Abstract：ObjectiveTo investigate the effect of Danggui Buxuetang（DGBX）on the functional activity of rat endothelial progenitor cells（EPCs）exposed to different luminar shear stress （SS）.MethodEPCs isolated from rat bone marrow were incubated on a parallel plate flow chamber at a steady SS of 0， 0.12， 1.2， 2.4 Pa for 6 h，then the cells exposed to different SS were randomly divided into 8 groups： control group （perfused with serum free medium），simvastatin group（0.1 μmol·L-1 simvastatin），3 DGBX groups（low，medium，high-dose DGBX）and 3 inhibitor groups（3 DGBX groups with LY294002）. After 12 h，the samples were collected for the detection of cell proliferation ，migration，tubule formation ，the secretion of nitric oxide （NO） ，and the expressions of endothelial nitric oxide synthase（eNOS） mRNA and protein kinase B（Akt），respectively.ResultCompared with the control group，simvastatin and DGBX（high-dose）could both promote the functional activities and NO secretion，and up-regulate the expressions of eNOS mRNA and Akt protein in EPCs exposed to different SS（P<0.05），while DGBX（mid-dose）could do these only at 0 Pa. However，LY294002 could inhibit all effects of DGBX on EPCs.ConclusionSS seems to play an important role in the effect of DGBX on EPCs，and DGBX could promote the functional activity of EPCs exposed to SS by up-regulating the expressions of NO/eNOS/Akt.
Abstract：ObjectiveTo study the effects of Fangji Huangqitang（FJHQT） on migration， adhesion，invasion and tube formation of human umbilical vein endothelial cells （HUVECs） induced by vascular endothelial growth factor （VEGF）.MethodHUVECs were induced by VEGF （20 μg·L-1） in vitro. The effects of FJHQT （0.25，0.5，1 g·L-1） on HUVECs were detected by methyl thiazolyl tetrazolium（MTT）， scratch repair， transwell migration， adhesion， invasion and tube formation. Protein in HUVECs was extracted and protein expression levels of phosphorylated Janus kinase 1 （p-JAK1） were detected by Western blot.ResultCompared with control group， VEGF （20 μg·L-1） can increase the proliferation， scratch repair， transwell migration， adhesion， invasion and tube formation of HUVECs cells （P<0.01）， compared with VEGF group， FJHQT （0.25，0.5，1 g·L-1） ，there is no significant effect on the proliferation of HUVECs induced by VEGF for 24 hours， but it can significantly reduce the scratch repair， migration， adhesion， invasion and tube formation of HUVECs induced by VEGF within 24 hours （P<0.05）. Compared with blank group， VEGF could induce abnormal elevation of p-JAK1 in HUVECs （P<0.01）， while FJHQT （0.25，0.5，1 g·L-1） could significantly reduce the expression levels of p-JAK1 （P<0.05，P<0.01）.ConclusionFJHQT can inhibit the migration， adhesion and invasion of HUVECs， the mechanism may be related to JAK1.
Abstract：ObjectiveTo observe the effect of Tongxie Yaofang on the expressions of colon serotonin transporter （SERT）， liver 5-hydroxytryptamine2A receptor （5-HT2AR） protein， serum 5-HT and inflammatory factors in ulcerative colitis （UC） model rats of liver stagnation and spleen deficiency， in order to explore the basis of syndrome of liver stagnation and spleen deficiency and the intervention mechanism of Tongxie Yaofang.MethodFifty male SD rats were randomly divided into blank control group， model group， high， medium and low-dose Tongxie Yaofang group （10，5，2.5 g·kg-1）， and salazosulacil group （0.3 g·kg-1）. The ulcerative colitis model of liver depression and spleen deficiency was established by 2，4，6-trinitrobenzene sulfonic acid （TNBS）/ethanol solution enema + restraint stress + diet loss. After successful modeling， the samples were collected after 21 days of drug intervention. Htoxylin eosin （HE） staining and oil red staining were used to observe the pathological changes of colon and liver in each group. Serum interleukin-6 （IL-6）， IL-9， 5-HT and superoxide dismutase （SOD） were detected by enzyme linked immunosorbent assay （ELISA）. Protein expressions of SERT in the colons and 5-HT2AR in liver of rats were detected by Western blot.ResultCompared with the normal group， obvious ulcers were formed in the colon and lipid droplets in the liver increased in the model group， serum levels of IL-6， IL-9 and 5-HT in the model group increased， while the level of SOD decreased （P<0.05）. The protein expression of SERT in colon decreased， whereas the protein expression of 5-HT2AR in liver increased （P<0.05）. Compare with model group， the pathological damage of colon was improved， and the formation of lipid droplets in liver was reduced in high， medium-dose Tongxie Yaofang groups and sulfasalazine group. The serum levels of IL-6， IL-9 and 5-HT decreased， while the level of SOD increased in Tongxie Yaofang group and sulfasalazine group （P<0.05）. The protein expression of SERT in colon increased in high，low-dose Tongxie Yaofang groups and sulfasalazine group， and the protein expression of 5-HT2AR in liver decreased in medium， low dose Tongxie Yaofang groups and sulfasalazine group （P<0.05）.ConclusionTongxie Yaofang may reduce the content of 5-HT， and regulate the intestinal motility and sensory system by up-regulating the expression of SERT in the colon， inhibit the expressions of IL-6，IL-9 and other inflammatory factors， and play an anti-inflammatory role， reduce the content of 5-HT and the expression of 5-HT2AR in the liver， increase the level of SOD， regulate emotion and lipid metabolism in the liver， and then exert the intervention effect on ulcerative colitis with liver depression and spleen deficiency on the whole.
Abstract：ObjectiveTo explore the mechanism of Suanzaoren Tang in improving learning-memory of sleep-deprived rats based on Nod-like receptor 3 （NLRP3） inflammatome pathway.MethodThe rats were randomly divided into normal control group， model group， Eszolam group（5.4×10-4 g·kg-1·d-1）， low-dose Suanzaoren Tang group（4.59 g·kg-1·d-1）and high-dose Suanzaoren Tang group （18.36 g·kg-1·d-1）. In addition to normal control group， other groups were used to constructed sleep-deprived model， which was concurrent with 30-day continuous drug administration. Water maze was used to evaluate the learning-memory function of rats； The mRNA and protein expressions of NLRP3， apoptosis-related speckle proteins （ASC）， aspartic acid-specific cysteine protease-1 （Caspase-1）， interleukin-1（IL-1） and IL-18 in the hippocampus of rats were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot.ResultCompared with control group， the incubation period of the platform， the total distance of swimming and the duration of first reaching the platform in model group were significantly increased （P<0.01）， while the number of platform crossings and the target quadrant time were decreased （P<0.01）. Compared with the model group， the incubation period， total swimming distance and the duration of first reaching the platform in low-dose Suanzaoren Tang group and high-dose Suanzaoren Tang group were decreased to different degrees （P<0.05，P<0.01）， while the number of platform crossings and the target quadrant time were increased significantly （P<0.05，P<0.01），but with no significant change in estazolam group. Compared with normal control group， mRNA and protein expressions of NLRP3， ASC， Caspase-1， IL-1β， IL-18 in the hippocampus of the model group were significantly increased （P<0.05，P<0.01）. Compared with model group， mRNA and protein expressions of NLRP3， ASC， Caspase-1， IL-1β and IL-18 in the hippocampus of the rats in low-dose Suanzaoren Tang group and high-dose Suanzaoren Tang group were all decreased to different degrees （P<0.05）. The mRNA and protein expressions of NLRP3， ASC， Caspase-1， IL-1β and IL-18 in the hippocampus of Suanzaoren group also decreased， but with no significant change.ConclusionSuanzaoren Tang can improve the learning-memory function of sleep-deprived rats， and its mechanism is related to the inhibition of NLRP3 inflammatome pathway in hippocampus and the alleviation of neuroinflammation.
Abstract：The Shentong Zhuyutang is recorded in an ancient medical book named Yilin Gaicuo， and it is one of the classical prescriptions. Based on the literature reviewed，it is generally found that the Shentong Zhuyutang is evolved from the prescription named Chentongsan that recorded by Danxi Xinfa. Statistics on the dosage of clinical research on this prescription in recent years， reference to the textbook and pharmacopoeia dose， at the same time to respect the original dosage， verify historical changes， and ensure the safety of prescriptions， at the same time consider the pharmacy's dispenses， the author recommends that the clinical dosage of Shentong Zhuyutang could be 3 gram of Gentianae Macrophyllae Radix， 6 gram of Chuanxiong Rhizoma， 10 gram of Persicae Semen， 10 gram of Carthami Flos， 6 gram of Glycyrrhizae Radix Et Rhizoma， 3 gram of Notopterygii Rhizoma Et Radix， 5 gram of Myrrha， 12 gram of Angelicae Sinensis Radix， 5 gram of Cyperi Rhizoma，12 gram of Cyathulae Radix， and 6 gram of Pheretima used as a reference. The efficacy of the compound preparation can be marked as promoting blood circulation， removing blood stasis， removing wind and dampness， and relieving pain. At present， most of its research is focused on clinical research and experimental research， and there are few literature on the research of Shentong Zhuyutang from the level of key information verification. The diseases it treats include more than 10， such as Gubi， Pibi， Xuebi， shechuanchuan， headache， postpartum pain and dysmenorrhea， especially in the study of Gubi， which means that it has a huge market demand and broad development prospects in orthopedic diseases， the core point of "blood stasis and cold dampness evil" should be closely held in the course of treatment. Some scholars deem that Shentong Zhuyutang should not be used the method of Force Sweat in the treatment of Bizheng，while others believe that this prescription should be used with caution in menstruation， pregnancy and blood deficiency. Modern pharmacological studies have shown that Shentong Zhuyutang has obvious anti-inflammatory， relieve pain， anticoagulan， nerve and bone protection. In view of the fact that there are less researches on Shentong Zhuyutang except for Gubi， this suggests that the mechanism of this prescription treat other diseases has a relatively broad research space.
Keywords：classical prescriptions;Shentong Zhuyutang;key information;indications;textual research
Abstract：ObjectiveTo explore the improvement effect of total flavonoids of Mori Cortex combined with total saponins of Anemarrhena Asphodeloide on hyperlipidemia rats with osteoporosis and its possible mechanism.MethodThe 40 SPF male SD rats were adaptively fed for 7 days， and then randomly divided into normal group， model group， calcitriol group （45 ng·kg-1）， total flavonoids of Mori Cortex and total saponins of Anemarrhena Asphodeloide 1∶2 group （0.6 g·kg-1+0.4 g·kg-1） and 2∶1 group （1.2 g·kg-1+0.2 g·kg-1）. Except for the normal group， rats in the other groups were fed with high fat for 9 weeks， the normal group and the model grouotal flavonoids of total flavonoids of Mori Cortex and total saponins of Anemarrhena Asphodeloip were given normal saline by gavage， and the other groups were given corresponding drugs by gavage， after 12 weeks of administration， except for the normal group ， the other groups were given intramuscular injection of glucocorticoids at the same time. After 22 weeks of administration， the weight of rats with total flavonoids from Mori Cortex combined with total saponins of Anemarrhena Asphodeloide was measured. Serum levels of total cholesterol （TC）， triglyceride （TG）， low density lipoprotein cholesterol （LDL-C）， osteocalcin （BGP） and bone alkaline phosphatase （BALP） were determined by biochemical assay. Hematoxylin-eosin （HE） staining to observe the pathological changes of rat tibia. Real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） was used to detect the expression levels of peroxisomal proliferators activate the receptor gamma（PPARγ） and Runt-related transcription factor 2 （Runx2） mRNA in rat bone tissue， immunofluorescence was used to detect the expression of PPARγ and Runx2 in rats.ResultCompared with normal group， the body mass of rats in model group was significantly increased （P<0.01）， and the contents of TC， TG， and LDL-C in the serum were significantly increased （P<0.01）. Compared with model group， the body weight of rats in thet total flavonoids of Mori Cortex and total saponins of Anemarrhena Asphodeloide 1∶2 group and 2∶1 group were significantly reduced （P<0.01）， and the contents of TC， TG， and LDL-C in the serum were significantly reduced （P<0.01）， the content of BGP and BALP increased （P<0.01）. HE staining results showed that compared with the normal group， the tibia fat vacuoles of the model group increased， and the number of osteoblasts decreased， compared with the model group， the total flavonoids of the Mori Cortex and the flavonoids-total saponins of Anemarrhena Asphodeloide 1∶2 group and 2∶1 group decreased in tibia fat vacuoles and increased the number of osteoblasts， the results of immunofluorescence and Real-time PCR showed that， compared with normal group， the expression of Runx2 in the model group decreased and the expression of PPARγ increased （P<0.01）. Compared with model group， the total flavonoids of Mori Cortex-total saponins 1∶2 group and the total flavonoids of Mori Cortex-total saponins 2∶1 Group up-regulated the expression of Runx2 and down-regulated the expression of PPARγ （P<0.05，P<0.01）.ConclusionThe total flavonoids of Mori Cortex combined with the total saponins of Anemarrhena Asphodeloide up-regulated Runx2 and down-regulated the expression of PPARγ mRNA and protein， thereby affecting the metabolism of TG and TC in the blood， achieving a therapeutic effect on osteoporosis， provides experimental basis for the clinical prevention and treatment of hyperlipidemia with osteoporosis.
Keywords：total flavonoids of Mori Cortex;total saponins of Anemarrhena Asphodeloide;hyperlipidemia;osteoporosis
Abstract：ObjectiveTo observe the expression of hepatocyte nuclear factor 1α （HNF1α）， proprotein convertase subtilisin/kexin type 9 （PCSK9） and low-density lipoprotein cholesterol （LDLR） in hypercholesterolemia rat liver， and investigate the mechanism of Shuangyu Tiaozhi Decoction regulating cholesterol metabolism and attenuating hypercholesterolemia.MethodAfter providing a high-fat diet for 4 weeks， 40 SD rats were selected， 8 of which were randomly selected as normal group and fed a normal diet， and the remaining 32 rats were fed a high-fat diet. The rats successfully established as hypercholesterolemic model， were randomized into 4 groups： model group， low dose of Shuangyu Tiaozhi decoction group （7.8 g·kg-1）， high dose of Shuangyu Tiaozhi decoction group （15.6 g·kg-1）， and simvastatin group （4 mg·kg-1）， with 8 rats in each group. The drugs were continuously given for 8 weeks. Serum total cholesterol （TC）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C） were measured. The pathomorphological changes in liver were observed by hematoxylin and eosin （HE） staining. The immunohistochemistry was used to detect PCSK9 and LDLR expression in liver. The mRNA and protein expression levels of HNF1α， PCSK9 and LDLR were determined by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot.ResultCompared with normal group， the TC， TG， LDL-C levels in model group were significantly increased （P<0.01）， the morphology showed obvious liver steatosis. The mRNA and protein expression of HNF1α and PCSK9 were increased （P<0.05）， the mRNA and protein expression of LDLR was decreased （P<0.05）. Compared with model group， the serum TC， TG， LDL-C levels were significantly lowered in the Shuangyu Tiaozhi decoction high-dose group （P<0.01）， the serum TC， LDL-C levels were significantly lowered in the Shuangyu Tiaozhi decoction low-dose group and simvastatin group （P<0.05，P<0.01）， while no significant effect was observed on the serum HDL-C levels in each treatment group. The liver steatosis decreased in each treatment group. The mRNA and protein expression of HNF1α was obviously decreased in each treatment group （P<0.05，P<0.01）， the mRNA and protein expression of PCSK9 was obviously decreased in Shuangyu Tiaozhi decoction low and high-dose groups （P<0.05，P<0.01）， the mRNA expression of PCSK9 was significantly increased in the simvastatin group （P<0.01）， while the protein expression showed a downward trend. The LDLR mRNA levels were significantly increased in each treatment group （P<0.01）， the LDLR protein expression was significantly increased in Shuangyu Tiaozhi high-dose group （P<0.01）， and showed an upward trend in Shuangyu Tiaozhi low-dose group and simvastatin group. Results of immunohistochemistry showed PCSK9 expression was weakly positive， the expression of LDLR was strongly positive in each treatment group. The therapeutic effect of Shuangyu Tiaozhi decoction high-dose group was more remarkable than simvastatin group， while there was no obvious difference between the Shuangyu Tiaozhi decoction low-dose group and simvastatin group.ConclusionShuangyu Tiaozhi decoction may reduce the blood lipid levels through HNF1α/PCSK9/LDLR signaling pathway， play an active role on regulating cholesterol metabolism and alleviating high-fat diet-induced hypercholesterolemia.
Abstract：ObjectiveTo investigate the effect of notoginseng total saponins （TNS） on adriamycin （Adr） resistance in HepG2/Adr cells and the expression and activity of the mechanisms as the modulators of multi-drug resistance， so as to explore the possible mechanism of extracellular signal-regulated kinase （ERK） and protein kinase B （Akt） signaling pathways in reversing the resistance of HepG2/Adr cells mechanism.MethodEffect of TNS on HepG2/Adr cell proliferation was detected by thiazole blue （MTT） method. HepG2/Adr cells were treated with different concentrations （100， 50， 25， 0 mg·L-1） of TNS and （20 μmol·L-1） Adr respectively， and a blank group was set. The high-content screening platform was used to detect the accumulation of Adr in HepG2/Adr cells after 40 minutes， 3 hours and 6 hours. Western blot was used to detect the expression of P-glycoprotein /multidrug resistance/ATP binding cassette subfamily B member 1（P-gp/MDR1/ABCB1） and other drug resistance-related proteins and the main protein expression of ERK/Akt signaling pathway. The change of MDR1 on cell membranes was observed by laser confocal microscopy.ResultCompared with HepG2 cells， the expression of MDR1 in HepG2/Adr cells was significantly increased （P<0.01）. Compared with the Adr group， the half-inhibitory concentration （IC50） of TNS （25， 50， 100 mg·L-1） and Adr （20 μmol·L-1） co-administration group on HepG2/Adr cells in vitro significantly reduced （P<0.01）， and the highest reversal multiple was 10 times. Compared with the Adr group， the co-administration group could significantly increase the accumulation of Adr in the cells （P<0.05） in a dose-dependent manner. Compared with the blank group， the co-administration group could significantly reduce MDR1， ABC semitransporter （ABCG2）， multidrug resistance associated protein （MRP1）， ERK， phosphorylated extracellular regulatory protein kinase （p-ERK）， Akt， phosphorylated protein kinase B （p-Akt）， mammals， rapamycin target protein （mTOR） and phosphorylated mammalian rapamycin target protein （p-mTOR） （P<0.05）， with the same results in the doxorubicin group. Compared with the blank group， there was no significant difference in the distribution and fluorescence intensity of MDR1 on the cell membrane between the Adr group and the notoginseng total saponins （25 mg·L-1） group. Compared with the blank group and the doxorubicin group， TNS could significantly reduce the distribution of MDR1 on the cell membrane （P<0.05）.ConclusionTNS can inhibit the ERK/Akt pathway， reduce the expression of MDR1， and significantly increase the accumulation of doxorubicin in HepG2/Adr cells， which may be one of the mechanisms of notoginseng total saponins in reversing resistance.
Keywords：Notoginseng total saponins;extracellular signal-regulated kinase （ERK）/protein kinase B Akt signaling pathways;reverse drug resistance;P-glycoprotein /multidrug resistance 1/ATP binding cassette subfamily B member 1（P-gp/MDR1/ABCB1）
Abstract：ObjectiveTo investigate the effect of rhein on aquaporin 4 （AQP4） and brain edema after cerebral ischemia and the role of microglia-mediated inflammation in this process.MethodThe modified thread embolization method was selected to establish the cerebral ischemia model of the right middle cerebral artery embolism （MCAO） in rats. The rats were divided into sham operation group， model group， minocycline group， and high， medium and low-dose rhein groups （3.46，1.73，0.865 mg·kg-1）. The neurobehavioral function was measured by a modified neurobehavioral score. Wet and dry weight methods were used to measure the changes of water content in brain tissue of rats with cerebral ischemic injury. Western blot was used to detect the expressions of interferon-γ （IFN-γ） and interleukin-2 （IL-2） in the peripheral ischemic area of rats in each group. Immunofluorescence double labeling method was used to detect the expressions and localization of microglia fine markers Iba-1 and AQP4.ResultCompared with the sham operation group， neurological function score and water content on the side of brain tissue injury of the model group were significantly increased （P<0.05）. Compared with the model group， the neurological function score and the water content of the brain tissue of each drug group were reduced （P<0.05， P<0.01）. Compared with sham operation group， the protein expressions of IFN-γ and IL-2 in the model group increased significantly （P<0.05）. Compared with the model group， the protein expressions of IFN-γ and IL-2 in the peripheral area of cerebral ischemia of each drug group were significantly improved （P<0.05， P<0.01）. Immunofluorescence double staining results showed that compared with the sham operation group， the model group showed significant increase in the fluorescence expression of AQP4 protein on activated microglia， while each drug group could reduce the fluorescence expression of AQP4 protein on activated microglia， different levels of activated microglia markers Iba-1 and AQP4 were co-localized in the peripheral area of cerebral ischemia in each group.ConclusionRhein could reduce the degree of brain edema caused by cerebral ischemic injury， and its mechanism may be related to the inhibition of microglia-mediated neuroinflammation and the down-regulation of AQP4 expression.
Abstract：ObjectiveTo determine the therapeutic effect of in vitro cultivation of bezoar on a mouse model adding disease with syndrome of coronavirus pneumonia with Yidu Xifei syndrome.MethodBALB/c mice were randomly divided into six groups according to their weight grade： normal group， HCoV-229E infection group， cold and damp group， a mouse model combining disease with syndrome of coronavirus pneumonia with Yidu Xifei syndrome， and high and low dose group of in vitro cultivation of bezoar. The combination model of human coronavirus pneumonia with Yidu Xifei syndrome mice was established by the method of cold dampness condition stimulation+coronavirus HCoV-229E infection. In vitro cultivation of bezoar （0.128，0.064 g·kg-1） was administrated by gavage for 3 days from the day of infection. The observation indexes included： general state observation of mice， inhibition rate of lung index and lung index of mice. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the viral load in the lung tissues of mice. Serum levels of motilin（MTL）， gastrin （GAS）， and cytokines interleukin（IL）-10，IL-6， tumor necrosis factor-α（TNF-α）and interferon-γ（IFN-γ） in lung tissue of mice were determined by enzyme-linked immunosorbent assay（ELISA）. The percentages of CD4+ T lymphocytes，CD8+ T lymphocytes and B lymphocytes in the blood of mice were determined by flow cytometry.ResultThe high and low dose group of in vitro cultivation of bezoar can significantly improve the general condition of model mice. Compared with blank group， model group mice lung index increased significantly （P<0.01）， nucleic acids significantly increased expression of lung tissue in mice （P<0.01）， significantly higher serum MTL content in mice， GAS content significantly decreased （P<0.05，P<0.01）， lung tissue cells in the immune factor TNF-α， IL-10 and IL-6 were significantly increased （P<0.01）， peripheral blood lymphocyte CD4+ T cells in mice， The percentages of CD8+ T cells and B cells were significantly decreased （P<0.01）. Compared with model group， in vitro cultivation bezoar mice lung index of high and low dose group were significantly lower （P<0.01）， the lung tissue of mice express nucleic acid decreased significantly （P<0.01）， MTL content decreased significantly （P<0.01）， the lung tissue of mice in the IL-6， IL-10， the TNF-α， IFN-γ levels were significantly lower （P<0.01）， in vitro cultivation bezoar high dose group can significantly increase the CD4+ T cell percentage （P<0.05）， in vitro cultivation bezoar can to a certain extent reduce model mice lung inflammatory exudation， pulmonary interstitial edema， as well as blood stasis symptoms.ConclusionIn vitro cultivation of bezoar has a significant therapeutic effect on a mice model adding disease with syndrome of coronavirus pneumonia with Yidu Xifei syndrome. It can be treated by reducing the lung index of the model mice， improving the pathological damage of the lung tissue， adjusting the immune effective and inhibiting the clearing of inflammatory factors， and to provide a laboratory basis for clinical medication.
Keywords：corona virus disease-2019;in vitro cultivation of bezoar;pathogenesis of traditional Chinese medicine;Yidu Xifei syndrome;immune function;inflammatory factors
Abstract：ObjectiveTo explore the anti-hepatoma effect of compound Phylanthus urinaria Ⅱ （ CPU Ⅱ） by inhibiting the expression of the long non-coding RNA （lncRNA） colon cancer associated transcript-1 （CCAT1） and restoring the expression of microRNA let-7a.MethodReal-time fluorescence quantitative polymerase chain reaction （PCR） was used to detect the expression of lncRNA CCAT1 in normal liver cells （LO2 cells） and hepatocellular carcinoma HepG2 cells， and the differences in expression between these two types of cells were compared. The methylthiazolyl tetrazolium（MTT） assay was used to detect the proliferation of HepG2 cells after treatment with different concentrations of CPU Ⅱ and 5-fluorouracil（5-FU） for 24， 48 and 72 h. Hepatocellular carcinoma HepG2 cells were cultured in vitro and set into three gropes： cell control group， CPU Ⅱ low-dose group （0.8 g·L-1） and high-dose group （1.6 g·L-1）. Real-time PCR was used to detect the mRNA expression of lncRNA CCAT1， microRNA let-7a and its target genes high mobility group protein A2（HMGA2）， and N-RAS in each grope. Western blot was used to detect the protein expression of HMGA2， and Cyclin D1 in each grope.ResultAs compared with LO2 cells， expression of lncRNA CCAT1 in HepG2 cells was significantly up-regulated （P<0.05）. Results of MTT assay showed that the 50% inhibiting concentration（IC50）of CPU Ⅱ and 5-FU on hepatocellular carcinoma HepG2 cells was 1.649， 0.044 648 g·L-1 respectively. As compared with the control group， CPU Ⅱ high-and low-dose groups （1.6， 0.8 g·L-1） significantly inhibited the proliferation of HepG2 cells （P<0.05）， and the effect was most remarkable in CPU Ⅱ high-dose group （P<0.05）. The results of Real-time PCR showed that as compared with control group， the expression of lncRNA CCAT1 mRNA was significantly inhibited in CPU Ⅱ high-and low-dose groups （P<0.05）， and the expression of microRNA let-7a mRNA was obviously up-regulated in high-dose group （P<0.05）， but the expression of HMGA2 mRNA in CPU Ⅱ high-and low-dose groups as well as the expression of N-RAS mRNA in CPU Ⅱ low-dose group were down-regulated （P<0.05）. Western blot results showed that as compared with the cell control group， the protein expression of HMGA2 and Cyclin D1 in CPU Ⅱ high-and low-dose groups （1.6， 0.8 g·L-1） was significantly down-regulated （P<0.05）.ConclusionCPU Ⅱ can inhibit the expression of lncRNA CCAT1， recover the expression of microRNA let-7a， and suppress the mRNA and protein expression of related downstream target genes in hepatoma cells line HepG2， thereby inhibiting the proliferation of hepatocellular carcinoma cells and exerting anti-hepatocellular carcinoma effect.
Keywords：HepG2 cells;Phyllanthus urinaria Ⅱ;colon cancer associated transcript-1 （CCAT1）;microRNA let-7a： high mobility group protein A2（HMGA2）
Abstract：ObjectiveTo observe the clinical efficacy of Sheti Zhiqiu decoction combined with intradermal needling in treatment of allergic rhinitis （AR） of deficient cold of lung Qi syndrome and its effect on cytokines of helper T cell 17 （Th17） and regulatory T cells （Treg）.MethodA total of 105 patients with AR of deficient cold of lung Qi syndrome were randomly divided into traditional Chinese medicine （TCM） group， combination group and western medicine group， with 35 cases in each group. The TCM group was treated with Sheti Zhiqiu decoction， 1 dose a day. The combination group was treated with intradermal needling in addition to the therapy of the TCM group. The selected acupoints were Yintang， bilateral Yingxiang， Fengchi， Feishu and Zusanli. The needles were retained for 3 days before being replaced. The western medicine group was treated with mometasone furoate nasal spray， 100 μg/time at each side of the nasal cavity， qd， and desloratadine citrate tablet， 8.8 mg/time， qd. Three groups were treated for 4 weeks and followed up for 3 months. Nasal and ocular symptom scores and rhinoconjunctivitis quality of life questionnaire （RQLQ） scores of patients in three groups were observed before and after treatment. Levels of serum interleukin-17 （IL-17）， interleukin-10 （IL-10） and transforming growth factor-β1 （TGF-β1） were measured before and after treatment. The clinical efficacy and safety were evaluated， and the disease recurrence rate was observed during the follow-up period.ResultCompared with before treatment， nasal and ocular symptom scores and total score， and RQLQ scores and total score of patients in three groups were significantly decreased after treatment （P<0.01）. Compared with the TCM group and the western medicine group after treatment， except for eyes itching/foreign body sensation/red eyes score， nasal and ocular symptom scores and total score， and RQLQ scores and total score of the combination group were lower（P<0.05，P<0.01）. Compared with before treatment， level of serum IL-17 of patients in three groups was significantly decreased （P<0.01）， while levels of IL-10 and TGF-β1 were significantly increased after treatment （P<0.01）. Compared with the TCM group and the western medicine group after treatment， level of serum IL-17 was lower， whereas levels of IL-10 and TGF-β1 were higher in the combination group （P<0.05，P<0.01）. The clinical efficacy of the combination group was better than that of the TCM medicine group and the western medicine group （Z=-2.207，Z=-2.185，P<0.05）. There was no significant difference in recurrence rate between combination group and the TCM group， and the recurrence rate of both groups was lower than that of the western medicine group（χ2=5.020，χ2=4.835，P<0.05）. There was no significant adverse reaction during the treatment period in three groups.ConclusionSheti Zhiqiu decoction combined with intradermal needling is effective in treatment of patients with AR of deficient cold of lung Qi syndrome. It can significantly relieve patients' symptoms， improve patients' quality of life and reduce disease recurrence. It may play a role by regulating immune balance of Th17/Treg of patients and improving their immune function.
Keywords：allergic rhinitis;deficient cold of lung Qi syndrome;Sheti Zhiqiu decoction;intradermal needling;helper T cell 17;regulatory T cell;cytokine
Abstract：ObjectiveTo study the clinical efficacy of modified Shengyutang on patients with active stage psoriasis vulgaris due to Qi and blood deficiency.MethodThe 134 cases were randomly divided into control group and observation group， with 67 cases in each group. The control group was given avic a capsule + Danggui Buxuewan， while the observation group was given avic a capsule + modified Shengyutang for 4 weeks， respectively. The psoriasis area and severity index （PASI）， dermatological life quality index （DLQI） and psoriasis vulgaris due to Qi and blood deficiency syndrome were observed before and after treatment. The serum growth factor ［endothelial cell specific molecule-1 （ESM-1）， transforming growth factor-β1（TGF-β1）， vascular endothelial cell growth factor （VEGF）］， hemorheological indicators ［high cut blood viscosity （HBV）， low cut blood viscosity （LBV）， erythrocyte sedimentation rate （ESR）］， CC cphenotype receptor（CCR）6， CC cphenotype ligand 20 （CCL20）， monocyte chemotactic protein-4 （MCP-4） in serum and tissue fluid of lesions were detected. Clinical efficacy and recurrence follow-up for 12 months were compared. The safety was evaluated between two groups.ResultThree cases in control group and one case in observation group fell off during the study period. The total effective rate was 96.97% （64/66） in observation group， which was higher than 81.25% （52/64） in control group （χ2=5.064， P<0.05）. During the 12-month follow-up， the recurrence rate was 20.31% （13/64） in observation group， which was lower than 51.92% （27/52） in control group （χ2=6.038， P<0.05）. Compared with control group after treatment， PASI， DLQI， TCM syndromes， ESM-1， TGF-β1， VEGF， HBV， LBV， ESR， CCR6， CCL20 and MCP-4 in observation group were significantly reduced （P<0.05）. No obvious blood and urine routine， or heart， liver and renal dysfunction was observed in the two groups. The incidence of adverse reactions was 3.03% （2/66） in observation group， which was lower than 26.56% （17/64） in control group （χ2=5.764， P<0.05）.ConclusionModified Shengyutang can significantly improve the clinical symptoms of patients with active stage psoriasis vulgaris due to Qi and blood deficiency， with a low recurrence rate and the incidence of adverse reactions.
Keywords：modified Shengyutang;active stage;psoriasis vulgaris;Qi and blood deficiency
Abstract：ObjectiveTo study the clinical efficacy of orthopedics No.1 prescription combined with celecoxib in the treatment of knee osteoarthritis （KOA） with middle stage of cold-dampness syndrome and investigate its effect on serum cytokines levels.MethodThe 72 patients were randomly divided into control group and observation group， with 36 cases each. Patients in both groups were given basic treatment with oral celecoxib capsules （0.2 g/ time， 1 time/day）. On the basis of western medicine treatment， patients in observation group were treated with orthopedics No.1 prescription decoction-free granules by fumigation， 1 bag/time， 1 time/day， 5 times/week. Both groups received treatment for 4 weeks. The visual analog pain score （VAS）， American knee society knee score （KSS）， serum interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， and transforming growth factor-β1 （TGF-β1） levels were observed before and after treatment， and their clinical efficacy was evaluated.ResultAfter treatment， VAS score significantly decreased in both groups （P<0.01）， and KSS score significantly increased （P<0.01）， with better clinical effect in observation group. After treatment， serum IL-1β and TNF-α levels decreased significantly in both groups （P<0.01）， and the levels in observation group were lower than those in control group after treatment （P<0.05）. TGF-β1 content was significantly higher than that before treatment in two groups （P<0.01）.ConclusionOrthopedics No.1 prescription combined with celecoxib for the treatment of KOA with middle stage of cold-dampness syndrome can effectively relieve the clinical symptoms of patients with KOA， improve joint function， improve quality of life， reduce the contents of inflammatory factors IL-1β and TNF-α in serum， and increase the expression of TGF-β1 level.
Abstract：ObjectiveTo observe the effect of modified Shuzhong Huatantang replace hormone on children with rhinosinusitis and adenoid hypertrophy due to spleen deficiency and phlegm obstruction after operation.MethodEighty cases were randomly divided into control group and observation group，40 cases in each group. After nasal endoscopy，the control group was given mometasone furoate，and the observation group was given modified Shuzhong Huatantang for 6 week. The nasal situation Lund-Kennedy assessment scal（Lund-Kennedy），adenoid thickness /nasopharyngeal cavity width （A/N），TCM syndrome were observed for before treatment，6，24，and 48 weeks after operation. The contents of immunoglobulin E（IgE），eosinophilic cationic protein（ECP），eosinophilic granulocyte（EOS），tumor necrosis factor-alpha（TNF-α），interleukin-1β（IL-1β），interleukin-17（IL-17） in serum and nasal secretions were detected before and 48 weeks after operation. The clinical efficacy，complications，recurrence 48 weeks after operation were compared between the two groups.ResultThree cases of abscission in the control group and one case in the observation group during the study period. The total control rate was 94.9% in the observation group higher than that 75.7% in the control group six weeks after the operation（χ2=6.972，P<0.05）. The recurrence rate was 2.6% in the observation group lower than that 18.9% in the control group 48 weeks after operation（χ2=4.137，P<0.05）. To compared with the control group at 6，24 and 48 weeks after operation，Lund-Kennedy，A/N，TCM syndromes in the observation group decrease in the same period （P<0.05）. To compared with the control group 48 weeks after operation，The TNF-α，IL-1β，IL-17，IgE，ECP，EOS in serum and nasal secretions in the observation group were reduced （P<0.05）. The incidence of adverse reactions was 2.7% in the observation group lower than that 29.7% in the control group.ConclusionModified Shuzhong Huatantang can significantly improve the postoperative clinical symptoms with chronic rhinosinusitis and adenoid hypertrophy due to spleen deficiency and phlegm obstruction，and the recurrence rate is lower.
Keywords：modified Shuzhong Huatantang;chronic rhinosinusitis;adenoid hypertrophy;spleen deficiency and phlegm obstruction
Abstract：ObjectiveTo observe clinical effect of addition and subtraction therapy of Zidiantang to purpura nephritis in children （syndrome of blood fever） and regulatory action to immune inflammatory factors.MethodOne hundred and twenty-five patients were randomly divided into control group （61 cases） and observation group （64 cases） by random number table. A total of 57 patients in control group completed the therapy （2 patients were falling off or missing visit and 2 was eliminated）， 59 patients in observation group completed the therapy （3 patients were falling off or missing visit and 2 was eliminated）. Both groups' patients got comprehensive measures of western medicine. Patients in control group got Xueniaoan capsule， 1 to 4 grains/time， 3 times/day. Patients in observation groups got addition and subtraction therapy of Zidiantang， 1 dose/day. The treatment was continued for 2 months and the follow up was recorded for a month. Purpura and urinalysis were recorded for every week. And disappearance of purpura， hematuria and proteinuria were compared for 3 months. Before treatment， and at the first， second and during the follow up， scores of 24 h urine protein quantification （24 hup） and syndrome of blood fever were graded. Levels of microalbuminuria （mAlb）， urinary β2-microglobulin （β2-MG）， cystatin C（CysC）， globulin A1（IgA1）， IgG， complement C3， Th17 cells， Treg cells， interleukin-17 （IL-17）， IL-21， IL-10 and transforming growth factor-β1 （TGF-β1） were detected before and after treatment.ResultAt the first month of treatment， disappearance rate of purpura in observation group was higher than that in control group （P<0.05）. Before treatment， and at the first， second and during the follow up， disappearance rate of hematuria and albuminuria were higher than those in control group （P<0.05）， and scores of 24 h urine protein quantification （24 hup） and syndrome of blood fever were lower than those in control group （P<0.01）. Levels of mAlb， β2-MG， CysC， IgA1， IgG， Th17， Th17/Treg， IL-17， IL-21 and TGF-β1 were lower than those in control group （P<0.01）， and levels of C3， CD4+， Treg and IL-10 were higher than those in control group （P<0.01）.ConclusionOn the basis of conventional western medicine treatment， addition and subtraction therapy of Zidiantang can promote the purpura to subside， improve the syndrome symptoms of blood fever， regulate the immune inflammatory reaction， reduce the inflammatory damage of kidney， thus reduce the hematuria and proteinuria， and play a role in protecting the renal function.
Keywords：purpura nephritis;syndrome of blood fever;Zidiantang;renal function;immunoglobulin;Treg cells;Th17 cells
Abstract：ObjectiveTo discuss clinical effect of Zhuyu Zhixuetang to chronic endometritis （CE） with syndrome of Qi deficiency and blood stasis and to study improvement effect to pregnancy outcome.MethodOne hundred and forty-four patients were randomly divided into two groups， patients in control group were 72 cases and in observation group were 72 cases. In control group， 66 patients completed the therapy because of 4 falling off or missing visit and 2 eliminated， and in observation group， 65 patients completed the therapy because of 3 falling off or missing visit and 5 eliminated. In two groups. anti-infection treatment was gicen to patients. Patients in control group got Fuke Qianjin Pian， 6 tablets/time， 3 times/day. Patients in observation group got Zhuyu Zhixuetang， 1 dose/day. The treatment was continued for 3 months and the follow up was recorded for 6 months. Before and after treatment， changes of menstrual volume， period and cycle were recorded. And hysteroscopy and color Doppler ultrasound of vagina were made， and endometrial morphology and endometrial receptivity were evaluated ［endometrial thickness， resistance index （RI）， pulsation index （PI） and blood flow index （FI）］ were evaluated， and pathology of endometrial were tested. And scores of syndrome of Qi deficiency and blood stasis were graded， levels of interleukin-1 β （IL-1 β）， IL-6 and tumor necrosis factor-α （TNF-α） and T-lymphoid subsets （CD3+， CD4+， CD8+） in peripheral blood were measured. Pregnancy and miscarriage were recorded and the safety was evaluated.ResultAfter treatment， menstrual volume， menstrual period， cycle and complete normal rate of menstruation in observation group were all higher than those in control group （P<0.05）. Endometrial thickness and FI were more than those in control group （P<0.01）， RI and PI were lower than RI and PI in control group （P<0.01）. And compared with control group， levels of IL-1β， IL-6， TNF-α and CD8+ were less （P<0.01）. And levels of CD3+， CD4+ and CD4+/ CD8+ were higher than the data in control group （P<0.01）. During 6 months follow-up， pregnancy rate in observation group was 46.97% （31/66） was higher than 27.69% （18/65） in control group （χ2=5.197， P<0.05）. Total effective rate of endometrial morphology was 96.97% （64/66） higher than 86.15% （56/65） in control group （χ2=4.981， P<0.05）. Total effective rate endometrial pathology was 95.45% （63/66） higher than 84.62% （55/65） in control group （χ2=4.304， P<0.05）. Total effective rate of comprehensive clinical effect was 93.94% （62/66） higher than 81.54% （55/65） in control group （χ2=4.696， P<0.05）. There was no adverse reactions related to traditional Chinese medicine.ConclusionZhuyu Zhixuetang can regulate menstruation， relieve clinical symptoms， improve endometrial morphology under hysteroscopy， regulate systemic and local immune inflammatory response， improve CP， thus improve pregnancy outcome， with better comprehensive effect and safety.
Abstract：ObjectiveTo systematically study the chemical components of Qianyang Yuyin granules and explore its main pharmacodynamic substances and mechanism in the prevention and treatment of hypertensive renal damage.MethodLiquid chromatography coupled with quadrupole time-of-flight mass spectrometry （LC/Q-TOF-MS） was employed to comprehensively analyze the chemical components of Qianyang Yuyin granules. Agilent Poroshell 120 SB-C18 column （3.0 mm×100 mm， 2.7 μm） was used， flow rate was 0.4 mL·min-1， electrospray ionization （ESI） was applied and operated in positive and negative ion modes， the acquisition range was m/z 25-1 000. Mobile phase in positive ion mode consisted of water+10 mmol·L-1 ammonium formate+0.125% formic acid+0.1% methanol （A）-［acetonitrile-water （9∶1）+10 mmol·L-1 ammonium formate+0.125% formic acid］ （B）， and in negative ion mode consisted of water+10 mmol·L-1 ammonium formate+0.1% methanol （A）-［acetonitrile-water （9∶1）+10 mmol·L-1 ammonium formate］ （B） with the gradient elution （0-3.5 min， 5%B； 3.5-4 min， 5%-10%B； 4-9 min， 10%-25%B； 9-18 min， 25%-30%B； 18-25 min， 30%-50%B； 25-27 min， 50%-90%B； 27-32 min， 90%B； 32-33 min， 90%-5%B； 33-39 min， 5%B）. According to the information of the accurate mass， the multistage fragment ions， the mass spectrometric data of the standard substances and the relative reference literature， the structures of the chemical components in Qianyang Yuyin granules were identified. Based on the identified components， network pharmacology study， including target prediction and functional enrichment was applied to screen out the main active substances against hypertensive renal damage， and explore the potential mechanism.ResultA total of 99 chemical components were identified， from which 43 active substances and 48 key targets were screened out. The key components contained kaempferol， quercetin， ferulic acid， luteolin， caffeic acid methyl ester， cinnamic acid， aloe-emodin， emodin， gallic acid， N-trans-feruloyltyramine， isoorientin， 8-O-feruloylharpagide， ethyl caffeate， isookanin， cyasterone， 2，3，5，4'-tetrahydroxystilbene-2-O-β-D-glucopyranoside， loganin， alisol B-23-acetate and harpagide. The key targets included vascular endothelial growth factor A （VEGFA）， serine/threonine protein kinase 1 （AKT1）， Jun proto-oncogene （JUN）， etc.ConclusionQianyang Yuyin granules mainly exert the effects of removing heat from the liver， tonifying the kidney and removing blood stasis via modulation of vascular endothelium， angiogenesis， inflammatory reaction， oxidative stress， immune response and so on.
Keywords：Qianyang Yuyin granules;hypertensive renal damage;network pharmacology;liquid chromatography coupled with quadrupole time-of-flight mass spectrometry （LC/Q-TOF-MS）;pharmacodynamic substances;mechanism;database
Abstract：ObjectiveThis paper aims to clone the cDNA sequence of limonene-3-hydroxylase（StL3OH） in Schizonepeta tenuifolia and analyze its sequence by bioinformatics.MethodSpecific primers were designed based on sequences of StL3OH gene screened from transcriptome sequencing data of S. tenuifolia and the cDNA sequence of StL3OH gene was cloned by reverse transcription polymerase chain reaction （RT-PCR） and analyzed for its bioinformatics.ResultThe StL3OH gene cDNA sequence length was 1 598 bp，containing a 1 497 bp long complete open reading frame which encoded 498 amino acids. StL3OH protein had a theoretical relative molecular mass of 56.40 kDa，with a hydrophilic and unstable nature. Bioinformatics analysis showed that StL3OH protein had no signal peptide but had a transmembrane domain which might be located in endoplasmic reticulum. Multiple sequence alignment and cluster analysis showed that the amino acid sequence of MsL3OH protein had a high similarity with StL3OH protein，both of which contained cytochrome P450 heme binding region，belonging to the D subfamily of cytochrome CYP71 family. Codon bias analysis showed that StL3OH gene preferred guanine/cytosine（G/C） ending codon，with 27 skewed codons， and Nicotiana benthamiana was proven to be the most suitable host for exogenous expression of StL3OH gene.ConclusionThe cDNA sequence of StL3OH gene was cloned from S. tenuifolia for the first time，which will provide a basis for further study on the structure and function of StL3OH protein and the regulation mechanism of StL3OH gene in the accumulation and biosynthesis of monoterpenes in S. tenuifolia.
Abstract：ObjectiveThrough comprehensive evaluation and analysis of the quality of Liuwei Dihuang （LWDH） preparations from different manufacturers and combining factors such as production technology， the key factors in the quality control of LWDH preparations are explored to provide a reference for improving the quality control level of LWDH preparations.MethodMorroniside， loganin and paeonol as quality control markers of LWDH products were determined by high performance liquid chromatography （HPLC）， the mobile phase was acetonitrile （A） -0.3% phosphoric acid aqueous solution （B） for gradient elution （0-5 min， 5%-8%A； 5-20 min， 8%A； 20-35 min， 8%-20%A； 35-45 min， 20%-60%A； 45-55 min， 60%A）， the detection wavelength of paeonol was at 274 nm， and the detection wavelengths of morroniside and loganin were at 240 nm. The quality characteristics of LWDH preparations with different dosage forms （big candied pills， water-honeyed pills， concentrated pills， hard capsules and soft capsules） from different manufacturers were analyzed. Combined these results with their actual production processes， the key-points of quality control in the whole production process were discussed.ResultThe contents of three index ingredients in 128 batches of LWDH preparations were all in conformity with the standards of the 2015 edition of Chinese Pharmacopoeia， however， the content limit of some dosage forms in the current standard was unreasonable. For example， although the daily dose of crude drugs for big candied pills were almost twice the dose of water- honeyed pills （15.00， 8.57 g， respectively）， they got exactly the same daily limits of the contents for both the quality markers. What′s more， these two formulations had the same process， so the differences between the process obviously could not be the reason of these differences.ConclusionIt is recommended that for the products with different dosage forms should have a similar content limits， if there are no obvious distinctions between their production processes. Which may benefit the quality control of the products with multi-dosage forms. The research on the quality standards of proprietary Chinese medicines should deeply study the existing characteristics of the quality standards， and fully respect the laws of the quality attributes of traditional Chinese medicines and the rules of the production process of Chinese patent medicines.
Keywords：Liuwei Dihuang preparations;morroniside;loganin;paeonol;content determination;high performance liquid chromatography （HPLC）;quality control
Abstract：ObjectiveTo investigate the differences of main components of prepared Morindae Officinalis Radix，Morindae Officinalis Radix processed with steaming and salt.MethodA total of 83 batches samples were collected in the market， including 41 batches of prepared Morindae Officinalis Radix，32 batches of Morindae Officinalis Radix processed with steaming and 10 batches of Morindae Officinalis Radix processed with salt. The contents of main components were determined with high performance liquid chromatography coupled with four-pole tandem time-of-flight mass spectrometry（UPLC-Q-TOF-MS），and the differences were analyzed.ResultThe main components of prepared Morindae Officinalis Radix and Morindae Officinalis Radix processed with salt were fructo-oligosaccharides （GFn），monotropein. The main components of Morindae Officinalis Radix processed with steaming were fructose，glucose，sucrose，and monotropein. The main differences of prepared Morindae Officinalis Radix and Morindae Officinalis Radix processed with steaming and salt were the contents of fructose，glucose，sucrose and GF2-GF11. The contents of GF2-GF11 in Morindae Officinalis Radix processed with steaming and salt were all lower than those in prepared Morindae Officinalis Radix，with extremely significant differences（P<0.01）. The contents of fructose，glucose and sucrose in Morindae Officinalis Radix processed with steaming were significantly higher than those in prepared Morindae Officinalis Radix. The content of GF3 in each batch was higher than 40.0 mg·g-1 in prepared Morindae Officinalis Radix and Morindae Officinalis Radix processed with salt，and significantly higher than the limit in Chinese Pharmacopoeia. However，there were only a few batches of Morindae Officinalis Radix processed with steaming in line with the requirements of Chinese Pharmacopoeia. The contents of monotropein in processing Morindae Officinalis Radix and Morindae Officinalis Radix processed with steaming and salt were 42.6，39.8，32.3 mg·g-1，respectively. The content of monotropein in prepared Morindae Officinalis Radix was higher than that in Morindae Officinalis Radix processed with steaming. The content of monotropein in Morindae Officinalis Radix processed with steaming was higher than that in Morindae Officinalis Radix processed with salt. Compared with the components of GF2-GF11，the effect of processing with steaming process and/or salt on monotropein content was relatively less.ConclusionThe contents of GF2-GF11 components in prepared Morindae Officinalis Radix were converted into fructose，glucose and sucrose after processing with steaming and/or salt. The results showed that the content limit of Morindae Officinalis Radix processed with steaming needs to be revised in line with the requirements of Chinese Pharmacopoeia for the quality control of Morindae Officinalis Radix. The results provide a reference basis for revising the quality standards and studying the pharmacodynamic material basis of prepared Morindae Officinalis Radix，Morindae Officinalis Radix processed with steaming and salt.
Keywords：prepared Morindae Officinalis Radix;Morindae Officinalis Radix processed with steaming;Morindae Officinalis Radix processed with salt;component;content;differential analysis
Abstract：ObjectiveTo extract essential oil of Zanthoxyli Pericarpium， to prepare Zanthoxyli Pericarpium essential oil solid preparation and investigate its anti-fungal effect， in order to provide safe， green and efficient fungicide for the storage of Chinese herbal medicine and food.MethodThe essential oil of Zanthoxyli Pericarpium was extracted by steam distillation method， gas chromatography-mass spectrometry （GC-MS） was adopted to analyze the chemical compositions and their relative contents in essential oil of Zanthoxyli Pericarpium from different producing areas， Agilent HP-5 capillary column was used for separation at programmed temperature （the initial temperature was 60 ℃， kept for 2 min， then increased to 280 ℃ by 10 ℃·min-1， kept for 5 min）， the scanning range was m/z 35-590. Zanthoxyli Pericarpium essential oil solid preparation was prepared by nanomolecular sieve adsorption method， and its inhibitory effect on Aspergillus flavus and its conidia was investigated. Ultra-high performance liquid chromatography-fluorescence detector （UPLC-FLD） was used to analyze the inhibitory effect of Zanthoxyli Pericarpium essential oil solid preparation on aflatoxin under the conditions of excitation wavelength of 360 nm and emission wavelength of 440 nm.ResultThe average extraction rate of essential oil in Zanthoxyli Pericarpium from four producing areas was 5.2%. （+）-Limonene， linalool and linalyl acetate were the main components of Zanthoxyli Pericarpium essential oil from different producing areas. When the volume fraction of essential oil in the solid preparation was 0.1%， the inhibition rate of the solid preparation on the conidia of A. flavus was （16.41±8.89）%. When the volume fraction of essential oil in the solid preparation was 0.2%， the inhibition rate for the growth of A. flavus was （8.11±2.70）%. When the volume fraction of essential oil in the solid preparation was 0.5%， the inhibition rate for the growth of A. flavus was （21.62±5.41）%， the inhibition rate for A. flavus conidia was （45.43±5.67）%， and the inhibition effect for the aflatoxin could reach （90.47±12.77）%.ConclusionThere are some differences in the chemical composition of essential oil of Zanthoxyli Pericarpium from different producing areas. Zanthoxyli Pericarpium essential oil has a certain inhibitory effect on the formation of A. flavus conidia and the production of aflatoxin B1. It shows that Zanthoxyli Pericarpium essential oil can be developed into bacteriostatic preparation and used in the storage of Chinese medicinal materials and food.
Abstract：ObjectiveTo analyze active components， its targets and signaling pathways of Shenlian formula based on network pharmacology， and explore the molecular mechanism of Shenlian formula in the treatment of atherosclerotic cardiovascular disease （ASCVD）， in order to provide a basis for the rational interpretation of the prescription compatibility of Shenlian formula.MethodMajor chemical compounds of the formula were obtained by SymMap and Systematic pharmacology database and analysis platform of Traditional Chinese Medicine （TCMSP）， its target proteins were obtained by SymMap and ETCM Databases， and the pathogenic genes responsible for of ASCVD were obtained by DisGeNET and GEO Datebases. Protein targets of drugs and pathogenic genes of diseases were overlapped to obtain predicted targets of Shenlian Formula for ASCVD. Proteins-proteins interactions （PPI） network was built through the String Datebase. The Cytoscape 3.6.0 was used to explore the key compounds and targets of Shenlian formula on ASCVD. Then gene ontology （GO） enrichment and Kyoto Encyclopedia of Genes and Genomes （KEGG） signaling pathway were analyzed to screen out the key targets of Shenlian Formula. Rat I/R model was adopted as representative disease model of ASCVD for experimental verification.ResultThere were 59 candidate compounds， 67 predicted targets and 29 key targets of Shenlian formula on ASCVD. Key targets mainly included cyclooxygenase 2 （PTGS2）， estrogen receptor 1 （ESR1） and TP53. GO analysis showed that the biological functions of potential genes of Shenlian formula in treatment of ASCVD were mainly related to apoptotic， nitric oxide biosynthetic process， response to estradiol， angiogenesis， inflammatory response and oxidative stress and acute-phase response. KEGG pathway enrichment results showed that the pathways of potential genes of Shenlian formula in treatment of ASCVD mainly involved TNF signaling pathway， phosphatidylinositol-3 kinase （PI3K）/ protein kinase B （Akt） signaling pathway， hypoxia induction factor-1 （HIF-1） signaling pathway and apoptosis. Among them， the regulatory effect of Shenlian formula on apoptosis may act on not only TP53， but also different signaling pathways of apoptosis respectively， thus playing a synergistic effect. In vivo experimentation confirmed that Shenlian formula could significantly reduce the myocardial infarction area， improve the myocardial histopathological changes， and especially reduce myocardial mitochondrial injury. Further analysis showed that Shenlian formula can significantly inhibit the expressions of activated proteins in mitochondrial apoptosis pathway.ConclusionAnti-atherosclerosis traditional Chinese medicine Shenlian formula could effectively intervene ASCVD， and its effect on mitochondrial apoptosis of myocardial cells is one of its mechanisms in protecting myocardial ischemia-reperfusion injury.
Keywords：Shenlian formula;atherosclerotic cardiovascular disease （ASCVD）;network pharmacology;mechanism of action
Abstract：ObjectiveTo construct the knowledge base of Tibetan medicine prescriptions and explore to standardize the names of Tibetan medicine prescriptions.MethodBy using the concept of "man-machine combination"，through the construction of Tibetan medicine terminology glossary （data sources： national drug standards，local drug standards，text classics on Tibetan medicines，etc.），the terminology glossary of Tibetan medicine prescriptions was mined. Upon its combination with expert review，the text association between Tibetan medicine prescriptions and various drug standards and dictionaries was constructed，and the standardization methods and techniques of prescription drug names were explored.ResultIn this paper，the Tibetan medicine prescriptions approved for marketing in China were taken as the research object，and various inconveniences caused by the inconsistency between the names of prescriptions and the names of medicinal herbs were revealed. This paper also discussed the design ideas on name standardization of Tibetan medicines from three levels： text association，optimization of evaluation methods，and formation of expert decision-making system. We put forward a five-in-one （screening， evaluation， reviewing， fixing， and renewing） research model of Tibetan medicine name standardization. The construction，functions and advantages of the database and thesaurus of Tibetan medicine prescriptions were described in detail， and in combination with the text notes， association between the standard medicinal materials and the prepared prescriptions was then established.ConclusionThe text association method in this paper can accurately reflect the nonstandard names of Tibetan medicine prescriptions. Combined with expert review，it can be， to a certain extent， extended to the standardization of herb names in prescriptions with large scale of or more complex network structures.
Abstract：ObjectiveTo compare the efficacy and safety of 10 kinds of oral Chinese patent medicines combined with chemotherapy in the treatment of gastric cancer in a network Meta-analysis.MethodPubMed，Embase，Web of Science，Cochrane Library，CNKI，China biomedical literature service system，VIP，Wanfang database were retrieved to obtain Chinese and English literatures published before March 1，2020. The randomized controlled trials（RCTs） and data were screened and extracted. The quality of the included literatures was also evaluated. The pair-work Meta-analysis was conducted by Revman 5.3 software， and the Meta-analysis were conducted by ADDIS 1.16.7 software.ResultA total of 41 randomized controlled trials were eventually involved. All of them were two-arm，including 10 oral Chinese patent medicines，Cinobufotalin，Jinlong capsule，Pingxiao capsule，Shenyi capsule，Xihuang pill （or capsule），Xiaoaiping tablet，Yangzheng Xiaoji capsule，Zhenqi Fuzheng granule combined with chemotherapy for gastric cancer. The effective rate for treating cancer was significantly higher than that of chemotherapy alone （P<0.05）. Zhenqi Fuzheng+chemotherapy was the best choice； the KPS improvement rate of Cinobufacin，Shenyi capsule，Yangzheng Xiaoji capsule，Zhenqi Fuzheng granule combined with chemotherapy in the treatment of gastric cancer was significantly increased（P<0.05）. Shenyi capsule combined with chemotherapy might be the best regimen. The incidence rate of leukocyte decline was significantly lower in the treatment of gastric cancer（P<0.05）. The combination of Zhenqi Fuzheng granule and chemotherapy was the best choice. Traditional Chinese medicine could not reduce the incidence of neurotoxicity（P>0.05）. Cinobufufacin，Yangzheng Xiaoji capsule and Zhenqi Fuzheng granule combined with chemotherapy could increase CD4+/CD8+ in patients with gastric cancer， which was significantly higher than that of chemotherapy alone（P<0.05）.ConclusionAll kinds of oral Chinese patent medicines can play a significantly role in the treatment of gastric cancer. Zhenqi Fuzheng granule，angzheng Xiaoji capsule and Shenyi capsule combined with chemotherapy are more effective in treating gastric cancer.
Keywords：10 kinds of oral Chinese patent medicines;combined chemotherapy;gastric cancer;reticular Meta-analysis;effectiveness;safety
Abstract：In China， about 100 million people currently have chronic obstructive pulmonary disease （COPD）. At the same time， COPD is a multisystem disease， not only affecting the function of musculoskeletal， cardiovascular， kidney and immune systems in patients， but also causing intestinal dysfunction as its extrapulmonary manifestations. From the perspective of traditional Chinese medicine （TCM）， after COPD is formed， deficiency， phlegm stasis and toxicity were accumulated in the lungs， which leads to dysfunction of lung in dispersing and descending， and eventually causes ascending and descending disorder of Qi activities， disorder of fluid supply and distribution， and stagnation of blood stasis. The viscera disease would affect the bowels， and the large intestine is thus affected. Modern medical discovers that， the lungs and intestines have common origins and similar physiological structures， in pathological circumstances， their common mucosal immune system may lead to similar immune factors and inflammatory manifestations in the lungs and intestines. At the same time， the studies have confirmed that there is also a close relationship between intestinal flora and lung， that is "lung-gut axis". These theories partially illustrate the mechanism of COPD in inducing intestinal injury. The specific manifestations of COPD intestinal dysfunction， ① Flora disorder， with increased abundance of intestinal gram-negative bacilli， and inhibited reproduction of Bifidobacterium， Lactobacillus and short-chain fatty acid-producing bacteria. ② Intestinal barrier damage： characterized by the destruction of intestinal epithelium tight connectivity， increased intestinal permeability， and thinning of the mucus layer. ③ Intestinal motility disorder： mostly manifested as weight loss and malnutrition. At present， for the intestinal dysfunction in COPD patients， most of the relevant discussions and targeted treatment methods in TCM are scattered and unsystematic. Guided by the idea of treating different diseases with the same treatment， we summarized the etiology and pathogenesis of COPD intestinal dysfunction by learning from the experience of TCM in treating intestinal flora disorders and inflammatory bowel disease， and proposed preliminary formulation with Tiaoqi Qushi，Tongfu Tongluo as its basic treatment principles in this paper， hoping to provide new ideas for the treatment of COPD.
Keywords：chronic obstructive pulmonary disease;intestinal flora;lung-gut axis;lung-complexed large intestine;intestinal dysfunction;treatment principles and treatment methods;treating different diseases with same treatment
Abstract：Lingnan， locating on the southern coast of China， is a major distribution center of Chinese medicinal materials and imported southern medicine. It is also a well-known patent medicine production base and a foreign trade port. This paper collected relevant historical documents of traditional Chinese medicine （TCM） industry of Lingnan in different times， including Chinese herbs， decoction pieces， Chinese patent medicines and TCM shops. TCM industry of Lingnan originated from the Qin dynasty， and began to develop slowly from the Qin and Han dynasties to the southern and northern dynasties. Private drugstores appeared from the Tang and Song dynasties to the Yuan dynasty， and pharmaceutical commodity production appeared in the Ming dynasty. The TCM industry reached its peak in Qing dynasty， and the modern economic form of the TCM industry appeared during the Republic of China （1912—1949）. It is of great significance for the inheritance， development and innovation of TCM industry of Lingnan to sort out its development history and clarity its development context.
Keywords：Lingnan;traditional Chinese medicine industry;Chinese herbs;decoction pieces;Chinese patent medicines;herbal medicine shop;development history
Abstract：With advanced brewing technology and contemporary table culture， alcohol drinking， which can be traced back to Dukang wine in the Xia dynasty， is very common in China. However， excessive alcohol intake can easily cause alcohol liver damage， ranging from abdominal pain and venous thrombosis to severe hypoglycemia and fat embolism， coma shock and even life-threatening cases. Puerariae Lobatae Radix has a cool property and sweet taste， with functions of antipyretic， promoting the secretion of saliva or body fluid， rash and hangover alleviation， and so on. It was first recorded in Shen Nong's Materia Medica and has been listed as a special anti-alcoholic medicine in traditional Chinese medicine since ancient times. For example， the ancient medical book Compendium of Materia Medica and other records claim that Puerariae Lobatae Radix has the effect of relieving alcohol and protecting the liver. At the same time， Puerariae Lobatae Radix has a long history in both medicine and food. It was listed in the List of Articles That Both Serve as Food and Medicine published by the National Health Commission. Therefore， there are many products containing pueraria for hangover and liver protection. Prior to this， many scholars have carried out relevant researches on the anti-alcoholism efficacy of Puerariae Lobatae Radix， but there is a lack of systematic summaries. The author has consulted relevant domestic and foreign literatures in recent years. The related products were summarized and it was found that the anti-alcoholic effect of pueraria root mainly came from puerarin， pueraria flavonoids and pueraria polysaccharide， puerarin polypeptide， pueraria daidzein and its derivatives， including the main mechanisms such as inhibiting alcohol absorption， accelerating metabolism， anti-oxidation， protection of liver and cardiomyocytes， and neuroprotection. Related products are abundant and well evaluated， but research on related genes needs to be deepened. This article reviews the main anti-alcoholic components， mechanism of action and related products of pueraria， and puts forward suggestions for future research directions， hoping to provide reference for further related research.
Keywords：Puerariae Lobatae Radix;anti-alcoholism;main ingredients;mechanism of action;related products
Abstract：Astragalus membranaceus has the effect of tonifying Qi and solid surface， diuretic support poison， discharging pus and astringent sores to produce muscle. It is not only used for syndromes such as deficiency of lung and temper， deficiency of spleen and diarrhea， but also for stroke， chest obstruction and other diseases. Due to the complex chemical composition and diverse pharmacological effect of Astragalus membranaceus， and the main role in invigorating qi and activating blood circulation has not been clarified. Astragaloside Ⅳ is one of its main active ingredients. In recent years， more and more studies on Astragaloside Ⅳ have been conducted at home and abroad. It has been reported that it has the medicinal value of enhancing immune function， strengthening heart and lowerin blood glucose， diuresis， anti-aging and anti-fatigue， et al， and has extensive pharmacological activity. Among them， the role of cardiovascular and cerebrovascular diseases in particular has attracted increasing attention. Cardiovascular and cerebrovascular diseases are ischemic or hemorrhagic diseases occurring in the heart， brain and systemic tissues due to blood viscosity， atherosclerosis， hypertension， hyperlipidemia， etc.， including cardiovascular and cerebrovascular diseases. Such diseases are a serious threat to mankind and are the leading cause of death worldwide. At present， western medicine is the main treatment， with many adverse reactions and poor long-term prognosis. TCM believes that the imbalance of qi and blood is the basic pathogenesis of this kind of disease. Qi deficiency and blood stasis are more common.At present， Astragaloside Ⅳ in the treatment of cardiovascular and cerebrovascular diseases in a number of studies， and achieved some results， but this review in recent years， Astragaloside Ⅳ in the treatment of cardiovascular and cerebrovascular diseases play the pharmacological activity， in order to explore whether Astragaloside Ⅳ is the main role of astragalus qi to find a theoretical basis for material basis， but also for the innovation of traditional Chinese medicine drug research and development of theoretical basis and practical guidance.
Keywords：astragaloside Ⅳ;cardiovascular;cerebrovascular;invigorating Qi and promoting blood circulation modern pharmacological effect;research progress
Abstract：Zexietang is a Chinese herbal compound prescription with a long history， which consists of Rhizoma alismatis and Atractylodes macrocephala. Zexietang comes from "Synopsis of Golden Chamber"， as "there is a drink under the heart， and its people are bitter and dizzy". Zexietang has the effect of removing water from drinking water and invigorating spleen for diuresis. Modern pharmacological studies have shown that its lipid-lowering and anti-inflammatory effects are very significant. It can be used to treat hyperlipidemia， anti-atherosclerosis and non-alcoholic fatty liver disease. With the continuous development of molecular biology， the research on the pharmacological effects of Zexietang， extracts and their monomers has deepened to the molecular level gradually， and the relevant mechanism of action has also been continuously elucidated. In terms of lipid-lowering effect of Zexietang， the levels of cytokines or receptors such as 3-hydroxy-3-methyl glutaryl coenzyme A reductase （HMGCR）， cholesterol 7α-hydroxylase （CYP7A1）， and liver X receptors （LXR） are affected. It is widely involved in adenosine 5'-monophosphate-activated protein kinase （AMPK） and peroxisome proliferators-activated receptors （PPAR） pathways， while Zexietang's anti-inflammatory effect mainly affects inflammatory factors such as interleukins （IL） and tumor necrosis factors （TNF）， and simultaneously nuclear factor κB （NF-κB）， toll-like receptors （TLR） and other cytokines or receptor-related pathways. In order to promote the further research and clinical application of Zexietang and contribute to the development of modernization of traditional Chinese medicine， the studies of the past 15 years on molecular mechanism of the lipid-lowering and anti-inflammatory effect of Zexietang， Alisma and Atractylodes extract as well as their monomer components were reviewed.
Abstract：Danggui Liuhuang Tang， as one of classical traditional Chinese medicine prescriptions， has been used by major medical experts in clinic since Jin and Yuan dynasties. After review and summarization of relevant literatures on the pharmacological effects and clinical application of Danggui Liuhuang Tang， it was found that Danggui Liuhuang Tang has a wide range of pharmacological activities， and exerts its anti-inflammatory and anti-insulin resistance effects mainly by inhibiting the production of inflammatory factors， such as tumor necrosis factor and interleukin and activating related pathways. In addition， Danggui Liuhuang Tang inhibits the occurrence and development of hepatic fibrosis by attenuating proinflammatory signaling and extracellular matrix accumulation with multiple components， multiple targets， and multiple pathways. Danggui Liuhuang Tang has been widely used in sweat syndromes， with an obvious effect in the treatment of thyroid diseases， diabetes， respiratory tract and other diseases. This paper reviews and summarizes the pharmacological effects and clinical application of Danggui Liuhuang Tang，in an attempt to provide some valuable clues for the subsequent development of Danggui Liuhuang Tang.
Abstract：Sojae Semen Germinatum was firstly recorded in Shennong Bencaojing， and it has a long history of edible and medicinal use. Most ancient medical practitioners described that Sojae Semen Germinatum was processed with black soya bean， while some others recorded that Sojae Semen Germinatum was processed with black soya bean and soybean or with soybean only. In modern times， black soya bean and soybean are both used. Before the Northern and Southern dynasties， the germination process of Sojae Semen Germinatum was mostly soil culture， and then changed into water culture later. The medicinal part of Sojae Semen Germinatum may also change from the initial aboveground part to the whole processed products including the soybean and the bud. The bud length was used to control the processing ending of Sojae Semen Germinatum， but there were different views of the bud length in ancient and modern times. Before the Tang dynasty， Sojae Semen Germinatum was mostly used directly. Since the Tang dynasty， various subsequent processed products of Sojae Semen Germinatum appeared. Most ancient medical practitioners confirmed that Sojae Semen Germinatum was sweet flavor， neutral in nature and non-toxic， and the mainstream believed that it belonged to the spleen， lung and stomach meridians. However， there were different opinions on its efficacy of relieving exterior syndrome by diaphoresis. In this paper， the evolution of Sojae Semen Germinatum was explored after consulting all kinds of ancient books， its historical name， processing history， quality evaluation and others were systematically summarized in order to clarify its historical development and lay a good foundation for the clinical use and further development of Sojae Semen Germinatum.
Keywords：Sojae Semen Germinatum;herbal textual research;name of medicinal material;processing technology;nature， taste and function;quality evaluation;bud length