Abstract：ObjectiveTo explore the effect of modified Xiongxiesan on the proliferation of airway smooth muscle tissues and the expressions of matrix metalloproteinase-9（MMP-9）， tissue inhibitor of metalloproteinase-1 （TIMP-1） in cough variant asthma （CVA） model rats.MethodA total 48 male SD rats were randomly divided into the normal control group （8 rats） and model group （40 rats）. CVA model of rats were established through the intraperitoneal administration with 2 mg ovalbumin （OVA） and 100 mg Al（OH）3， and then aerosol inhalation of 1％ OVA 15 days later. The same volume of sterile saline was given to the normal group through the intraperitoneal injection. Then 40 rats in the modeling group were randomly divided into model group， modified Xiongxiesan group （TCM group， 6 g·kg-1·d-1）， montelukast group （0.4 mg·kg-1·d-1）， chemokine receptor1/2 （CXCR1/2） inhibitor group （G31P group injected subcutaneously via the neck with a dose of 0.5 mg·kg-1 every other day）， and CXCR1/2 inhibitor and modified Xiongxiesan group （G31P+TCM group）， with 8 rats in each group. The control group and the model group were orally given distilled water 10 mL·kg-1·d-1. Then the rats were sacrificed， and lung samples were collected. Histological changes were examined by hematoxylin-eosin（HE）. Basement membrane perimeter （PBM），wall area of bronchial tube （WAt），wall area of bronchial smooth muscle （WAm） and the number of smooth muscle cells （N） were measured using image pro-plus software and standardized based on PBM. The expressions of PCNA， MMP9 and TIMP1 were detected by immunohistochemistry.ResultCompared with the control group， there were a large number of inflammatory cells infiltration and moderate hyperplasia of smooth muscle area in the model group， which however were alleviated in other groups. The expressions of PCNA and MMP-9，TIMP1 were higher in the model group，which were reduced in other groups significantly.ConclusionModified Xiongxiesan can reduce the thickness of airway smooth muscle tissue in the CVA model rats， which may be correlated with the inhibition of the CXCR1/2 pathway， thereby reducing the proliferative activity of smooth muscle tissue and inhibiting the expression of related matrix metalloproteinases.
Abstract：ObjectiveTo study the effect of Yupingfeng San （YPFS） on the expressions of GATA binding protein 3 （GATA3） and forkhead transcription factor3 （Foxp3） in ovalbumin （OVA）-induced lung tissue of allergic rhinitis （AR） mice， and explore the mechanism of YPFS on AR.MethodThe allergic rhinitismice model was established by intraperitoneally injecting with OVA and Al （OH）3， and challenged with OVA intranasally. The mice were divided into four groups： normal，model，chloretadine（3 mg·kg-1） and YPFS（6.5 g·kg-1） group， the corresponding drugs were orally administrated for three weeks. At the end of administration，the infiltration of inflammatory cells， such as mast cells， eosinophils and neutrophils in nasal mucosa， were observed by htoxylin eosin （HE） staining. The serum concentrations of OVA-specific IgE and cytokines ［interleukin-4（IL-4）， IL-5 and γ-interferon （INF-γ）］ were determined by enzyme-linked immune sorbent assay （ELISA）. The expressions of GATA3 and Foxp3 proteins in nasal mucosa tissue were detected by Western blot.ResultThe AR mice had such symptoms as scratching， sneezing and running nose. Nasal mucosa section by HE staining showed significant desquamation of AR mouse nasal mucosa cilia， obvious tissue stromal edema， telangiectasia， and a large number of eosinophilic cells， lymphocytes， plasma cells infiltration. YPFS obviously improved nasal symptoms of allergic rhinitis mice. Nasal mucosa epithelial structure was complete and arranged evenly， with no obvious tissue clearance edema and vasodilation， and inflammatory cell infiltration was significantly reduced. Compared with normal group， the levels of OVA specific IgE， IL-4 and IL-5 in peripheral blood of AR model group were significantly higher（P<0.01）， and the INF-γ level was significantly lower （P<0.01）. Compared with AR model group， the administration of chloretadine and YPFS can significantly reduce the level of OVA specific IgE and IL-4， IL-5， and increase the level of INF-γ in AR mice peripheral blood （P<0.05， P<0.01）. Western blot results showed that compared with normal group， GATA3 protein expression was significantly increased， while Foxp3 protein expression was significantly decreased in AR model group （P<0.05， P<0.01）. Compared with AR model group， YPFS inhibited GATA3， and promoted Foxp3 protein expression （P<0.01）.ConclusionYPFS has an effect in alleviating OVA-induced allergic rhinitis．YPFS may modulate the immune response by regulating the balance of Th2/Treg cells.
Keywords：allergic rhinitis;Yupingfeng San （YPFS）;insulin resistance;immune regulation
Abstract：After a comprehensive analysis and research of ancient books and literatures， we discovered that six Banxia Baizhu Tianma Tang prescriptions have the same name. It was originally created by LI Gao. The idea of composition has a certain inheritance relationship with ZHANG Yuan-su′s Tianma Banxia Tang and Huatan Yuhu Wan， which are recorded in Prescription of Peaceful Benevolent Dispensary. In later generations， four doctors developed five prescriptions of the same name. CHENG Guo-peng of the Qing dynasty reformed the prescription based on the theory of phlegm and dampness. He reduced tonic components based on LI's ideas to strengthen the expectorant function， and then transformed the prescription into two of the same name， one for headache and the other for vertigo. The latter was recorded in the textbook of Chinese Herbal Medicine and The Catalogue of Classic Prescriptions （batch 1）， which has the greatest impact on the contemporary prescriptions. Its composition is Pinelliae Rhizoma Praeparatum Cum Zingbere et Alumine （5.595 g）， Atractylodis Macrocephalae Rhizoma （11.19 g）， Gastrodiae Rhizoma， Poria， Citri Exocarpium Rubrum （each 3.73 g）， Glycyrrhizae Radix et Rhizoma （2 g）， 2 pieces of fresh Zingberis Rhizoma Recens （about 2 g） and 2 Jujubae Fructus. The botanical origin of these herbs is clear， and the dosage of herbs is safe in accordance with the Pharmacopoeia of the People's Republic of China.P. The total amount of each dose is about 37.975 g， while the daily dosage is not far different from that of invention. Since its invention， it has been mainly used in the treatment of phlegm headache and vertigo caused by wind phlegm disturbance. In modern clinical practice， it is widely used in treating various diseases with vertigo and headache as the main symptoms， mainly including hypertension， acute ischemic stroke， vertebrobasilar insufficiency， cervical spondylosis， primary headache， Meniere's disease and benign paroxysmal positional vertigo. When the syndrome differentiation is consistent with relevant syndrome type with phlegm as the main pathological factor in traditional Chinese medicine， we can modify the prescription as appropriate.
Keywords：catalogue of classic prescriptions;Banxia Baizhu Tianma Tang;phlegm damp;vertigo
Abstract：ObjectiveTo study the protective effect and mechanism of artemisinin on systemic inflammatory response syndrome （SIRS）mice using endotoxin （LPS）-induced SIRS mouse model.MethodMale BALB/c mice aged 5-7 weeks were randomly divided into normal group， LPS model group， low， medium and high-dose artemisinin groups （25， 50， 100 mg·kg-1） and ibuprofen group （39 mg·kg-1）. LPS （10 mg·kg-1） was intraperitoneally injected at the 7th day after the prophylaxis. According to the SIRS clinical diagnostic criteria， the respiratory rate， rectal temperature， lung index， spleen index， glycolipid metabolism， brain tissue inflammatory factors， and phosphorylation of lung tissue inflammation-related proteins were measured.ResultIntraperitoneal injection of LPS significantly reduced the respiratory rate of mice （P<0.05）， body temperature decreased significantly （P<0.01）， spleen index increased significantly （P<0.01）， peripheral blood neutrophil percentage increased significantly （P<0.05）， percentage of monocytes decreased significantly （P<0.01）， thrombocyte decreased （P<0.01）， platelet specific ratio decreased （P<0.01）， total cholesterol content in plasma decreased （P<0.01）， plasma glucose content decreased （P<0.01）. The expression of interleukin-1β increased in hippocampus and cortex of brain tissue （P<0.01）， and the secretion of tumor necrosis factor-α increased in hippocampus and cortex of brain tissue （P<0.01）. The expression of phosphorylated protein STAT1 was increased （P<0.01）， and the expression of phosphorylated protein c-Jun was increased （P<0.01）. After the administration of artemisinin， the body temperature and the respiratory rate of mice induced by LPS were significantly increased， the pathological changes of various organs induced by LPS were alleviated， the hypoglycemia induced by LPS was significantly increased （P<0.05）， the levels of inflammatory factors in hippocampus and cortex was significantly reduced， and the expressions of phosphorylated proteins STAT1 and c-Jun in lung tissue were significantly reduced （P<0.05， P<0.01）.ConclusionArtemisinin has a significantly protective effect on SIRS mice induced by intraperitoneal injection of LPS possibly by reducing the secretion of inflammatory factors TNF-α and IL-1β.
Abstract：ObjectiveTo investigate whether ultrafine powder of Gastrodiae Rhizoma （UPG） can alleviate the learning and memory impairment of vascular dementia rats and delay the process of VD， and whether this effect is related to the release of acetylcholine （Ach） through the regulation with acetylcholinesterase （AChE） and choline acetyltransferase （ChAT） and control of cholinergic system.MethodSD rats were randomly divided into sham operation group， UPG low dose group （0.45 g·kg-1）， UPG high dose group （1.8 g·kg-1） and Huperzine A group （80 μg·kg-1）， with 12 rats in each group. The drug administration groups were given orally drugs once a day for 8 weeks， and sham group and model group were given orally the same amount of distilled water. The learning and memory ability of the rats with VD were evaluated by the Morris water maze. Htoxylin eosin（HE） staining was used for pathomorphological observation of hippocampus CA1 area of the rats. The content of Ach was determined by enzyme-linked immunosorbent assay（ELISA）， AChE and ChAT protein expressions were detected by Western blot， and expression of ChAT in hippocampus CA1 area was observed by immunohistochemistry.ResultCompared with the sham operation group， the escape latency of the model group was significantly increased （P<0.01）， and the frequency of crossings platform and the time of staying in the target quadrant were reduced significantly （P<0.01）. HE staining of hippocampal tissues from VD rat showed neuron disorders， loss and degeneration and necrosis， pyknosis of the nucleus and light coloration of the cytoplasm. The level of acetylcholine in the hippocampus was significantly decreased by ELISA （P<0.05）， the expression level of AChE protein was significantly up-regulated， and the expression level of ChAT protein was significantly down-regulated （P<0.01）. Compared with model group， each administration group could significantly reduce the escape latency of the model rats， and significantly increase the frequency of crossing platform and the time of staying in the target quadrant （P<0.01）， the content of Ach was significantly increased （P<0.05）， the expression of AChE protein was significantly down-regulated （P<0.01）， while the expression of ChAT protein was significantly up-regulated （P<0.01）.ConclusionUPG improves the learning and memory ability of vascular dementia rats， and its mechanism may be related to the increase of Ach， ChAT level and the decrease of AChE level.
Keywords：ultrafine powder of Gastrodiae Rhizoma;vascular dementia;learning and memory impairment;acetylcholinesterase （AChE）;choline acetyltransferase （ChAT）
Abstract：ObjectiveTo investigate the effect of ginkgolide B （GB） on the activation of c-Jun aminoterminal kinase（JNK） signaling pathway and apoptosis in amyotrophic lateral sclerosis cell model.MethodNSC34 cells were infected by slow virus containing expression superoxide dismutase1（SOD1）WT and hSOD1G93A and empty plasmid， and screened with a certain concentration of puromycin， so as to observe the transfection efficiency of slow virus and cell morphology under inverted fluorescence microscope. Western blot method was used to verify whether infected cells were over-expressing SOD1 target proteins. The hSOD1G93A-NSC34 cell lines were established and given GB. Cell cultures were divided into normal group， model group and different concentrations of ginkgolide B groups （25， 50， 75， 100 mg∙L-1）. After 48 h， methyl thiazolyl tetrazolium （MTT） was used to detect cell survival rates， and select the best drug concentration. Subsequent experimental groups were divided into normal group， model group， 75 mg∙L-1 GB group， SP600125 group， and 75 mg∙L-1 GB + SP600125 group. Flow cytometry was used to detect the apoptosis of each group of cells. Western blot was used to detect the expressions of phosphorylation（p）-JNK， c-Jun， p-c-Jun， and cysteine aspartic acid protease -3（Caspase-3） proteins.ResultCompared with normal NSC34 cells， hSOD1G93A-NSC34 cell body became round， the synapses decreased and shortened， but the cell morphology of hSODWT-NSC34 cell and empty plasmid group did not change significantly. Western blot showed that hSOD1G93A-NSC34， hSOD1WT-NSC3 intracellular SOD1 protein levels increased significantly （P<0.01）， and the amyotrophic lateral sclerosis cell model was established. Compared with the normal group， the cell activity in the model group was significantly reduced （P<0.01）. Compared with the model group， the cell activity increased at different concentrations of GB， especially when the drug concentration was 75 mg∙L-1 （P<0.01）. In subsequent experiments， compared with the normal group， the apoptosis， and expressions of p-JNK， p-c-Jun， and cleaved Caspase-3 proteins in the model group increased significantly （P<0.01）. Compared with the model group， the apoptosis and p-JNK， p-c-Jun， released Caspase-3 protein expressions of 75 mg∙L-1 GB group， SP600125 group， 75 mg∙L-1 GB + SP600125 group decreased significantly （P<0.05， P<0.01）.ConclusionGB has a protective effect on the cell model of atrophy lateral sclerosis， which may be realized by JNK signal pathway.
Keywords：hSOD1G93A;amyotrophic lateral sclerosis;ginkgolide B;apoptosis;c-Jun amino-terminal kinase（JNK） signal pathway
Abstract：ObjectiveThe effects of Atractylodes lancea， A. coreana， A. japonica， A. chinensis and Atractylodis Macrocephalae Rhizoma on spleen-Qi deficiency rats were compared.MethodA model of spleen-Qi deficiency was induced in rats by diet and overwork.The rats are given different suspensions of A.japonica， A.chinesis， A.coreana， A.lancea and Atractylodis Macrocephalae Rhizoma To test the indicators of the digestive system， immune system and antioxidant enzyme system related to spleen deficiency.Compare the similarities and differences between Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma from four different sources.ResultAll the drug-administered groups can increase the levels of γ interferon （IFN-γ）， gastric secrete element（GAS），serum amylase （AMS） and catalase（CAT） in rats with spleen-deficiency syndrome，in addition to the CAT index， the other indicators of the A. coreana， A.japonica， and A.lancea were significantly increased（P<0.05）. The MTL content of the A.chinesis， A.lancea， A.coreana and A.japonica increased and was significant（P<0.05）. The SDH content of A.japonica.and A.chinesis increased， and the difference was not significant.The increase of GSH-Px in the A.chinesis is significant（P<0.05）. All the drug-administered groups can reduce the content of IgG， TNF-α and MDA in rats with spleen deficiency and deficiency syndrome. Among them， the IgG content of the A.chinesis. and the A.lancea was significantly decreased（P<0.05）. The content of TNF-α in A.japonica group was significantly decreased（P<0.05）. The content of MDA in the A.chinesis， the A.lancea， the A.coreana，the A.japonica and the Atractylodis Macrocephalae Rhizoma were significantly decreased（P<0.05）.The decrease of IL-6 in the A.lancea was significant（P<0.05）.ConclusionFour different sources of Atractylodes Rhizome and A.macrocephala have certain therapeutic effects on spleen-deficiency rats with deficiency syndrome.The therapeutic effect of A.lancea and A.japonica is basically the same，regulating the absorption，secretion and elimination of inflammation in the digestive system of rats with spleen deficiency A.coreana， A.chinesis， and Atractylodis Macrocephalae Rhizoma have certain regulatory effects in the digestive system， digestive tract inflammation， and antioxidant enzymes.
Abstract：ObjectiveTo observe the effect of Zengmian Yiliu （ZMYL） formula and its effective components （PAWU） on the growth inhibition of ovarian cancer stem cell transplanted tumor in nude mice and the Notch signal receptor （Notch） / Notch signal ligand 1 （Jagged1） signal pathway in tumor tissue.MethodOvarian cancer stem cells were cultured in serum-free suspension to establish the transplanted tumor model of ovarian cancer stem cells in nude mice， and then divided into model group， ZMYL group （36 g·kg-1）， PAWU group （5.8 g·kg-1）， cisplatin （DDP） group （2.5 g·kg-1）， and PAWU （5.8 g·kg-1） + DDP group （2.5 mg·kg-1）.After successful modeling， the drugs were given by gavage for 21 days.To observe the effect of Zengmian Yiliu decoction and its effective components on tumor weight in nude mice， the morphological changes of tumor cells were observed under light microscope， immunohistochemistry and real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）were used to detect the expressions of Notch 1， Jagged1， Hairy and enhancer of split 1 （Hes1） protein and mRNA in tumor tissues.ResultThe tumor inhibition rates of ZMYL， PAWU， DDP and combination groups were 35.91%， 32.94%， 57.65% and 69.05%， respectively.Compared with the model group， the tumor weight of ZMYL group， PAWU group， DDP group and combination groups decreased significantly （P<0.05）.Compared with PAWU group， the tumor weight of combination groups decreased significantly （P<0.05）.Immunohistochemistry showed that compared with the model group， the positive expressions of Notch1， Jagged1 in ZMYL group， PAWU group， DDP group and combination groups were down-regulated （P<0.05），and the positive expressions of Hes1 in ZMYL group， DDP group and combination groups were down-regulated （P<0.05）.Compared with combination groups， the positive expressions of Notch1， Jagged1 and Hes1 in ZMYL group， PAWU group， DDP group were up-regulated （P<0.05）. Real time PCR showed that compared with the model group， the expressions of Notch1 mRNA in ZMYL group， PAWU group， DDP group and combination groups decreased significantly （P<0.05）.Compared with model group， the expressions of Jagged1 and Hes1 mRNA in ZMYL group， PAWU group and combination groups decreased significantly （P<0.05）.Compared with DDP group， the expressions of Notch1， Jagged1 and Hes1 mRNA in combination groups decreased significantly （P<0.05）.ConclusionThe growth of ovarian cancer stem cells transplanted in nude mice can be inhibited by Zengmian Yiliu formula and its effective components.The effective components have a significant synergistic effect in the combination with cisplatin.Its mechanism is correlated to the inhibition of Notch/Jagged1 signaling pathway activation.
Keywords：Zengmian Yiliu formula;effective components;nude mice;ovarian cancer stem cells;Notch/Jagged1
Abstract：ObjectiveThe effects of anemoside B4 on endometritis rats were studied through in vivo and in vitro experiments.MethodAnimal experiments used 25% phenol glue to prepare endometritis models. 60 female SD rats were randomly divided into blank group， model group， Kushen gel group（0.005 g·kg-1），anemoside B4 gel low，medium and high dose groups（0.005，0.01，0.02 g·kg-1），10 rats in each group，except for the blank group，rats in each group were injected with 25% phenol glue into their vagina every 2 days，and the modeling continued for 30 days. Administration started on the day after modeling. Anemoside B4 gel low， medium and high dose groups were administered rectal daily，Kushen gel group was given daily vaginal administration. The blank group and model group were given the same amount of normal saline in the same way for 30 consecutive days. After the last administration，the uterus and its attachments of each group of rats were taken to analyze the uterine morphology and index. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of rat uterus. Real-time PCR was used to detect tumor necrosis factor-α （TNF-α），interleukin-1β （IL-1β），and interleukin-6 （IL-6），signal transduction protein 130 （gp130），signal transduction and transcription activator 3 （STAT3）mRNA expression. Detection of IL-6 and STAT3 protein expression in rat uterus by Western blot. In cell experiments，lipopolysaccharide （LPS）was used to induce rat endometrial epithelial cells to prepare an in vitro inflammation model， and Real-time PCR was used to detect the expression of IL-6，gp130 and STAT3 mRNA in each group of rat endometrial epithelial cells.ResultThe results of animal experiments showed that compared with the blank group， the model group had inadequate uterine cavity adhesions， endometrial edema and hyperemia. Compared with model group， there was no adhesion in the uterine cavity of the rats in the high dose anemoside B4 gel group and the Kushen gel group. The uterine tissue was relatively complete， and the uterine pathological structure was significantly improved. Compared with the blank group， the uterine index of the model group was significantly increased（P<0.05）， the expression of IL-1β mRNA in the uterine tissue was significantly increased （P<0.05）， the expression of mRNA and protein of IL-6 and STAT3 in the uterine tissue significantly increased （P<0.05）. Compared with model group， the uterine index in anemoside B4 gel high dose group was significantly reduced （P<0.05）， and the mRNA and protein expression levels of IL-6 and STAT3 in the uterine tissue were significantly reduced （P<0.05）. There was no statistically significant difference between the TNF-α and IL-1β mRNA expression compared with the model group. Cell experiment results showed that compared with the blank group， the mRNA expression of IL-6 and gp130 in model group endometrial epithelial cells was significantly increased （P<0.01）， STAT3 mRNA expression was significantly increased （P<0.05）. Compared with model group， the mRNA expression levels of IL-6， gp130 and STAT3 in anemoside B4 high dose group decreased significantly （P<0.05）.ConclusionAnemoside B4 can improve the inflammatory response of chronic endometritis in rats and reduce the release of inflammatory factor IL-6. The mechanism may be related to the down-regulation of IL-6/STAT3 pathway.
Keywords：endometritis;anemoside B4;endometrial epithelial cells;interleukin-6（IL-6）/signal transduction and transcription activator 3（STAT3）;inflammatory response
Abstract：ObjectiveTo observe the analgesic effect of Panlongqi tablet（PLQT） on rats with chronic inflammatory pain， and to explore mechanism of the action preliminarily from the perspective of the nuclear factor kappa-light-chain-enhancer of activated B cells （NF-κB）and mitogen-activated protein kinase（MAPKs） signaling pathways.MethodRats were induced to establish model of chronic inflammatory pain by complete Freund adjuvant（CFA）， which was divided into normal group， model group， the PLQT 0.16，0.32，0.64 g·kg-1 group， and the ibuprofen 0.05 g·kg-1 group（also positive group）， give the medicine once a day by gavage. Standard Von Frey fiber was used to evaluated the mechanical pain threshold， acetone was used to stimulated rats inflammatory foot to get the cold-induced response score， with the mechanical pain threshold and cold-induced response score to be observed at 1， 2， 3， 4 and 6 h before and after administration on day 1， and at 4 h after administration on day 3-7. The content of PGE2， IL-1， TNF-α in serum， inflammatory foot and 4-5 lumbar spinal cord was detected by enzyme-linked immunosorbent assay（ELISA）. The protein level of MAPKs （p-p38， p-ERK， p-JNK） in lumbar spinal cord 4-5 was detected by Western blot. The expression of NF-κB p65 in the lumbar spinal cord was detected by IFA.ResultModel group had lower mechanical pain threshold and higher cold-induced response score than these in normal group（P<0.01）， while the mechanical pain threshold and cold-induce response score of the model rats were dose-dependent better regulated after administration of PLQT 0.16， 0.32， 0.64 g·kg-1·d-1（P<0.05，P<0.01）， these effect lasted 6 h， of which PLQT groups get the most significant effect on 4 h， however the effect of IBP was similar to that of PLQT medium dose group. In addition， PLQT reduced the abnormal increase of PGE2， IL-1 and TNF-α contents in serum， inflammatory foot and spinal cord of rats in model group， decreased the protein phosphorylation levels of ERK and JNK in spinal cord， and decreased the protein expression of NF-κB p65， that was significant in the PLQT high-dose group（P<0.01）.ConclusionPLQT had significant analgesic effect on chronic inflammatory pain model rats， which may be related to the inhibition of NF-κB and MAPKs signaling pathways in spinal cord.
Keywords：Panlongqi tablet;chronic inflammatory pain;complete freund adjuvant;nuclear factor kappa-light-chain-enhancer of activated B cells;mitogen-activated protein kinase
Abstract：ObjectiveThe methods of network pharmacology were adopted to predict Multi-component Chinese Medicine （MCCM） with anti-tumor activity from Shuanghuanglian （SHL）. Furthermore， the pharmaceutical activity of CT26 colon cancer was verified in vitro and in vivo.MethodBased on the Traditional Chinese Medicine Systems Pharmacology Database （TCMSP）， the Handbook of Active Components Analysis of Traditional Chinese Medicine， the Drugbank database， the Reactome database， and the Human protein reference database （HPRD）， the protein-protein interactions network （PPI） and the drug target network were built and resolved. The data was mined to discover the pharmacological effect. The anti- tumor activity of components from SHL was determined based on the nearest distance rule between the compounds and the nodes of network. And then， the anti-tumor effect of the MCCM was verified in vitro and in vivo.ResultThe 3 combined compounds， baicalin， forsythoside A and chlorogenic acid with the anti-tumor activity from SHL were predicted and discovered. The verification results showed that the combination of baicalin-forsythoside A-chlorogenic acid could significantly inhibit the cell proliferation and migration compared with the control group in vitro （P<0.01）. Among CT26 bearing mice， the tumor volume and weight were significantly decreased after the combined administration of baicalin-forsythoside A-chlorogenic acid compared with the model group in vivo （P<0.01）.ConclusionBy the methods of network pharmacology， the anti-tumor activities of component of from SHL were discovered. According to the verification in vitro and in vivo， the combination of baicalin-forsythoside A-chlorogenic acid could play better anti-CT26 tumor activity.
Keywords：Shuanghuanglian;multi-component Chinese medicine （MCCM）;anti-tumor;colon cancer
Abstract：ObjectiveTo investigate the pharmaceutical idiosyncratic hepatotoxicity effect of Xanthii Fructus on the immune-sensitive rat model induced by endotoxin lipopolysaccharide （LPS）.MethodThe SD rats were randomly divided into five groups： control group， model group， and three Xanthii Fructucs groups. The immune-sensitive rat model was established by LPS （iv. 0.7 mg·kg-1， twice every 7 days）. Then， the rats in control and model groups received the equal volume of distilled water， while the rats in Xanthii Fructus groups were administrated with water extract of Xanthii Fructus intragastrically （1.67， 5.01， 16.7 g·kg-1， respectively） for 14 days. The serum and liver of the rats were collected on the 7th and 14th day to examine the levels of hepatotoxic biomarkers， including alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBil）， alkaline phosphatase （ALP）， and total bile acids （TBA）， and liver histopathology. In addition， inflammatory factors， including interleukin-1β（IL-1β）， interleukin-2（IL-2）， interleukin-6（IL-6）， interleukin-10（IL-10）and tumor necrosis factor-α（TNF-α）of the idiosyncratic hepatotoxicity rats， were determined by enzyme-linked immunosorbent assay（ELISA）.ResultThe immune-sensitive model rats showed elevated levels of IL-1β， IL-6（P<0.05，P<0.01）， and mild inflammatory cells infiltrated in portal area of liver significantly （P<0.05）， with no significant changes in hepatotoxic biomarkers. Meanwhile， there was no significant change between Xanthii Fructus groups and model rats in the levels of hepatotoxic biomarkers， inflammatory factors and hepatic lesions.ConclusionWater extract of Xanthii Fructus intragastrically does not affect the levels of hepatotoxic biomarkers， inflammatory factors and hepatic lesions in rats induced by LPS intravenously. That is to say， Xanthii Fructus does not induce pharmaceutical idiosyncratic hepatotoxicity.
Abstract：ObjectiveTo analyze the common active ingredients， potential target genes and pathways of Ginseng Radix et Rhizoma "Tonifying Qi" and Notoginseng Radix et Rhizoma "Enriching blood" in alleviating fatigue based on the network pharmacology technology. And the compound ingredients of total Ginsenoside Ginseng Root and Notoginseng total Saponins were selected to verify the core target genes in vitro.MethodThe main active ingredients and related targets of Ginseng Radix et Rhizoma and Notoginseng Radix et Rhizoma were screened by traditional Chinese medicine systems pharmacology （TCMSP）. The data of fatigue genes were established by GeneCards comprehensive database and Human Mendelian Genetic Integrated Database（OMIM）. Depending， The data sets of fatigue-related genes are established based on the data bank of GeneCards and OMIM. The intersecting genes of drugs and disease were obtained by R software. Cytoscape software was used to establish the regulatory network among the active ingredients， drug targets and fatigue-related genes. PPI network of intersecting genes was constructed by STRING 11.0 software， and the core genes were screened by CytoHubba software and Matthews correlation coefficient （MCC） algorithm. Based on the results of network analysis， 24 male SPF ACR mice were randomly divided into control group， total Ginsenoside Ginseng Root group （0.08 g·kg-1） and Notoginseng total Saponins group （0.08 g·kg-1）. The corresponding drugs were given for 3 weeks. The expressions of core genes in muscle tissue were detected by real-time fluorescence quantitative PCR.ResultThe 20 active components and 181 drug targets were screened from TCMSP. 33 intersecting genes of diseases and drugs were obtained when compared with GeneCards and OMIM comprehensive database using R software. 10 core genes including aryl hydrocarbon receptor （AHR）， androgen receptor （AR）， glutathione S-transferase P1 （GSTP1）， cysteine proteinase-3（Caspase-3）， cytochrome p450 enzyme 3A4 （CYP3A4）， intercellular adhesion molecule 1 （ICAM1） and nuclear factor kappa B inhibitor alpha （NFKBIA） were screened out by the algorithm of MCC. Total Ginsenoside Ginseng Root and Notoginseng total Saponins had no significant effect on GSTP1 and ICAM1 genes， but they could significantly inhibit the expressions of AHR， CYP3A4， Caspase-3， NFKBIA and AR （P<0.05，P<0.01）， and there were no significant difference in anti-fatigue effect between total Ginsenoside Ginseng Root and Notoginseng total Saponins groups.ConclusionThe mechanism of anti-fatigue of Ginseng Radix et Rhizoma and Notoginseng Radix et Rhizoma may be related to the regulation of AHR， CYP3A4 and Caspase-3 genes， and there is no significant difference in their anti-fatigue effects， through the analysis of network and experimental verification.
Keywords：Notoginseng total saponins;network pharmacology;Notoginseng Radix et Rhizoma;Ginseng Radix et Rhizoma;total Ginsenoside Ginseng Root;fatigue syndrome
Abstract：ObjectiveTo investigate the clinical efficacy of Loulu Shengma Tang combined with azithromycin in the treatment of pediatric mycoplasma pneumoniae pneumonia（MPP） with obstruction of lung by pathogenic heat， and its effect on inflammatory factors， treg and Foxp3 mRNA.MethodTotally 274 children with MPP were divided into observation group （137 cases） and control group （137 cases）. Observation group was treated with Loulu Shengma Tang combined with azithromycin dry suspension， while control group was treated with azithromycin dry suspension alone. The changes of traditional Chinese medicine（TCM） syndrome score of pathogenic-heat obstruction in the lung， serum inflammatory cytokines ［interleukin-6 （IL-6），interleukin-10 （IL-10），tumor necrosis factor-α （TNF-α），C-reactive protein （CRP）］， CD4+CD25+Treg， CD4+Foxp3+Treg and Foxp3 mRNA expressions were observed after treatment. The clinical efficacy and the incidence of adverse reactions were compared between two groups.ResultThe total effective rate of observation group was 94.16%（129/137） after treatment， which was significantly higher than 77.37% （106/137）of observation group （P<0.05）. The disappearance times of cough， lung rale， fever and lung shadow in observation group were shorter than that in control group （P<0.05）. After treatment， TCM syndrome score of pathogenic-heat obstruction in lung was significantly higher than those in control group （P<0.05）， serum IL-6， IL-10， TNF-α and CRP levels in observation group were significantly lower than those in control group （P<0.05， P<0.01）， while CD4+CD25+Treg， CD4+Foxp3+Treg and Foxp3 mRNA expressions were significantly higher than those in control group （P<0.05）. The incidence of adverse reactions in observation group was 7/137 （5.11%）， which was significantly lower than 16/137 （11.68%） in control group.ConclusionThe clinical efficacy of Loulu Shengma Tang combined with azithromycin dry suspension in the treatment of pediatric MPP and its effect on serum inflammatory factors （IL-6， IL-10， TNF-α， CRP）， regulatory T cells and Foxp3 mRNA expressions were better than those of azithromycin dry suspension alone. The incidence of adverse reactions of Loulu Shengma Tang was lower than that of azithromycin dry suspension alone.
Keywords：Loulu Shengma Tang;azithromycin;mycoplasma pneumonia in children;pathogenic-heat obstruction in the lung;regulatory T cells;Foxp3 mRNA
Abstract：ObjectiveTo explore the efficacy of Liuwei Nengxiao capsule combined with fecal bacteria transplantation on chronic functional constipation with spleen Qi deficiency.MethodA total of 129 patients with chronic functional constipation of spleen Qi deficiency treated in Affiliated Hospital of Shandong University of Traditional Chinese Medicine were randomly divided into three groups by the computer numerical table method： fecal bacteria transplantation group， Liuwei Nengxiao capsule group， and Liuwei Nengxiao capsule combined with fecal bacteria transplantation group， with 43 cases in each group. Fecal bacteria transplantation group was treated with fecal bacteria， Liuwei Nengxiao capsule group was treated with Liuwei Nengxiao capsule， and Liuwei Nengxiao capsule combined with fecal bacteria transplanted group was treated with Liuwei Nengxiao capsule combined with fecal bacteria. The efficacy of the 3 groups was compared.ResultAfter treatment， the frequency and amplitude of intestinal electricity increased significantly in three parts， with statistically significant differences from before treatment （P<0.05）. After treatment， the frequency and amplitude of the transverse colon， sigmoid colon， and descending colon of patients in Liuwei Nengxiao capsule combined with faecal transplantation group were higher than those in faecal transplantation group and Liuwei Nengxiao capsule group （P<0.05）. After treatment， substance P （SP） and motilin （MTL） of Liuwei Nengxiao capsule combined with faecal transplantation group were higher than those of faecal transplantation group and Liuwei Nengxiao capsule group， while nitric oxide （NO） and vasoactive intestinal peptide （VIP） were lower than those of faecal transplantation group and Liuwei Nengxiao capsule group （P<0.05）. The number of bifidobacteria and lactobacillus after treatment was higher in three groups than before treatment （P<0.05）. The numbers of yeast and enterobacteria were lower than before treatment （P<0.05）. After treatment， the number of bifidobacteria and lactobacillus in Liuwei Nengxiao capsule combined with faecal transplantation group was higher than that in faecal transplantation group and Liuwei Nengxiao capsule group， whereas the numbers of yeast， and enterobacteria were all lower than those in bacterial transplantation group and Liuwei Nengxiao capsule group （P<0.05）. After treatment， the BSFS scores of three groups were higher than before the treatment， while Wexner continence grading scale （Wexner） constipation score and patient assessment of constipation quality of life questionnaire （PAC-QOL） score were lower than before treatment （P<0.05）. Bristol stool form scale （BSFS） score of Liuwei Nengxiao capsule combined with fecal bacteria transplantation group was higher than that of fecal bacteria transplantation group and Liuwei Nengxiao capsule group. Wexner constipation score and PAC-QOL score were lower than those of fecal bacteria transplantation group and Liuwei Nengxiao capsule group （P<0.05）.ConclusionFecal bacteria transplantation combined with Liuwei Nengxiao capsule can effectively promote the recovery of intestinal electrical function， improve the intestinal flora balance， reduce the intestinal oxidative stress response， and promote the disappearance of patients' symptoms in the treatment of elderly chronic functional constipation， and thus is worth further promotion in clinic application.
Abstract：ObjectiveTo discuss influence of addition and subtraction therapy of Zhenwutang to residual renal function （RRF）， nutritional status， dialysis adequacy and quality of life of patients with maintenance hemodialysis （MHD）.MethodOne hundred and thirty-six patients were randomly divided into control group （68 cases） and observation group （68 cases） by random number table. Patients in two group got MHD， 3 times/week， 4 h/time， levocarnitine injection （1 g dissolved in 5-10 mL water for injection） after the dialysis， 2-3 min/time， recombinant human erythropoietin injection with subcutaneous injection for 4 weeks， 3 000 U， 3 times/day， valsartan capsules for 3 months， 80 mg/time， 1 time/day. The control group took Manshenning mixture， 35 mL / time， 3 times / day.Patients in observation group added addition and subtraction therapy of Zhenwutang for 3 months， 1 dose/day. Before and after treatment， urea nitrogen （BUN）， creatinine （CR） and 24 hours' urine volume were recorded. And RRF， rate of decrease of RRF and rate of decrease in urine volume were also calculated. Levels of hemoglobin （HB）， albumin （ALB）， prealbumin （PA） and transferrin （TRF） were detected. After treatment， standardized protein metabolism rate （nPCR）， urea clearance index （Kt / V） and glomerular filtration rate （EGFR） were discussed. And improved subjective comprehensive nutrition assessment （SGA）， dialysis related quality of life （kdta） and health survey summary （SF-36） were graded. Six months' follow-up， primary end point event （24 h urine volume ≤ 400 mL） and no residual renal function （24 h urine volume ≤ 400 mL） were recorded.ResultLevels of RRF， Kt/V， nPCR， eGFR， Hb， Alb， PA， TRF and total scores of KDTA and SF-36 in observation group were higher than those in control group （P<0.01）. And score of SGA， rate of decrease of RRF and rate of decrease in urine volume were less than those in control group （P<0.01）.Incidence rate of primary end point event was 27.94%（19/68） lower than 47.06%（32/68） in control group （χ2=5.302， P<0.05）， incidence rate of no residual renal functionwas 17.65%（11/68） lower than 36.76%（25/68） in control group （χ2=6.274， P<0.05）. And BUN and Cr were lower than those in control group （P<0.01）， 24 h urine volume was more than that in control group （P<0.01）.ConclusionAddition and subtraction therapy of Zhenwutang can maintenance of RRF， improvement of nutritional status， improvement of dialysis adequacy and quality of life of patients .
Keywords：maintenance hemodialysis;Zhenwutang;residual renal function;nutritional status;quality of life
Abstract：ObjectiveTo evaluate the clinical efficacy of Sanjiao Cidi therapy on acute cerebral infarction and its effect on levels of S100-β protein （S100-β）， tumor necrosis factor-α （TNF-α）， hypersensitive C-reactive protein （hs-CRP） and neuropeptide （NPY）.MethodOne hundred and eighty patients were randomly divided into control group （90 cases） and observation group （90 cases） by random number table. Patients in control group got aspirin enteric-coated tablets， 100 mg/time， 1 time/day， edaravone injection （injected within 30 minutes） for 14 days， 30 mg/time， 2 times/day， simvastatin tablets， 20 mg/time， 1 time/day. In addition to the basic therapy of meloxicam tablets， patients in observation group were also treated with Sanjiao Cidi therapy. In the first step， patients got Guizhi therapy to dredge Zhongjiao and Shangjiao， 1 dose/day， for 8 days. In the second step， patients got Sini therapy to dredge Zhongjiao and Xiajiao， 1 dose/day， for 10 days. In the third step， patients got Tianjing Gubentherapy， 1 dose/day， for 10 days. The course of treatment was 4 weeks. Before the treatment， and at the first， second， third and fourth weeks after treatment， National Institutes of Health Stroke Scale （NIHSS） was scored. And before and after treatment， function scale of fuglmeyer （FMA）， ability of daily life activities （ADL）， mini-mental state examination （MMSE） and main symptoms of traditional Chinese medicine were scored. Comprehensive assessment of patient report outcome （PRO） was made. And levels of S100-β， hs-CRP， TNF-α and NPY were detected. And the incidence rate of pulmonary infection， urinary infection， skeletal myalgia， shoulder hand syndrome and shoulder subluxation of patients were recorded during hospitalization.ResultThe clinical efficacy in observation group was better than that in control group （Z=2.141， P<0.05）. Scores of NIHSS in observation group were lower than those in control group at the first， second， third and fourth weeks after treatment （P<0.01）. Scores of upper limb， legs and the total scores from FMA were higher than those in control group （P<0.01）. Scores of the main symptoms of traditional Chinese medicine， symptoms， psychological and social scores， total scores of PRO， S100-β， hs-CRP， TNF-α and NPY were lower than those in the observation group （P<0.01）. And scores of ADL and MMSE were higher than those in control group （P<0.01）. Total incidence of complications in observation group was 27.27%（21/77）， which was lower than 46.15%（36/78） in control group （χ2=5.941， P<0.05）.ConclusionIn addition to conventional western medicine treatment， Sanjiao Cidi therapy can treat the patients with acute cerebral infarction， alleviate the degree of neurological deficit， improve the cognitive function， motor function of limbs and the ability of daily life， reduce the main symptoms of traditional Chinese medicine， the incidence of complications and the inflammatory response， protect the nerve cells， with a better clinical efficacy and comprehensive effect in patients than pure Western medicine.
Keywords：cute cerebral infarction;triple focus therapy;Fuyang theory;Guizhitang;Sinitang;Fuzitang;nerve function defect
Abstract：ObjectiveTo evaluate the regulation effect of corn silk on fatty acid compositions in adipose tissues of rats with type 2 diabetes mellitus（T2DM） based on gas chromatography-mass spectrometry （GC-MS）， and explore the mechanism of corn silk water decoction for lipid metabolism disorder in .MethodRat model of T2DM was established by high-sugar and high-fat diet combined with intraperitoneal injection of 35 mg·kg-1 streptozocin （STZ）. They were randomly divided into model group，metformin group，and low，medium and high dose groups（5.4，10.8，21.6 g·kg-1） of corn silk decoction， with administration of 4 weeks. Serum was taken from the abdominal aorta for the determination of free fatty acids （FFAs），blood lipids ［total cholesterol（TC），triglyceride（TG），high-density lipoprotein（HDL-C），low density lipoprotein（LDL-C）］，and adipose tissue was taken for pathological section analysis. At the same time，GC-MS technology and multivariate statistical analysis methods were adopted to investigate the fatty acid compositions in adipose tissue.ResultAfter treatment with corn silk decoction，the contents of FFAs，TC，TG，HDLC and LDLC in T2DM rats were all reversed. As compared with the model group，the morphology of rat adipocytes，macrophage infiltration，vascular congestion and other phenomena were restored after administration. GC-MS analysis showed that the contents of palmitic acid and stearic acid in the adipose tissue of T2DM rats were significantly increased，while the contents of tetradecanoic acid，pentadecanoic acid，9-octadecenoic acid，9-hexadecenoic acid and 5，8，11，14-eicosatetraenoic acid were decreased significantly，and all the dose groups of corn silk had a reversing effect on differentially expressed fatty acids.ConclusionCorn silk can be able to reverse the fatty acid content in adipose tissue and restore the morphology of adipose tissue，with a regulating effect on the lipid metabolism disorder in T2DM rats.
Abstract：ObjectiveThere were 92 kinds of compound preparations containing Ophiopogonis Radix in the 2015 edition of Chinese Pharmacopoeia， but there was no effective method to identify these compound preparations. Because Ophiopogonis Radix and Liriopes Radix are similar in appearance， it is easy to be confused in application. The aim of this study was to set up a thin layer chromatography （TLC） to identify compound preparations containing Ophiopogonis Radix and distinguish Ophiopogonis Radix and Liriopes Radix in the forms of decoction pieces and standard decoction.MethodIn this study， decoction pieces of Ophiopogonis Radix and Liriopes Radix were collected and separately prepared as standard decoction. TLC was used to qualitatively identify decoction pieces and standard decoction of Ophiopogonis Radix and Liriopes Radix， and compound preparations containing Ophiopogonis Radix. In the TLC， the lower solution of chloroform-methanol-water （65∶35∶10） was selected as the developing agent and 10% sulfuric acid ethanol solution as the chromogenic agent.ResultThe resolution of this TLC was good. Decoction pieces， standard decoction and preparations of Ophiopogonis Radix had the same characteristic strips， which were two bright white fluorescent strips under ultraviolet lamp （365 nm）. But these two characteristic strips were not existed in the TLC of decoction pieces and standard decoction of Liriopes Radix. The corresponding components of both of these two strips were identified as mixture containing saponins by LC-MSn， including ophiopogonin Ra， Tb， ophiopogonin D'， borneol glycoside， ophiopogonin C and Liriope muscari baily saponins C.ConclusionThe established TLC method， which has significant advantages such as high specificity and sensitivity， can be applied to the characteristic identification of decoction pieces and standard decoction of Ophiopogonis Radix， the identification of compound preparations containing Ophiopogonis Radix， and the distinction of Ophiopogonis Radix and Liriopes Radix， thus serving as an effective method to qualitatively identify Ophiopogonis Radix and its compound preparations.
Abstract：ObjectiveTo improve the quality standard of Shenwei Gubi tablets， and to explore the reasons for the great difference in the contents of quality control index components between batches of this product.MethodThe fingerprint of this product was established by HPLC， the determination was performed on Diamonsil C18 column （4.6 mm×250 mm， 5 μm） with acetonitrile （A）-0.1% phosphoric acid solution （B） for gradient elution （0-5 min， 10%A； 5-15 min， 10%-12%A； 15-30 min， 12%-26%A； 30-43 min， 26%-31%A， 43-50 min， 31%-40%A， 50-70 min， 40%-55%A； 70-84 min， 55%-72.5%A） as the mobile phase at detection wavelength of 230 nm. The orthogonal partial least squares-discriminant analysis-variable importance in the projection （OPLS-DA-VIP） map was drawn with the common peak as the independent variable. The contribution of 26 common peaks to the fingerprint differences among different batches of this product was quantified. By searching for the chromatographic peaks with great differences， combined with relevant literature， the components related to the clinical indications of the product were screened out and their contents were determined by specificity experiment， and the quantitative indicators were finally selected. HPLC-doide array detector （DAD） was employed to determine the contents of the above preferred indexes with detection wavelengths of 236， 276， 230， 322 nm， other conditions were the same as HPLC fingerprint detection method.ResultA total of 26 common peaks were calibrated on the HPLC fingerprint of Shenwei Gubi tablets. The similarity between the fingerprint of each batch samples and the reference fingerprint was≥0.950. Loganic acid， gentiopicroside， paeoniflorin and osthole were optimized as the quantitative indicators of this product， their average contents were 161.02， 401.80， 255.54， 80.68 μg·g-1.ConclusionThe established fingerprint and multi-index quantitative analysis method are stable and reliable， and can be used for quality control of Shenwei Gubi tablets. Difference in contents of quality control components between batches of raw materials and the imperfect quality control method of intermediates in the production process are the main reasons for the great difference in the contents of quality control indicators between batches of this product.
Keywords：fingerprint;chemical pattern recognition;cluster analysis;principal component analysis （PCA）;orthogonal partial least squares-discriminant analysis （OPLS-DA）;high performance liquid chromatography （HPLC）;Shenwei Gubi tablets
Abstract：ObjectiveTo investigate the effect of different angles of atomizer on the delivery rates and total delivery quantities of Tanreqing inhalation solution， so as to provide reference for the clinical use of this preparation.MethodTaking baicalin， ursodeoxycholic acid， caffeic acid as indexes， PARI Boy SX compression atomization inhaler （equipped with red core atomizing cup） and BRS2000 respiratory simulator were used， the effects of different angles of the atomizer （upper 15 degree， lower 15 degree， upper 30 degree， lower 30 degree， partial 15 degree， partial 30 degree， vertical） on the delivery rates and total delivery quantities of Tanreqing inhalation solution were investigated. The respiratory patterns of adults， children， infants and young children were selected to determine the delivery rates and total delivery quantities of three components in Tanreqing inhalation solution.ResultIn the same atomization time， the delivery rates and the total delivery quantities of caffeic acid and ursodeoxycholic acid in Tanreqing inhalation solution were not significantly affected by the atomizer from different angles， but significantly affected baicalin. At the vertical angle， the delivery rate and total delivery quantity of baicalin were higher than the other angles. Under different respiratory modes， there were significant differences in the delivery rates and total delivery quantities of these three components in the inhalation solution. Compared with other respiratory modes， the delivery rates and total delivery quantities of baicalin， ursodeoxycholic acid and caffeic acid were the highest in the adult respiratory mode， with delivery rates of （555.5±16.61）， （226.3±6.54）， （26.1±0.32） μg·min-1 and total delivery quantities of （4 001.1±82.97）， （1 754.9±63.73）， （167.6±1.42） μg， respectively.ConclusionThe use angle of atomizer has a certain effect on the delivery rate and total delivery quantity of Tanreqing inhalation solution， so it is suggested that the vertical angle should be kept as far as possible in clinical use. Under the four respiratory patterns， the delivery rate and total delivery quantity of Tanreqing inhalation solution are different， suggesting that the atomization dose or atomization time should be adjusted according to the respiratory characteristics of the patients to ensure the safety and effectiveness of clinical medication.
Abstract：ObjectiveTo establish a thin layer chromatography （TLC） identification method for different processed products of Rehmanniae Radix.MethodCatalpol， D-fructose， sucrose， raffinose， stachyose， melibiose and manninotriose were used as control substances， and the effects of extraction solvents （water， 20% methanol， 50% methanol， 80% methanol）， developing solvents （n-butanol-methanol-chloroform-glacial acetic acid-water， ethyl acetate-pyridine-glacial acetic acid-water， n-butanol-glacial acetic acid-water）， color reagents （aniline-diphenylamine-phosphoric acid solution， ninhydrin solution）， sampling volumes （2， 4， 6 μL） and inspection conditions （sunlight， bottom lamp of sunlight， 365 nm， 254 nm） on TLC were investigated to determine the preparation method of sample solution of Rehmanniae Radix and Rehmanniae Radix Praeparata and the optimal TLC conditions.ResultHigh performance silica gel G plate was used for TLC， n-butanol-methanol-chloroform-glacial acetic acid-water （13∶5∶5∶1∶2） was used as developing agent， aniline-diphenylamine-phosphoric acid solution was sprayed and heated at 110 ℃ for color development， and then inspected under the bottom lamp of sunlight. The separation and color development of different processed products of Rehmanniae Radix were good with clear spots and good characteristics.ConclusionThe established TLC is simple and easy to operate with obvious qualitative characteristics and intuitive results. It can effectively identify different processed products of Rehmanniae Radix and provide experimental basis for determining the end point of processing of Rehmanniae Radix Praeparata.
Abstract：ObjectiveTo explore genetic relationship and population structure of Turpinia arguta in six locations of Jiangxi province by inter-simple sequence repeat （ISSR） molecular marker technique， and to provide theoretical basis for the protection and utilization of this medicinal material resource.MethodA total of 22 samples from six locations in four counties in Jiangxi province were collected， and genomic DNA was extracted by kit method. Polymerase chain reaction （PCR） amplification was performed using sixty-four universal ISSR molecular marker primers， and the products were detected with polyacrylamide gel electrophoresis （PAGE）. NTsys 2.10e software was selected to calculate the genetic similarity coefficient by unweighted pair group method with arithmetic mean （UPGMA） and cluster analysis. Population genetic structure was analyzed by Structure 2.1 software.ResultA total of forty-eight ISSR primers were amplified to obtain the product， the percent of polymorphic bands ranged from 45.45% to 100%. UPGMA cluster analysis showed that these plant individuals could not be clustered according to their respective executive locations. Analysis of population genetic structure showed that 22 samples of T. arguta could be divided into three populations.ConclusionThere is gene exchange among the populations of T. arguta in Jiangxi province， and it can affect the genetic structure of germplasm resources from different geographical sources.
Keywords：Turpinia arguta;inter-simple sequence repeat （ISSR）;molecular marker;genetic relationship;population genetic structure;polyacrylamide gel electrophoresis （PAGE）;unweighted pair group method with arithmetic mean （UPGMA）
Abstract：ObjectiveA high performance liquid chromatography-photo-diode array（HPLC-PDA） method for the simultaneous determination of the 7 phenolic acids including danshensu，protocatechuic acid，protocatechuic aldehyde，chlorogenic acid，caffeic acid，ferulic acid and rosemary acid in Lycopus lucidus var.hirtus rhizome，analyzing and evaluating the phenolic acids in L.lucidus var.hirtus rhizome collected from different habitats，is reported here.MethodThe sample was extracted by ultrasonic with 80% methanol solution，7 kinds of phenolic acids were separated on a CAPCELL PAK C18 column （4.6 mm×250 mm，5 μm） with a mobile phase consisting of methanol-0.02% formic acid aqueous （pH 3.10）by gradient elution，The detection wavelength was at 279，324 nm， the column temperature was 30 ℃ with 20 μL injection volume and the flow rate was 1.0 mL·min-1.ResultThe 7 Phenolic acids had a good linear relationship （r≥0.999 9） within their respective mass concentration ranges，the average recovery was 96.49%-103.45% and the RSD was 0.5%-2.8%，the limit of determination was 0.008-0.046 mg·L-1 and the limit of quantification was 0.027-0.154 mg·L-1.The 7 kinds of phenolic acids were all detected in L.lucidus var.hirtus rhizome and the total amount was between 5 811.01 and 11 747.23 µg·g-1 ， the average amount was 7 421.05 µg·g-1.The content of 7 phenolic acids was different and the rosemary acid was the highest in all the samples with an average of 7 111.19 µg·g-1 the ratio to the total phenolic acids was 95.8%.The results of principal component analysis and cluster analysis showed that the quality of L.lucidus var.hirtus rhizome from Heze city in Shandong province was better，followed by Wanzhou district in Chongqing.ConclusionThe method was simple，sensitive，accurate，practical and reliable，and is suitable for the content determination of phenolic acid in L. lucidus var. hirtus rhizome.It is expected to provide a reference for the improvement of quality standard and a new idea for the development and utilization of L.lucidus var.hirtus rhizome.
Abstract：ObjectiveTo systematically investigate the chemical constituents of 90% ethanol extract of Corydalis impatiens，and evaluate their inhibitory effect on cell proliferation in vitro.MethodThe chemical constituents were isolated and purified by silica gel，Sephadex LH-20，ODS column chromatography and semi-preparative high performance liquid chromatography （HPLC）.the structures were identified by spectroscopy methods such as NMR and MS，as well as analysis of physicochemical properties and/or comparison with literature data，and the inhibitory activities of 13 compounds on HepG2 and SMMC-7721 cells were measured by methyl thiazolyl tetrazolium （MTT） method.ResultThirteen known compounds were isolated and identified from 90% ethanol extract of Corydalis impatiens as ethyl-5-hydroxy-2-pyridinecarboxylate（1），coryhumolide（2），3，4-cis-3，4-dihydroxy-β-ionone（3），megastigmane（4），9-hydroxy-4，7-megastigmadien-3-one（5），blumenol A（6），indole-3-carboxy acid（7），1-methyl-[1，2，4]triazolo[4，3-b][1，2，4]triazin-7-one （8），adenine（9），nicotinamide（10），2-hydroxymethyl-5-pyridinol（11），adenosine（12）， and β-daucosterol（13）. Cytotoxicities assay showed that the IC50 value of compound 3 for the hepatic cell line HepG2 was 24.7 μmol·L-1 （positive control drug cisplatin：4.8 μmol·L-1），and IC50 value of compound 4 for the hepatic cell line SMMC-7721 was 13.8 μmol·L-1（positive control drug cisplatin：5.4 μmol·L-1）.ConclusionCompound 1 was a new natural compound，compounds 3-8 were obtained from genus Corydalis for the first time，and compounds 2，9-12 were isolated from this plant for the first time. Compound 3 exhibited weak inhibitory effect on hepatic cell line HepG2，and compound 4 exhibited moderate inhibitory effect on hepatic cell line SMMC-7721.The other compounds rest of ones exhibited no obvious inhibitory effect on hepatic cell line HepG2 or SMMC-7721.
Abstract：ObjectiveTo explore the correlation between effective constituent content and external color by measuring the chroma value and the content of polysaccharide，saponin and lovastatin during the fermentation of Polygonati Rhizoma samples， in order to provide reference for the quality control of Monascus fermented Polygonati Rhizoma.MethodPolygonati Rhizoma was inoculated with Monascus for solid-state fermentation，and the content of polysaccharide and saponin was determined by colorimetric method， while the content of lovastatin，a new ingredient after fermentation，was determined by HPLC. The chroma values L*，a*，b* of all these samples were measured by a color difference meter. SPSS 20.0 was used to analyze the correlation and regression between component content and chroma value，and the differences between samples in the fermentation process was studied through multi-dimensional scale analysis and cluster analysis.ResultWith the prolongation of fermentation time，the content of polysaccharide gradually increased to the peak value on day 9， and then gradually decreased. The content of saponin showed an rising trend in general，with slight fluctuations in the middle. Lovastatin levels peaked on day 15， and fell sharply since then. The content of polysaccharide was positively related to the lightness value L* and the total color value E*ab （P<0.01），and the contents of saponin and lovastatin were significantly related to the red-green value a*（P<0.01）. The results of regression analysis showed that 52.8% of the change in lightness value L* was affected by the content of polysaccharide，saponin and lovastatin，and the saponin content could be predicted by a* and L* to the extent of 54.1%，while the lovastatin content can be predicted by a* to the extent of 61.7%. The determination coefficient of the polysaccharide regression model was less than 0.4，so the polysaccharide content could not be predicted based on the chroma value. Through the multi-dimensional scale analysis and cluster analysis，the samples were divided into four categories according to fermentation time： 0-3，3-7，7-11，11-18 d.ConclusionThe fermentation of Monascus and Polygonati Rhizoma can lead to the change of effective constituent and appearance color of the latter，which provides the basis for the fermentation process of Polygonati Rhizoma. In addition， the content of saponin and lovastatin can be predicted by the chroma value of fermented Polygonati Rhizoma to a certain extent，which provides a simple and fast inspection method for the dynamic monitoring of effective constituent content in fermented Polygonati Rhizoma.
Abstract：This paper origin studies the origin of Alismatis Rhizoma in Chinese pharmacopoeia， and puts forward some suggestions for modification. Through the changes in the records of the source of Alismatis Rhizoma in the various versions of the Chinese Pharmacopoeia and the records of Flora of China and Materia Medica of China，it is found that the source of Alismatis Rhizoma in the Chinese Pharmacopoeia is confused. Specifically， the Chinese name of Alismatis Rhizoma does not correspond to the Latin name. As a common Chinese herbal medicine，Alismatis Rhizoma has a large market circulation. Many classic Chinese medicine prescriptions released by China Food and Drug Administration contain Alismatis Rhizoma. The development of the classic Chinese medicine prescriptions will further increase the market circulation of Alismatis Rhizoma. As a major national move to promote the development of traditional Chinese medicine， the study for classic Chinese medicine prescriptions requires defining the origin of the medicinal materials used，and the confused origin of Alismatis Rhizoma recorded in the Chinese Pharmacopoeia seriously hinder the development of the classics. Therefore，in order to regulate the origin of Alismatis Rhizoma， ensure the clinical efficacy and promote the development of classic Chinese medicine prescriptions，the confused origin of Alismatis Rhizoma in the Chinese Pharmacopoeia has to be resolved as soon as possible. Based on the analysis of the changes of Alismatis Rhizoma's producing areas in the past dynasties， it is found that the producing areas of Alismatis Rhizoma have continuous changed from Wei and Jin dynasties to present， and finally formed the current situation of Sichuan as the main producing area. In comparison of chemical composition，origin and market circulation of Alismatis Rhizoma in Sichuan Province that is the most productive， and Fujian Province that is the best quality， it is found that the two species are different in every aspects. Nowadays，Alisma plantago-aquatica occupies the majority of the market， which doesn't conform to Alisma orientale as specified in the 2015 edition of the Chinese Pharmacopoeia. Therefore， through textual research and analysis， it is suggested that both A. plantago-aquatica and A. orientale. Shall be used as the origin of Alismatis Rhizoma. In the 2015 edition of Chinese Pharmacopoeia，Cassiae Semen，Schizonepetae Herba，Aisaematis Rhizoma，Fibraureae Caulis and Ajugae Herba have the same problem. This paper provides ideas for the revision of sources of traditional Chinese medicine.
Keywords：Alismatis Rhizoma;Alisma orientale （Sam.） Juzep.;Chinese Pharmacopoeia;origin;change of origin
Abstract：The famous classical formulas （FCF）， which occupy the most important position in the prescription theory， had been brought out following the progress of traditional Chinese medicine （TCM）. Its formation and exploitation， essentially the ingredient constitution and preparation technology， is an objective trend of TCM. The development of related product under the guidance of relevant national policies has its historical inevitability， as the development of formula granules in recent decades had laid a solid foundation for it. The key to exploit these FCF is to refine and standardize the clinical applications through the ages with the guidance of TCM theory， for the process design of massive industrial production. As a result of the analysis of its definition and connotation， it is clear that the core connotation of FCF is the medicinal substances with fixed formula compatibility and stable preparation process. Furthermore， the screening strategy， the exploitation value， and the applicability between ancient prescriptions and nowadays diseases， had also been discussed based on the current policy situation and research status.
Keywords：famous classical formulas;formula theory;compound preparation;product development;standardization;process design;quality control
Abstract：Chinese patent medicines have the basic characteristics of great clinical value， strong scientific value and high market value. It is not only the core engine of traditional Chinese medicine （TCM） industry development but also the important content of TCM academic development， which is the great significance of the cause and industry of TCM. In this study， based on reviewing the methods and models of TCM theoretical， the innovation of TCM theory and Chinese patent medicines were hitting-point， which it is emphasized that the creation of effective formula is the concentrated embodiment of the theoretical innovation research results， and the Suhuang Zhike capsule and fengke theory， Tongxinluo capsule and collateral disease theory were taken as examples. On this basis， the ideas and strategies were put forward to realizing the theoretical innovation research of TCM with the large varieties of TCM as the carrier， that is， based on combing and analyzing the prescription rules and functional indications of the large varieties of TCM， and deepening the theoretical connotation of it. Meanwhile， not only emphasized the general theoretical understanding but also paid attention to the application of unique drugs and their compatibility. It not only satisfies the modern scientific connotation of revealing the role of TCM， but also explores unknown or possible new targets， new pathways and mechanisms by using large varieties of proprietary Chinese medicines to promote scientific discoveries and lead medical research.
Keywords：Chinese patent medicine;large varieties;theory of traditional Chinese medicine
Abstract：Pyroptosis，which is different from cell necrosis， apoptosis and autophagy，is a new type of programmed cell death. It is characterized by the swelling of cells until the cell membrane rupture， resulting in the release of cell contents and the activation of a strong inflammatory response. Pyroptosis signaling pathway is divided into the classical pathway dependent on Caspase-1 and the non-classical pathway dependent on Caspase-4， 5 and 11. In classical pathway， activation Caspase-1 depends on inflammasome， whereas in non-classical pathway， Caspase-4， 5， and 11 are activated directly. All of these result in the cleavage of gasdermin D（GSDMD） protein， the formation of membrane pores， the maturation and release of interleukin-1β（IL-1β） and IL-18， and the rupture of the cell membrane， and then leads to pyroptosis. Ulcerative colitis （UC） is a common disease of the digestive system. The UC course is long， easy to relapse， and difficult to cure. The pathogenesis of UC is not completely clear. In recent years， it has been found that pyroptosis plays an important role in the development of UC. Traditional Chinese medicine（TCM） has a long history of preventing and treating UC， with significant clinical effects. It is found that the mechanism of TCM in the prevention and treatment of UC is closely correlated with inflammasome， IL-1β and IL-18. The mechanism of TCM in preventing and treating UC is correlated with cell pyroptosis. Therefore， this paper reviews the relationship between cell pyroptosis and UC and the regulatory effect of TCM.
Keywords：pyroptosis;ulcerative colitis;Nod-like receptor protein 3（NLRP3）;traditional Chinese medicine
Abstract：Tic disorder （TD） is a neurodevelopmental disorder， with one or more motor and/or vocal disorders as the main symptoms. It brings many inconveniences to children's learning and life， and has a profound impact on children's character building. The pathogenesis of TD is mainly correlated with neurotransmitter release disorder， neuroimmune， genetic， trace element imbalance， diet and other factors， but has not been completely clear up to now. Western Medicine has obvious effects on TD， but with serious side effects. Compared with western medicine， traditional Chinese medicine （TCM） has the advantages of low adverse reactions and definite and lasting effect， and thus has been widely recognized by children and their families. In order to explore the pathogenesis of TD and the specific mechanism of TCM in the treatment of TD， many scholars have carried out a large number of in-depth animal experiments and made some achievements， but also exposed some defects， such as the single modeling method， failed to take into account other pathogenesis of TD， failure to combine the specific syndromes of TCM for targeted modeling， and failure to reflect the dialectic of TCM on the characteristics of governance. This paper reviews the modeling methods of common animal models， the comparison of advantages and disadvantages， and the changes of behavioral and biochemical indicators before and after the intervention with TCM compounds on TD animal models， so as to provide reference for the selection of animal models in future animal experimental research.
Keywords：tic disorder;animal model;traditional Chinese medicine;research progress
Abstract：Artemisia argyi is a perennial aromatic herb. As been recorded in 2015 edition of Chinese Pharmacopoeia， it has the characteristics of warming meridian to stop bleeding，dispersing cold and relieving pain，and clearing dampness to stop itching in topical use. It contains a variety of active ingredients and has a wide range of edible and medicinal values. It mainly includes essential oil，flavonoids，polysaccharides，tannins and other compounds. The essential oil compounds have antioxidant，anti-inflammatory，anti-tumor and bacteriostatic biological activities. At the same time，they have activities of poisoning，repelling，antifeeding and affecting growth and development for the mosquitoes. In order to further study the activity of essential oil and its chemical components from A. argyi in mosquitoes control，and to improve the development and utilization of mosquito control products，we reviewed the functions of plant essential oil for mosquito control and the existing related dosage forms in this study. Simultaneously，we summarized the extraction methods， mosquito control functions and active ingredients of essential oil， with focus on the action mode and target objects of the active components in volatile oil monomers from A. argyi. Based on the current researches on essential oil of A. argyi in mosquitoes control，the existing problems were analyzed to provide references for the synthesis and application of plant insecticides，environment-friendly pesticides and new insecticides developed as green plant materials，and put forward the prospects on the development of new highly active A. argyi mosquitoes control products.
Keywords：Artemisia argyi;essential oil;active ingredients;control insect pests and mosquitoes
Abstract：The incidence of high-normal blood pressure gradually increased. With the popularization of basic medical knowledge， the detection rate has increased. Without intervention， high-normal blood pressure is very likely converted into hypertension， and the risk of cardiovascular and cerebrovascular diseases will continue to rise. Studies have shown that the blood lipids for detection of people with high-normal blood pressure， cytokines and other laboratory indicators have been changed， causing certain damage to target organs. Normal-high blood pressure people and high-blood pressure people also need to be given attention. Traditional Chinese medicine （TCM） believes that high-normal blood pressure has intermingled deficiency and excess， which is closely correlated to the liver， spleen and kidney. The syndrome is constantly changing and developing in the course of disease， which is affected by physical fitness， environment， age and other factors. With respect to treatment， conventional western medicine for lowering blood pressure has not yet been incorporated into the guidelines， and exercise， diet， and health education still play a major role. Studies have shown that the intervention methods with TCM characteristics are applied in addition to the improvement of lifestyle， so as to intervene in people with high-normal blood pressure， such as TCM， acupuncture， herbal tea， Baduanjin， with a significant clinical effect. These therapies can effectively reduce blood pressure， improve symptoms， regulate physique， and protect target organ damage， with a good compliance. With the advantages in preventing the disease， TCM is of far-reaching significance to prevent disease in advance， and transform high-normal blood pressure into ideal blood pressure. Based on the study on the intervention of high-normal blood pressure with TCM therapy， it was found that the sample size was small and the preciseness needed to be improved. Further research is needed due to the complicated mechanism of TCM and acupuncture. There is a lack of insufficient evidence to support the long-term efficacy and safety of TCM therapy because of the limitations in clinical study intervention and follow-up time. This article reviews the clinical research of the effect of TCM on high-normal blood pressure， in order to provide guidance and reference for clinical and subsequent research.
Keywords：high-normal blood pressure;traditional Chinese medicine;review
Abstract：Diabetes is a kind of disease which poses a great threat to human health. Its occurrence and development are often related to many factors such as heredity and environment. According to the eighth edition of Diabetes Federation's diabetes map in 2017， there are about 425 million diabetics in the world. It is estimated that by 2045， the number of diabetics will increase to 700 million， becoming a health problem that has attracted increasing attention all over the world， among which the number of type 2 diabetics （T2DM）accounts for more than 90% of the total. Therefore， it is of great significance to study the pathological mechanism for the effective prevention and treatment of diabetes. Intestinal microflora coexists with human beings and forms an important micro ecosystem， which is involved in the metabolism of substance and energy. In recent years， with the development of high-throughput sequencing technology， a large number of studies have shown that in addition to obesity， genetic and insulin dysfunction， intestinal flora disorder may also lead to diabetes. The unbalanced diet structure of T2DM patients destroys the balance of intestinal flora. It is generally believed that the occurrence and development of T2DM may be one of the results of the intestinal microbial disorder caused by over nutrition. However， there is no clear mechanism of how intestinal flora participates in the development of T2DM. At present， it is generally believed that the intestinal flora may affect the metabolism of the body through the participation in bile acid metabolism， short chain fatty acid metabolism， low-level inflammatory response and other ways. At present， the prevention and treatment of T2DM is mainly based on drug control. Through surgical operation， increasing the number of probiotics， fecal transplantation and other methods to intervene the intestinal microflora to adjust the intestinal microflora， it provides a new means for the prevention and treatment of T2DM. This paper discusses the interaction between T2DM and intestinal microflora in recent years and the possible treatment measures in the future.