression analysis.ResultThe cardiac function indexes， energy metabolism indexes， coagulation function indexes， and inflammatory factors in patients with heart failure related to coronary heart disease arising from Qi deficiency and blood stasis varied significantly. There were significant statistical differences in the levels of N-terminal pro-B-type natriuretic peptide （NT-ProBNP）， 6-minute walk test （6MWT）， heart-type fatty acid-binging protein （H-FABP）， prothrombin time （PT）， activated partial thromboplastin time （APTT）， tumor necrosis factor-α （TNF-α）， and nitric oxide （NO） among the mild， moderate， and severe groups （P<0.05）. The severity of Qi deficiency and blood stasis syndrome was positively correlated with NT-ProBNP （r=0.144）， PT （r=0.173）， and APTT （r=0.144）， but negatively with 6MWT （r=-0.287）. The 6MWT［odds ratio（OR）=0.995， 95% confidence interval（CI） 0.991-0.998），P<0.01］ and APTT（OR=1.088，95%CI 1.021-1.157，P<0.01） were independent risk factors affecting the severity of heart failure related to coronary heart disease arising from Qi deficiency and blood stasis.ConclusionThe severity of heart failure related to coronary heart disease of Qi deficiency and blood stasis syndrome is closely related to NT-ProBNP， 6MWT， H-FABP， PT， APTT， TNF-α， and NO. Moreover， 6MWT and APTT can be used as independent risk factors to evaluate the severity of patients with heart failure related to coronary heart disease due to Qi deficiency and blood stasis.
Keywords：coronary heart disease;heart failure;Qi deficiency and blood stasis syndrome;biological indexes;correlation
Abstract：ObjectiveTo observe the protective effect of Huangqi Guizhi Wuwutang combined with Shengmaiyin on the heart function in patients with diabetic cardiomyopathy and explore its anti-myocardial fibrosis and anti-inflammatory effects.MethodThe 96 patients were randomly divided into observation group （48 cases） and control group （48 cases）. Both groups were given comprehensive measures to control blood sugar， blood lipids， blood pressure and heart failure. Patients in control group took Tongmai Jiangtang capsule， 3 granules/time， 3 times/day. Patients in observation group took Huangqi Guizhi Wuwutang combined with modified Shengmaiyin， 1 dose/day. The treatment courses were three months in both groups. Left ventricular ejection fraction （LVEF）， early diastolic peak velocity E peak/late diastolic peak velocity A peak （E/A）， left ventricular end diastolic diameter （LVEDd） and cardiac output per stroke （SV） through echocardiography were recorded before and after therapy. Cardiac troponin-I （cTn I）， troponin T （cTn-T）， creatine kinase isoenzyme -MB （CK-MB）， lactate dehydrogenase （LDH）， transforming growth factor-β1 （TGF-β1） before and after treatment ， matrix metalloproteinase-2 （MMP-2）， insulin-like growth factor-1 （IGF-1）， interleukin-6 （IL-6）， IL-1， tumor necrosis factor-α （TNF-α）， N-terminal pro-B-type natriuretic peptide （NT-proBNP）， soluble ST2 （sST2） and galectin-3 （Gal-3） levels were detected. Symptom of cardiac insufficiency and traditional Chinese medicine （TCM） syndrome score were evaluated before and after treatment.ResultThe LVEF and E/A data in observation group were higher than those in control group （P<0.01）. The levels of cTn-I， cTn-T， LDH and CK-MB in the observation group were lower than those in the control group （P<0.01）. After treatment， the levels of TGF-β1， MMP-2， IGF-1， IL-6， IL-1， TNF-α， NT-proBNP， sST2 and Gal-3 in the observation group decreased and were lower than those in the control group （P<0.01）. The clinical efficacy of the observation group was better than that of the control group （Z=1.974，P<0.05）.ConclusionOn the basis of conventional intervention of western medicine， Huangqi Guizhi Wuwutang combined with modified Shengmaiyin has anti-inflammatory and anti-myocardial fibrosis effects， with inhibitory effect on myocardial remodeling， and can reduce myocardial tissue damage to improve ventricular diastolic function and protect heart function. With such high clinical efficacy， it is worthy of clinical use.
Keywords：diabetic cardiomyopathy;Huangqi Guizhi Wuwutang;Shengmaiyin;myocardial fibrosis;inflammatory factors;heart function
Abstract：ObjectiveTo explore the long-term effect of Zhenzhu Tiaozhi capsule（FTZ） on hemoglobin A1c（HbA1c）in patients with type 2 diabetes mellitus （T2DM） based on real-world data.MethodT2DM patients who were provided with FTZ （FTZ group） and those receiving conventional hypoglycemic drugs （control group） were extracted from the hospital information system （HIS） of the First Affiliated Hospital of Guangdong Pharmaceutical University， followed by propensity score matching （PSM） for balancing the confounding factors between groups. With HbA1c as the efficacy evaluation index， the difference in efficacy between the two groups was compared using t-test and χ2 test. For repeated measurement data of the same patient， the difference in efficacy and the stability of FTZ against HbA1c were analyzed by generalized estimating equation （GEE）. The factors that might affect the efficacy of FTZ against HbA1c were subjected to multivariate linear regression analysis （MLRA）， and the subgroup analyses were then conducted after the stratification of relevant factors.ResultThere were 46 patients included in the FTZ group and 1 208 patients in the control group. PSM yielded 42 pairs of samples with balanced covariates between groups. As revealed by one-year observation， ① HbA1c in the FTZ group after treatment was 6.51%±1.09%. No significant difference was observed either in pre- and post-treatment comparison in the FTZ group or in its comparison with the control group. At the same time， the HbA1c compliance rate in the FTZ group was 73.8% after treatment. No significant difference was observed either in pre- and post-treatment comparison in the FTZ group or in its comparison with the control group. ② The GEE results showed that the post-treatment HbA1c levels in the two groups were not significantly different from each other. Moreover， the HbA1c level remained stable over treatment time. ③ MLRA and subgroup analyses results demonstrated that FTZ was more effective in patients with high baseline HbA1c ［β=-0.530，95% confidence interval（CI） -0.850～-0.209，P<0.01］ or those who were complicated with hypertension （β=-0.918，95%CI -1.614～-0.222，P<0.05）.ConclusionIn the real world， FTZ is able to control the blood sugar， and its effect is similar to those of conventional hypoglycemic drugs. Besides， it is capable of stabilizing the blood sugar for a long time.
Abstract：ObjectiveTo investigate the clinical effect of Tiaomai mixture combined with metoprolol tartrate on premature ventricular contraction in coronary heart disease （CHD） due to Qi-Yin deficiency and stagnated heat in blood vessel.MethodA total of 95 patients with CHD complicated with premature ventricular contraction were randomized into a treatment group and a control group. Four cases dropped out， leaving 91 cases （45 in the treatment group and 46 in the control group） included in the follow-up. On the basis of routine treatments for CHD， patients in the control group were further treated with metoprolol tartrate， while those in the treatment group received metoprolol tartrate plus Tiaomai mixture. Such curative effect and safety indexes as traditional Chinese medicine （TCM） syndrome score， electrocardiogram （ECG）， and 24 h dynamic ECG were observed before and after four-week treatment.ResultAfter treatment， the therapeutic effect on arrhythmia in the treatment group was better than that in the control group（P<0.05）. The treatment group was superior to the control group in reducing the frequency of premature ventricular contraction （P<0.05）， improving the Lown grade （P<0.01）， increasing the heart rate variability index （P<0.05）， and ameliorating the QT dispersion in ECG （P<0.05）， hypersensitive C-reactive protein， and homocysteine（P<0.05）. As revealed by comparison with those before treatment， both interventions improved TCM syndrome， with better outcomes observed in the treatment group （P<0.01）， manifested as the alleviation of shortness of breath， fatigue， dry mouth with desire to drink， and tongue and pulse manifestations （P<0.01）.ConclusionTiaomai mixture improves the clinical efficacy against arrhythmia in CHD patients by regulating the heart rate variability index， inhibiting inflammatory cytokines， lowering homocysteine， and relieving clinical symptoms， which is worthy of clinical promotion and application.
Keywords：Tiaomai mixture;premature ventricular contraction;cooling blood and clearing heat;heart rate variability;hypersensitive C-reactive protein
Abstract：ObjectiveTo observe the regulatory effect of modified Shengmaiyin adjuvant chemotherapy on the nutritional status and immune function of children with acute lymphocyte （ALL）.MethodOne hundred children with ALL chemotherapy were randomly divided into observation group and control group. Both groups were given VDLD regimen as the induction therapy and CAM regimen as the early therapy. Control group was given Shengmaiyin orally， 10 mL/time， 3 times/day. Observation group was given modified Shengmaiyin orally， 1 does/day. The course of treatment for both groups was 3 months. Micro-nutrition assessment （MNA） was carried out before and after treatment， and serum total protein （TB）， albumin （ALB）， prealbumin （PAB）， T cell CD4+， CD8+， immunoglobulin G （IgG）， IgM， IgA levels were detected before and after treatment. The ratio of CD4+/CD8+ was calculated. The traditional Chinese medicine （TCM） syndrome before and after treatment and the piper fatigue scale （PFS-R） and universal core scale of children's quality of life （PedsQL） were scored. The changes of white blood cells （WBC）， red blood cells （RBC）， hemoglobin （Hb） and platelets （PLT） were evaluated before and after treatment.ResultThe nutritional status of the observation group was better than that of the control group after treatment （Z=2.018， P<0.05）. The observed fatigue was lighter than that of the control group （Z=2.029， P<0.05）. The MNA score of the observation group was higher than that of the control group （P<0.01）. The scores of PFS-R and deficiency of both Qi and blood were lower than those of the control group （P<0.01）. The CD4+ levels and CD4+/CD8+ levels of TB， ALB and PAB in the observation group were higher than those in the control group （P<0.01）， while the CD8+ was lower than the control group （P<0.01）. The IgM and IgA levels in the observation group were higher than those in the control group （P<0.01）. The RBC， Hb and PLT levels of the observation group were higher than those of the control group （P<0.01）.ConclusionModified Shengmaiyin can be used as adjuvant therapy for children with ALL chemotherapy by improving nutritional status， immune function and immune balance， reducing clinical symptoms， promoting the recovery of hematopoietic system and improving the quality of life.
Keywords：acute lymphocytic leukemia;deficiency of both Qi and blood;Shengmaiyin;nutritional status;fatigue;immune function
Abstract：ObjectiveBased on pharmacokinetics， the antitussive and expectorant related quality markers （Q-marker） of Trichosanthis Fructus were screened from diosmetin-7-O-glucopyranoside， diosmetin， apigenin， vanillic acid and cinnamic acid， and the candidate Q-marker was evaluated by multivariate statistical method.MethodSix healthy rats were randomly selected and the 70% ethanol extract of Trichosanthis Fructus （dose of 20 g·kg-1） was given by intragastric administration. Blood was collected from the orbital vein at different time points， and the plasma concentrations of 5 components （diosmetin-7-O-glucopyranoside， diosmetin， apigenin， vanillic acid and cinnamic acid） from Trichosanthis Fructus were detected simultaneously by high performance liquid chromatography-triple quadrupole tandem mass spectrometry （HPLC-QqQ-MS/MS）. The main detection conditions were as following：mobile phase of 0.2% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-4 min， 6%-23%B； 4-5 min， 23%-59.5%B； 5-10 min， 59.5%-60%B）， flow rate of 0.5 mL·min-1， the detection wavelength at 254 nm， electrospray ionization （ESI）， positive ion mode detection， multiple reaction monitoring （MRM） mode scanning， scanning range of m/z 50-1 500. Diosmetin-7-O-glucopyranoside， diosmetin， apigenin and vanillic acid with clear pharmacokinetic behaviors were selected as candidate Q-marker about antitussive and expectorant of Trichosanthis Fructus. The contents of these components in 9 batches of medicinal materials were determined and the main detection conditions were the same as the pharmacokinetic study. SPSS 21.0 was used for cluster analysis and principal component analysis （PAC） based on the results of determination.ResultThe pharmacokinetic results showed that the area under concentration-time curve （AUC0-t） of 4 components （diosmetin-7-O-glucopyranoside， diosmetin， apigenin and vanillic acid） were （111.28±9.94）， （27.08±2.76）， （1 376.12±101.86）， （631.32±64.72） μg·h·L-1， respectively. The 9 batches of Trichosanthis Fructus samples were clustered into 3 groups by systematic cluster analysis. The clustering results were related to the variety of Trichosanthis Fructus and also affected by the origin. The PCA results showed that the comprehensive scores of Gaotang Trichosanthis Fructus， Shanxi Trichosanthis Fructus， Hebei Ben Trichosanthis Fructus were 1.919， 1.356 and 0.299， respectively， ranking in the top 3 among all samples. The comprehensive scores of Nongkeyuan No. 1， Hebei Trichosanthis Fructus and Nongkeyuan No. 2 were -0.804， -1.085， -1.120， respectively， which were in the last 3 positions among all samples.ConclusionThe pharmacokinetic characteristics and quality evaluation of diosmetin-7-O-glucopyranoside， diosmetin， apigenin and vanillic acid meet the requirements about antitussive and expectorant related Q-marker of Trichosanthis Fructus.
Keywords：Trichosanthis Fructus;pharmacokinetics;High performance liquid chromatography-triple quadrupole tandem mass spectrometry （HPLC-QqQ-MS/MS）;systematic cluster analysis;principal component analysis （PCA）;quality markers （Q-marker）;index ingredients
Abstract：ObjectiveTo systematically evaluate the safety of heavy metals in Dendrobii Officinalis Caulis and its rhizosphere soil and bedrock in epiphytic culture imitated wild rock fissure. The distribution characteristics of heavy metals in carbonate-black limestone-Dendrobii Officinalis Caulis system in the study area were analyzed.MethodSamples of biennial Dendrobii Officinalis Caulis， black calcareous soil and carbonate rocks were collected from fracture-epiphytic culture in karst area of Guizhou province. The contents of Cu， Pb， As， Cd in Dendrobii Officinalis Caulis， and Cu， Pb， As， Cd， Cr in soil and bedrock were determined by inductively coupled plasma-mass spectrometry （ICP-MS）. The detection conditions were as follows：plasma power of 1 550 W， feedback power of 2 W， sampling depth of 9 mm， atomization chamber temperature at 2 ℃， analysis mode of full quantitative， and double charge of <1.5%. Hg content in Dendrobii Officinalis Caulis was determined by atomic fluorescence spectrometry， and Hg content in soil and bedrock was determined by mercury analyzer. SPSS 26.0 software was used to analyze the test data.ResultThe contents of Cu， Pb， As， Cd and Hg in Dendrobii Officinalis Caulis were all within the safety threshold. The contents of Pb， As， Cd， Hg and Cr in black calcareous soil were higher than the corresponding background values of Chinese soil （P<0.05， P<0.01）， Cd in black calcareous soil was slightly polluted， while Cr， Cu， As， Pb and Hg were clean. The contents of Cu， As， Pb， Cd， Hg and Cr in carbonate rocks were significantly lower than those in black calcareous soil （P<0.01）. The order of heavy metals in black calcareous soil affected by parent rock was Hg>Cd>Cu>As>Cr>Pb. The bioconcentration factor （BCF） of heavy metals in Dendrobii Officinalis Caulis was in the order of Cu>Cd>Pb=Hg>As， but the BCFs of these five heavy metals were all low （all <10%）. The contents of Cu， Pb， Cd and Hg in Dendrobii Officinalis Caulis increased slightly with the increase of heavy metal content in the rhizosphere soil， while the content of As decreased with the increase of As content in the rhizosphere soil. In addition to Cu content in Dendrobii Officinalis Caulis， the migration characteristics of Pb， As， Cd and Hg in the system of carbonate-black limestone-Dendrobii Officinalis Caulis showed consistency.ConclusionThe distribution characteristics of heavy metals in geotechnical plant system in the study area show obvious inheritance. The characteristics of high content， low activity and low pollution risk of heavy metals in black limestone soil and low BCF are the main factors affecting the safety threshold of five heavy metals in Dendrobii Officinalis Caulis.
Abstract：ObjectiveTo compare the chemical constituents of Puerariae Flos from three different varieties of Pueraria montana var. lobata， P. montana var. thomsonii and P. montana var. montana.MethodUltra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MS） was used with the mobile phase of 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （0-20 min， 10%-30%B； 20-30 min， 30%-55%B； 30-35 min， 55%-95%B； 35-37 min， 95%B； 37-40 min， 95%-10%B）， the flow rate was 0.25 mL·min-1. Electrospray ionization （ESI） was used to scan and collect MS data in positive and negative ion modes with scanning range of m/z 50-1 500. The chemical components from different sources of Puerariae Flos were identified in combination with the chemical composition database and literature information. After the obtained data were normalized by MarkerViewTM 1.2.1， they were imported into SICMA-P 14.1 software for principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA） to select the main differentiated components among the three different varieties.ResultA total of 35 compounds were identified from three different varieties of Puerariae Flos， including 22 isoflavones， 6 flavonoids and 7 saponins. The flowers of P. lobata， P. montana var. thomsonii and P. montana var. montana contained 32， 35， 33 compounds， respectively. And 18 differential compounds were screened under the positive and negative ion modes， including kakkalide， tectoridin， 6″-O-xylosyl-tectoridin， 4'-methyltectorigenin-7-glucoside， glycitin， 6″-O-xylosyl-glycitin， irisolidone， kaikasaponin Ⅲ， 6″-O-malonylglycitin， kakkalidone， tectorigenin， rutin， soyasaponin BB， vitexin， biochanin A， genistin， kakkatin， azukisaponin Ⅱ.ConclusionThis research is the first to systematically study the chemical constituents of the flower of P. montana var. montana， although the flower of P. montana var. montana is used as adulterants， it has high contents of tectoridin and 6″-O-xylosyl-tectoridin， which has great potential for development. The efficacy components such as kakkalide and tectoridin in Puerariae Flos from the three sources of varieties are obviously different， and it is necessary to carefully consider the application of these three varieties as Puerariae Flos.
Keywords：Puerariae Flos;Pueraria montana var. lobata;P. montana var. thomsonii;P. montana var. montana;ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MS）;chemical constituents;fragmentation pattern
Abstract：ObjectiveThe correlation between the appearance color of cooked rhubarb samples and the components characterized by high performance liquid chromatography （HPLC） fingerprint was studied to reveal the quality transfer law in the steaming process of processed products with rice-wine.MethodThe visual analyzer was used to analyze the change of the appearance color of cooked rhubarb sample powder at different processing time， the common components and their relative peak areas of processed products with rice-wine were identified by HPLC fingerprint， as well as multivariate statistics and Pearson correlation analysis were used to cluster， discriminate and analyze the appearance color and the component variables in HPLC fingerprint.ResultDuring the processing of cooked rhubarb， the a* （red-green value） of sample powder had no obvious change， but the L* （lightness value）， b* （yellow-blue value） and E*ab （total chromaticity value） showed a decreasing trend， and the appearance color changed from bright to dark， from yellow to brown. A total of 46 chromatographic peaks in the fingerprint were identified at 254 nm and 280 nm， and 18 of them were identified by comparison with reference standards. The change trend of L*， b* and E*ab were positively correlated with the contents of tannins （catechin， epicatechin and ethyl gallate）， stilbene glycoside （trans-3，5，4′-trihydroxystyryl-4′-O-β-D-glucoside）， phenylbutanone glycoside of 4′-hydroxyphenyl-2-butanone-4′-O-β-D-［2ʺ-O-gallic-6ʺ-O-（4ʺ-hydroxy）-cinnamoyl）-glucoside， conjugated anthraquinones （aloe emodin-8-O-glucoside， rhein-8-O-glucoside， emodin-8-O-glucoside） and ω-hydroxyemodin （P<0.05， P<0.01）， and negatively correlated with the contents of free anthraquinones （emodin， aloe emodin and physcion）. Compared with 254 nm， the similarities of chromatographic peaks at 280 nm was more obvious， and the number of detected common peaks was more， which could reflect more subtle differences in chemical composition.ConclusionTannins， stilbene glycosides and phenylbutanone glycosides are strongly correlated with L*， while anthraquinones are strongly correlated with b*， the decrease of E*ab is mainly related to the increase of free anthraquinone content and the decrease of catechins， ω-hydroxyemodin， stilbene glycosides， conjugated anthraquinone and phenylbutanone glycosides. The change of appearance color index of process samples can reflect the change trend of the contents of medicinal components in cooked rhubarb to some extent.
Keywords：Rhei Radix et Rhizoma;steaming with rice-wine;correlation between external appearance color and internal component;high performance liquid chromatography （HPLC） fingerprint;quality control;chromatic value;multivariate statistical analysis
Abstract：ObjectiveTo identify the TIFY gene family in Eucommia ulmoides and analyze its expression from the whole genome level to lay the foundation for further study on EuTIFYs gene function.MethodBased on the E. ulmoides genome database，the TIFY gene family was identified through bioinformatics analysis tools such as National Center for Biotechnology Information（NCBI），MEME，PlantCare，Expert Protein Analysis System（ExPASy），and TBtools. Physicochemical properties，phylogenetic evolution，gene structure，cis-acting elements of the promoters and their expression patterns in leaf development and gum formation of the gene family were systematically analyzed.ResultIn this study，fourteen EuTIFY genes （EuTIFY1-EuTIFY14） were identified in the E. ulmoides genome. The EuTIFYs were composed of 312-1 074 bp nucleotides encoding 102-357 amino acid residues，with isoelectric points of 4.99-10.06 and molecular weight in the range of 10.8-39.14 kDa. According to putative subcellular localization，proteins，which were mainly hydrophilic proteins，localized in the nucleus. The 14 EuTIFYs were unevenly distributed on 13 chromosomes. EuTIFY gene family was divided into four subfamilies： TIFY，JAZ，ZML，and PPD，which contained three，four，five，and two members respectively. The promoters of EuTIFYs contained multiple photoperiodic and abiotic stress-responsive cis-acting elements，which were involved in plant growth and abiotic stress regulation. Expression pattern analysis showed that EuTIFYs exhibited different expression levels in different development stages of E. ulmoides leaves and multiple interactions，and most of the genes were highly expressed in the early stage of leaf development and positively regulated the formation of E. ulmoides gum.ConclusionFourteen EuTIFYs were identified from the whole genome of E. ulmoides，and their structural characteristics and expression patterns were analyzed by bioinformatics. The findings of this study are expected to provide references for further research on the function of EuTIFYs.
Abstract：ObjectiveTo explore the group modules of Chinese medicine and western medicine for the treatment of bronchitis patients with Xiyanping injection based on the real world to provide references for the clinical treatment of bronchitis with Chinese medicine and western medicine.MethodMedical records of 13 874 patients with bronchitis treated by Xiyanping injection were extracted from 29 hospital information systems （HISs） in China，and complex network analysis was carried out using Tabu Search algorithm to obtain the substructure and associated information of core drug combination of Xiyanping injection for the treatment of bronchitis and to analyze clinical medication protocols.ResultMedication protocols for the pathogens of bronchitis are listed below： Xiyanping injection + cefuroxime for bacterial infection. Xiyanping injection + interferon for viral infection. Xiyanping injection + azithromycin for mycoplasma infection. According to the clinical symptoms and complications of bronchitis， the appropriate medication protocols were as follows： Xiyanping injection + ambroxol + Feilike mixture + ibuprofen for uncomplicated bronchitis. Xiyanping injection + ipratropium bromide + budesonide + salbutamol for asthmatic bronchitis. Xiyanping injection + mannitol + furosemide + phenobarbital + gangliosides + immunoglobulin for bronchitis complicated with viral encephalitis. Xiyanping injection + creatine phosphate sodium + vitamin C for bronchitis complicated with viral myocarditis. The combined medication of Chinese medicine for the treatment of bronchitis was adopted based on its characteristics of traditional Chinese medicine.ConclusionThe data of this study were derived from the real world. The combined medications protocols of Xiyanping injection targeting the clinical symptoms and complications of bronchitis were extracted and summarized. However， it is necessary to formulate an individualized medication protocol according to the specific condition.
Abstract：ObjectiveTo explore the effects of Yuanzhisan （YZS） containing Ginseng Radix et Rhizoma （YZSR） or Codonopsis Radix （YZSD） on memory disorder based on network and experimental pharmacology.MethodThe active components and targets of YZS were retrieved from the component database and literature， and the targets of memory disorder from the disease databases. The intersection targets revealed by Veen diagram were subjected to pathway analysis. The common active components of YZSR and YZSD were molecularly docked onto the core targets. Scopolamine hydrobromide was used to establish the memory disorder model， which was employed in the behavioral experiments for evaluating the effect of YZSR and YZSD on memory disorder.ResultThere existed 33 active components for Ginseng Radix et Rhizoma and 31 for Codonopsis Radix， with four common active components and 380 common targets. YZSR contained 85 active components and 790 drug targets， and YZSD 81 active components and 781 drug targets. The mapping of 425 memory disorder targets with those of YZSD and YZSD yielded 133 and 130 intersection targets， respectively. The metabolic pathways involved calcium ion signaling pathway， hypoxia-inducible factor-1 （HIF-1） signaling pathway， mitogen-activated protein kinase （MAPK） signaling pathway， etc. As revealed by molecular docking， the binding energy of common active components to the targets was negative， and the binding effect of frutinone A was the best. Behavioral experiment results showed that both YZSR and YZSD alleviated the memory disorder. In the step-down test， the number of errors in the YZSD group was significant lower than that in the model group （P<0.01）. In Morris water maze test， the movement distance of the YZSD group was remarkably shortened in comparison with that of the model group （P<0.05）. In the open field test， the movement distances of both the YZSR and YZSD group were shortened in contrast to that in the normal group （P<0.05）.ConclusionYZS had a certain effect on memory disorder. There are similarities and differences between YZSR and YZSD in the treatment of memory disorder.
Keywords：Yuanzhisan （YZS）;Ginseng Radix et Rhizoma;Codonopsis Radix;learning and memory;pathway mechanism
Abstract：ObjectiveTo explore the therapeutic principles and medication regularity of Chinese medicine for the treatment of urolithiasis based on data mining and literature research.MethodChinese National Knowledge Infrastructure（CNKI） was searched for the clinical research papers on Chinese medicinal compounds for the treatment of urolithiasis published from database inception to November 10，2020. Therapeutic methods and Chinese medicinal compounds were extracted from the included papers，and the frequencies of therapeutic methods and the drugs in the Chinese medicinal compounds，as well as the efficacies were analyzed. The medication regularity of high-frequency drugs was explored by association rule analysis and cluster analysis.ResultA total of 247 papers and 247 Chinese medicinal compounds were included，and 209 drugs were involved. Those with high frequencies included Lysimachiae Herba，Galli Gigerii Endothelium Corneum，Lygodii Spora，Achyranthis Bidentatae Radix，Pyrrosiae Folium，Plantaginis Semen，and Talcum. The drugs with the highest frequency were effective in promoting urination，draining dampness，and activating blood to resolve stasis，followed by those in tonifying deficiency，regulating Qi，and purgation. Galli Gigerii Endothelium Corneum，Clematidis Radix et Rhizoma，and Succinum were used as specialized drugs targeting the syndrome. Association rule analysis showed that Lysimachiae Herba，Galli Gigerii Endothelium Corneum，and Lygodii Spora were commonly used in pairwise or ternary combination，followed by the pairwise combinations of Lysimachiae Herba with Achyranthis Bidentatae Radix，Pyrrosiae Folium，and Plantaginis Semen. High-frequency drugs were clustered into four categories by clustering analysis. Thirty-one therapeutic principles were employed in the 247 Chinese medicinal compounds. The ones with the highest frequency were clearing heat and eliminating dampness，resolving stasis and promoting urination，and tonifying kidney and replenishing qi.ConclusionFor the treatment of urolithiasis，promoting urination and draining dampness is the core therapeutic principle，and activating blood to resolve stasis is the important therapeutic principle，while tonifying is the potential therapeutic principle. The medication for urolithiasis is featured by specialized drugs targeting the syndrome.
Abstract：Armeniacae Semen Amarum is one of the most commonly used Chinese medicinal materials， with the homology of medicine and food. It is mild toxicity， in addition to raw product， there are blazed and fried Armeniacae Semen Amarum and other processed products. The prescription， process and quality standard of Chinese herbal preparations containing Armeniacae Semen Amarum in the 2020 edition of Chinese Pharmacopoeia （volume Ⅰ）， were studied and analyzed in this paper. The quality control and safety problems of them were discussed， and the proposals were put forward：①To strengthen the quality control of medicinal materials from the origin， including processing， storage， producing area and so on. ②Production technology is the key factor affecting the safety and effectiveness of preparations， so it is necessary to strengthen the control of production process. ③To strengthen the quality research of Armeniacae Semen Amarum preparations， to develop safety quality control projects， and to further improve the quality standards of Armeniacae Semen Amarum preparations. ④To improve the safety tips of Armeniacae Semen Amarum preparations and standardize the instructions. These suggestions can provide reference for scientific understanding and evaluation of traditional Chinese medicine preparations containing Armeniacae Semen Amarum， in order to promote the quality control level of Chinese medicine preparation containing Armeniacae Semen Amarum and its processed products.
Abstract：The small size， moderate drug loading， and targeting properties of nano-preparations make them can be excellent delivery tools for drugs， genes or proteins crossing the cell or blood-brain barrier （BBB）. Currently， facilitating drug crossing BBB with innovative nano-drug delivery systems is considered as a strategic approach for the prevention， diagnosis and treatment of central nervous system （CNS） diseases. However， with the deepening of the research， the adverse reactions and toxicity of nanocarriers have gradually attracted the attention of researchers. Based on this， this paper summarized the situation of BBB-penetrating targeted nano-preparations at home and abroad in recent years from the perspective of classification of types and properties of nanocarriers， and analyzed the advantages and disadvantages of each carrier. The results showed that nano-preparations with active ingredients of traditional Chinese medicine （TCM） as carriers have become a promising way of cancer treatment， but the complexity and diversity of TCM components limited its application to a certain extent. Further studies should be strengthened to lay a foundation for the application and development of TCM nano-preparations in the field of CNS diseases.
Keywords：nano-preparations;blood-brain barrier;targeted drug delivery system;carrier materials;central nervous system （CNS）;liposomes;Chinese materia medica
Abstract：Sepsis， a common critical disease in the intensive care unit（ICU）， features high morbidity and mortality. At present， it is mainly tackled with western medicine， which may trigger a series of problems like antibiotic resistance， adverse hormonal reactions， and high cost after a long-term use. Therefore， exploring new efficient， safe， and cheap drugs and treatment modes has become the focus of our research at this stage. By virtue of unique advantages including "the concept of holism and individualized treatment based on syndrome differentiation"， Chinese medicine has accumulated quite rich experience in the prevention and treatment of sepsis. In recent years， research on the regulation of Chinese medicine on nuclear transcription factor-κB（NF-κB） signaling pathway in sepsis has kept emerging. On this basis， this paper reviewed the etiology and pathogenesis of sepsis， syndrome differentiation and treatment， NF-κB signaling pathway， and its intervention with Chinese medicine. It has been found that some single Chinese herbs and their extracts， Chinese herbal compounds， and Chinese herbal injections effectively inhibit the expression of such inflammatory factors as NF-κB-mediated tumor necrosis factor-α（TNF-α）， interleukin-1（IL-1）， and IL-6 as well as the related proteins， reduce the systemic inflammatory response and organ injury， and improve the prognosis by regulating the activation of NF-κB signaling pathway and the immune function of macrophages. However， due to the limitations of objective conditions， some studies also have the problems of fuzzy pro-inflammatory anti-inflammatory balance mechanism， unclear pharmacokinetics and low drug safety evaluation， which need to be further studied and explored in order to provide a new theoretical basis and diagnosis and treatment thinking for the treatment of sepsis with traditional Chinese medicine.
Keywords：nuclear transcription factor-κB（NF-κB） signaling pathway;traditional Chinese medicine;sepsis;inflammatory factors
Abstract：Psoriasis is an autoimmune disease presented in the context of inflammation， and it mainly results from proliferation and differentiation defects of keratinocytes and abnormal immune response. However， some cellular and molecular mechanisms remain unclear. Although a variety of drugs and physiotherapies are applicable to this disease， they can only be utilized for a short-term period considering their transient effect， high cost， and serious adverse reactions. It is difficult to achieve satisfactory long-term results in the treatment of psoriasis. With the development of network pharmacology and molecular biology and the modernization of traditional Chinese medicine （TCM）， the multi-component and multi-target characteristics of TCM have become prominent， promoting the in-depth research on TCM by doctors and scholars. Nevertheless， there is no detailed summarization on the mechanisms of TCM in interfering with T helper 17 （Th17）/regulatory T （Treg） cell balance to prevent and treat psoriasis. After reviewing the recent literature data， this paper has found that Chinese herbal monomers， active ingredients， and compounds obviously regulate the Th17/Treg axis in psoriasis. Th17 cells have a pro-inflammatory effect， while Treg cells are responsible for maintaining peripheral tolerance. They function in a mutually exclusive manner， and maintaining the Th17/Treg balance helps to effectively reduce inflammatory reaction and regulate immune homeostasis. As revealed by a series of clinical and experimental studies carried out based on the Th17/Treg axis in psoriasis， reducing the percentage of Th17 cells，increasing the percentage of Treg cells，and regulating the levels of related cytokines and transcription factors are conducive to alleviating inflammation and regaining immune homeostasis，which has provided new ideas for further elucidating the pathological mechanism of psoriasis and alternative plans for developing new treatments against psoriasis.
Keywords：traditional Chinese medicine;psoriasis;T helper 17 （Th17）/regulatory T（Treg）;mechanism
Abstract：Glaucoma is a common blinding eye disease characterized by progressive loss of retinal ganglion cells （RGCs） and their axons， gradual loss of visual field， and optic atrophy. The pathological changes of glaucoma are mainly the degeneration， atrophy， and loss of RGCs and their axons， which can eventually lead to the permanent impairment of visual function. The specific pathogenesis of glaucoma remains unclear. Autophagy is a process in which damaged， denatured， or senescent proteins and organelles are transported to lysosomes for digestion and degradation in order to continuously renew and rebuild cells for reuse. As revealed by clinical case analysis and animal experiments， Chinese patent medicine and some traditional Chinese medicine （TCM） therapies may be able to target the autophagy pathway. This paper expounded the role of autophagy in glaucoma-induced ocular hypertension and optic nerve injury from the aspects of stress response of ocular tissue to high intraocular pressure， trabecular meshwork dysfunction， immune regulation， and scar regulation as well as the regulatory effects of some Chinese medicinal ingredients on autophagy， aiming to explore the possibility of integrated TCM and western medicine in regulating autophagy and preventing glaucoma-induced optic nerve injury and early visual field loss. It was found that Chinese medicinal monomers or extracts function via multiple pathways and multiple targets， mainly involving two classical autophagy pathways， namely phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt）/mammalian target of rapamycin （mTOR） and adenosine monophosphate-activated protein kinase （AMPK）. Moreover， the current studies on the neuroprotective effect of TCM mostly focus on the brain and spinal cord lesions of the central nervous system， and its protective mechanism against optic nerve injury in glaucoma still needs further investigation. In addition， autophagy was like a "double-edged sword" in different experimental animal models and methods. How to artificially intervene in autophagy to prevent the apoptosis of RGCs and protect the optic nerve injury is still an urgent problem to be solved in the future research.
Keywords：glaucoma;autophagy;retinal ganglion cells （RGCs）;optic nerve injury;traditional Chinese medicine （TCM） therapies
Abstract：Nardostachyos Radix et Rhizoma is the dry root and rhizome of Nardostachys jatamansi （Valerianaceae） with a long medical history and a broad range of application， which is effective in regulating Qi， relieving pain， resolving depression， and enlivening spleen， as well as dispelling dampness and relieving swelling by external application. It can be used for the treatment of abdominal distension， loss of appetite， and vomiting. Besides， it can also relieve toothache and treat dermatophytosis and pyogenic infection by external use. Moreover， it serves as a common medicinal material in ancient Ayurveda and Unani medical systems in India and also as an ingredient in spices， foods， and cosmetics. Modern pharmacological studies have shown that Nardostachyos Radix et Rhizoma possesses multiple pharmacological activities， such as sedation， anti-epilepsy， anti-convulsion， anti-depression， anti-arrhythmia， anti-malaria， anti-inflammation， anti-bacteria， anti-oxidation， and blood sugar metabolism improvement due to its multiple compounds contained， including terpenes， flavonoids， coumarins， and lignans. The main active components are sesquiterpenoids represented by nardosinone. The pharmacological activities， chemical compositions， and clinical applications of Nardostachyos Radix et Rhizoma have been investigated， but the research on resources， distribution， quality control， cultivation status， and applications are rarely reported. As an important genuine medicinal material from the Qinghai-Tibet plateau， Nardostachyos Radix et Rhizoma is obtained mainly from wild N. jatamansi. Accordingly， the conservation and sustainable utilization of N. jatamansi have attracted much attention all around the world. Based on the resource survey， cultivation research， and relevant literature available， the present study reviewed resources， geographical distribution， chemical compositions， pharmacological activities， quality control， cultivation， and applications of N. jatamansi， aiming to provide references for the conservation and development of N. jatamansi.