Abstract：Depression is a kind of emotional and mental disorder， which is related to many pathogenic factors， such as abnormal metabolism of monoamine neurotransmitters， immune inflammatory reaction， neuroendocrine disorder and so on. it has the characteristics of high disability rate and high recurrence rate， which is seriously harmful to human health. It brings huge economic burden to patients and their families. At present， the commonly used antidepressants in clinic are monoamine oxidase inhibitors， tricyclic antidepressants and selective 5-hydroxytryptamine reuptake inhibitors. Long-term use causes adverse reactions such as cardiovascular， urinary and digestive dysfunction. The multi-target， multi-pathway and multi-mechanism of traditional Chinese medicine has played a great role in the treatment and rehabilitation of depression. At the same time， traditional Chinese medicine has gradually become a hot spot in the research and development of antidepressants because of its small side effects and good drug compliance. Especially the ZHANG Zhong-jing's classical formulae with precise medication and rigorous prescription has unique advantages in the treatment of depression. For thousands of years， it has been used clinically by countless doctors in the treatment of various types of depression， with definite curative effects and few side effects. In recent years， clinical research， experimental research， and pharmacological research around ZHANG Zhong-jing's classical formulae for depression have been continuously expanded and deepened. This article reveals the rules of medication of the treatment of depression with ZHANG Zhong-jing's classical formulae from three aspects of Bupleurum class prescription， Gardenia class prescription， and other prescriptions， which is reflected in the study on the composition of prescription， the etiology， and pathogenesis and the symptoms of the treatment. Integrating and classifying ZHANG Zhong-jing's classical formulae's experience in the treatment of depression， mainly expounded from two aspects， clinical research and pharmacological research， summarizing the research perspective and progress of Zhongjing prescription medicine in the treatment of depression in recent years， to excavate， inherit and develop ZHANG Zhong-jing's classical formulae， and provide experience and reference for the treatment of depression from multiple angles and ideas.
Keywords：ZHANG Zhong-jing's classical formulae;depression;research progress;Bupleuri Radix class prescription;Gardeniae Fructus class prescription
Abstract：ObjectiveTo study the effect of Chaihu Jia Longgu Mulitang on phosphatidylinositol-3-kinases （PI3K）/protein kinase B （Akt）/glycogen synthase kinase 3β （GSK3β）/β-catenin signaling pathway of hippocampus in rats with depression.MethodA total of 120 SD rats were randomly divided into normal group，model group， and low， middle and high-dose Chaihu Jia Longgu Mulitang groups（3.25，6.5，13 g·kg-1）， and fluoxetine group， with 20 rats in each group. Except normal group， the depression model was prepared through chronic unpredictable mild stimulation（CUMS）. The normal group and the model group were given normal saline with 6.5 g·kg-1 by gavage. Chaihu Jia Longgu Mulitang groups were intragastrically given corresponding herbal drugs 3.75，6.5，13 g·kg-1， while fluoxetine group was intragastrically given fluoxetine 10 mg·kg-1 for 21 days， once a day. Then the depressive behaviors of rats were observed by sucrose preference test （SPT） and forced swimming test （FST）. Western blot was used to detect the protein expressions of PI3K，Akt，GSK3β and phosphorylation level.ResultCompared with normal group，the sucrose preference index was decreased significantly，while the immobility time in FST was increased significantly（P<0.01）， the protein expressions of PI3K， p-PI3K p110， p-PI3K p85 were decreased significantly （P<0.01）， and expressions of Akt， p-Akt Thr308，p-Akt Ser473， p-GSK3β Ser9 and β-catenin were decreased significantly（P<0.01）， while the level of GSK3β， p-GSK3β Tyr216 were increased significantly in model group（P<0.05，P<0.01）. Compared with model group，Chaihu Jia Longgu Mulitang could increase sucrose preference index and decrease the immobility time in FST（P<0.01）， the protein expressions of PI3K p110 and PI3K p85 was increased significantly （P<0.01）， levels of Akt Thr308，Akt Ser473， p-GSK3β Ser9， β-catenin were increased significantly （P<0.01）， whereas levels of GSK3β， and GSK3β Tyr216 were decreased significantly.ConclusionChaihu Jia Longgu Mulitang could increase protein expression and activity of PI3K in rat hippocampus， activate Akt， inhibit GSK3β kinase activity and prevent β-catenin from degradation， so as to increase PI3K/Akt pathway activity in rat hippocampus， and protect hippocampal neurons.
Abstract：ObjectiveTo study the effect of modified Suanzaoren Tang on the expression of excitatory amino acids receptor（EAARs） in hippocampus of rats with chronic depression， and to explore the anti-depressant mechanism of modified Suanzaoren Tang based on excitatory amino acids receptor.MethodSixty SD rats were randomly divided into normal group，model group，and low，middle and high-dose modified Suanzaoren Tang groups，and ketamine group，with 10 rats in each group．Except normal group，the depression model of rats was prepared by using chronic restraint stress（CRS）.The normal group and model group were intragastrically（ig） given normal saline．the modified Suanzaoren Tang groups were intragastrically given corresponding herbal drugs 6，12，24 g·kg-1， ketamine group group were given ketamine 0.015 g·kg-1 through intraoeritoneal injection，for 21 days，once a day．Then the depressive behaviors of rats were observed by Morris water maze and novelty feeding experiment.Western blot was used to detect the levels of DAR1，NMDAR2A，NMDAR2B，GluR1，mGluR1，CaMKⅡα and CaMKⅡβ protein expression in rat hippocampus tissue.ResultCompared with normal group，the time of novel ingestion and escape latencywere prolonged significantly（P<0.01）， and the time of space exploration was shortened significantly（P<0.01）.The levels of NMDAR1，NMDAR2A，NMDAR2B，mGluR1 and CaMKⅡβ expression were increased significantly（P<0.01），while the levels of GluR1 and CaMKⅡα expression were decreased significantly（P<0.01）in model group. Compared with model group，the time of novel ingestion and escape latency were shortened significantly （P<0.01）， and the time of space exploration was prolonged significantly（P<0.01）.The levels of NMDAR1，NMDAR2A，NMDAR2B，mGluR1 and CaMKⅡβ protein expression were decreased significantly（P<0.01），but the levels of GluR1 and CaMKⅡα expression were increased decreased significantly（P<0.01）in middle and high-dose modified Suanzaoren Tang groups.ConclusionModified Suanzaoren Tang can improve the behavior of chronic depression rats effectly. Its mechanism may be related with reduction the expression of NMDAR1，NMDAR2A，NMDAR2B，mGluR1 and CaMKⅡβ protein ，increase the expression of GluR1and CaMKⅡα protein.
Abstract：ObjectiveTo study the effect of modified Xiao Chaihutang on the expressions of excitatory amino acid transporters（EAATs） and vesicle glutamate transporters（VGLUTs）in hippocampus of rats with chronic depression， in order to explore the anti-depressant mechanism of modified Xiao Chaihutang based on glutamate transport.MethodA total of 120 SD rats were randomly divided into normal group， model group， and low， middle and high-dose modified Xiaochaihutang groups （6.5， 13， 26 g·kg-1） and riluzole group， with 20 rats in each group．Except normal group， the depression model of rats was prepared through Chronic restraint stress（CRS）. The normal group and the model group were intragastrically （ig） given normal saline. The modified Xiao Chaihutang groups were intragastrically given corresponding herbal drugs （6.5， 13， 26 g·kg-1）， and the Riluzole group was given Riluzole 20 mg·kg-1 through intraoeritoneal injection for 21 days， once a day. Then the depressive behaviors of rats were observed by forced swimming test （FST） and tail suspension test （TST）. The level of glutamic acid （Glu） in rats hippocampus was determined by high performance liquid chromatography （HPLC）. The mRNA expressions of EAAT1， EAAT2 and EAAT3 in hippocampus were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）method. Western blot was used to detect the protein expressions of EAAT1， EAAT2， EAAT3， VGLUT1 and VGLUT2 in rat hippocampus tissue. Nissl staining was used to observe the morphology of hippocampal neurons in rats. Immunohistochemical（IHC）S-P method were used to detect the location expressions of EAAT1， EAAT2 and NeuN proteins in rat hippocampal CA1 region tissue.ResultThe immobility times in FST and TST were increased significantly（P<0.01）， the mRNA and protein expressions of EAAT1，EAAT2，EAAT3 were decreased significantly （P<0.01）， and as well as the expressions of VGLUT1 and NeuN were decreased significantly（P<0.01）， while the level of Glutamate and the expression of VGLUT2 were increased significantly（P<0.01） in model group， compared with normal group. Compared with model group，the immobility times in FST and TST were decreased significantly（P<0.05， P<0.01）， mRNA and protein expressions of EAAT1，EAAT2，EAAT3 were increased significantly（P<0.01）， and expressions of VGLUT1 and NeuN were increased significantly（P<0.01）. However， the level of Glutamate and the expression of VGLUT2 were decreased significantly（P<0.01）， and the damage of hippocampal neurons in rats was mild in middle and high-dose modified Xiao Chaihutang groups.ConclusionModified Xiao Chaihutang has an anti-depressive effect. Its mechanism may be related to its up-regulation of expressions of EAAT1， EAAT2， EAAT3 genes and VGLUT1 protein in the hippocampus of depression model rats.
Abstract：Depression is a common emotional disease，causing a heavy burden of disease on patients' families and society.Its pathogenesis is not yet fully understood，andmay be related to inflammation，neurotransmitters，neurotrophicfactors，hippocampal neuronal synaptic plasticity，oxidative stress，etc.Clinical application of Western medicine to treat depression has serious side effects，poor patient compliance，the effective rate is not high，rebound after withdrawal，etc.Traditional Chinese medicine treatment of depression has the characteristics of low side effects and high patient compliance. Depression belongs to the category of depression syndromein traditional Chinese medicine.Traditional Chinese medicine believes that the main symptoms of depression include depressed liver Qi，stagnated Qi transforming into fire，deficiency in heart and spleen，deficiency in heart and lung，stagnation of phlegm and Qi，deficiency of liver and kidney，etc. Banxia Houpu Tang is a classic recipe contained in"Synopsis of the Golden Chamber" by Zhang Zhongjing in the Han Dynasty， it has the effect of treating depression syndrome of stagnation of phlegm and Qi.By searching the literatures of Banxia Houpu Tang in the treatment of depression in Chinese knowledge network database （CNKI），Chinese biomedical literature database （CBM）， Wanfang Data， Pubmed， Embase， Web of Science and other databases，we found that both clinical application and experimental research suggest that Banxia Houpu Tang has a significant antidepressant effect. Clinical studies have shown that Banxia Houpu Tang can improve the scores of anxiety scales and depression scales of patients with depression. The antidepressant effect is significant， and the advantages are prominent. Experimental research shows that the antidepressant effect of Banxia Houpu Tang and its effective components may be related to the factors such as intervention of inflammatory response， influence of neurotransmitters， regulation of neurotrophic factors， improvement of synaptic plasticity of hippocampal neurons and reduce oxidative stress，etc. Therefore，this paper reviews several types of depression in the clinical treatment of Banxia Houpu Tang and the related experimental studies of Banxia Houpu Tang.
Abstract：The prescription research and clinical application of Wenjingtang were summarized in order to provide reference for the formulation of material standard and the development of compound preparation. By systematically sorting out the relevant ancient medical books and modern literature reports， combined with the relevant policy requirements of the development of compound preparations of famous classical formulas， this paper expounded the existing problems and put forward some suggestions. Wenjingtang is composed of nine herbs， which is derived from Complete Effective Prescriptions for Women's Diseases written by CHEN Zi-ming in Song dynasty. The original medicinal plants and medicinal parts of the formula were basically the same as those recorded in the 2020 edition of Chinese Pharmacopoeia， for example， Paeonia lactiflora was the choice of Paeoniae Radix Alba， Cinnamomi Cortex was selected as Guixin， the rhizoma of Curcuma phaeocaulis was selected as Curcumae Rhizoma. It was suggested that raw products should be selected for decoction pieces and processed according to the methods recorded in the 2020 edition of Chinese Pharmacopoeia. According to the dosage of 1 Liang=40 g， the dosages of Angelicae Sinensis Radix， Chuanxiong Rhizoma， Paeoniae Radix Alba， Cinnamomi Cortex， Moutan Cortex and Curcumae Rhizoma were 20 g， the dosages of Ginseng Radix et Rhizoma， Glycyrrhizae Radix et Rhizoma and Achyranthis Bidentatae Radix were 40 g， the total amount of this formula was 240 g. The 9 kinds of decoction pieces were crushed （particle size<6 mm） separately and mixed， each dose was 20 g， 525 mL of water was added， and then decocted to 280 mL. After filtration， warm medicine was taken for once a day. According to ancient books， Wenjingtang has the functions of activating blood circulation， regulating menstruation，warming meridians and dispersing cold. It is mainly used to treat dysmenorrhea， irregular menstruation and other diseases in modern clinical practice. In addition， it has certain curative effect on endometriosis， pelvic inflammatory disease， acne， eczema and other skin diseases. Through the research， the historical evolution and clinical application of Wenjingtang are fully clarified， which can provide research for the later development and application of this famous classical formula.
Abstract：ObjectiveBy studying the effects of Tianwang Buxindan on sleep quality， cognitive function， inflammatory factors and immune-related gene expression in sleep deprivation model rats， explore the effect of Tianwang Buxindan on the learning and memory process under sleep deprivation and its anti-inflammatory effects possible mechanism.MethodThe 40 male SPF rats were used to simulate the sleep deprivation model by multi-platform water environment method， and were randomly divided into model group， Tianwang Buxindan group （20 g·kg-1） and estazolam group （0.1 mg·kg-1）， set up a normal group， 10 in each group. A total of 4 weeks of sleep deprivation modeling was performed， and drug intervention was performed 2 weeks later. The model group and the blank group were given equal volumes of pure water. Electroencephalogram （EEG） evaluation of modeling and analysis of sleep structure and quality of rats， Morris water maze positioning navigation and space exploration experiment analysis of learning and memory ability of rats， application of enzyme-linked immunosorbent assay （ELISA） was used to detect the serum inflammatory factor interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， monocyte chemokine-1 （MCP-1） expression， Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA levels of EB virus inducible gene 3 （EBI3）， extracellular signal-regulated kinase 5 （ERK5）， and p21 activated protein kinase 4 （PAK4）.ResultThe sleep deprivation model was successfully built. Compared with blank group， the total sleep time， total duration of slow wave sleep and the duration of the first and second phases of slow wave sleep in the model group were significantly shortened （P<0.01）. The incubation period on the upper platform， the total swimming distance and the time to reach the original platform for the first time increased significantly， while the number of times to cross the platform and the target quadrant significantly decreased （P<0.05，P<0.01）. The expression levels of IL-1β，TNF-α and MCP-1 increased significantly （P<0.01）， the mRNA expression levels of EBI3， ERK5 and PAK4 in the hypothalamus of the model group decreased significantly （P<0.01）. Compared with the model group， the sleep quality of the rats in the Tianwang Buxindan group was significantly improved. The total sleep time， the total duration of slow wave sleep and the duration of the first phase of slow wave sleep were significantly increased （P<0.01）. The incubation period on the platform， the total swimming distance and the time to reach the original platform for the first time are shortened， the number of times to cross the platform and the target quadrant time are extended （P<0.05，P<0.01）， IL-1β， TNF-α， MCP-1 expression levels were significantly reduced （P<0.05，P<0.01）， mRNA expression levels of EBI3， ERK5 and PAK4 in rat hypothalamus were significantly increased （P<0.05，P<0.01）.ConclusionTianwang Buxindan can improve the sleep quality and learning and memory ability of sleep deprivation model rats， which may be related to the increase of the expression level of related inflammatory factors and its anti-inflammatory effect.
Abstract：ObjectiveTo observe the effect of Wenjingtang on the expression of hypoxia inducible factor-1α （HIF-1α） and ultrastructure of mitochondria in rats with endometriosis （EMs）， and to explore the mechanism of Wenjingtang in the treatment of EMs.MethodThe EMs model rat was established by autogenous intimal transplantation， and the volume of ectopic lesions was measured by small animal ultrasound imaging system. According to the volume of ectopic lesions， rats successfully modeled were randomly divided into model group， low， middle ang high dose Wenjingtang group （4.85， 9.7， 19.4 g·kg-1） and gestrinone group （0.25 mg·kg-1）， 10 in each group， another 10 rats were recruited in a sham operation group. After 6 weeks of drug treatment， the volume of ectopic lesions was measured by ultrasound imaging system and caliper， the morphology of ectopic endometrium was observed by hematoxylin-eosin （HE） staining， the levels of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α） and transforming growth factor-β1 （TGF-β1） in peritoneal fluid were measured by enzyme linked immunosorbent assay （ELISA）， the mRNA and protein expression of HIF-1α in eutopic or ectopic endometrial tissues were detected by Real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） and Western blot， and the ultrastructure of mitochondria in ectopic endometrium was observed by transmission electron microscope （TEM）.ResultCompared with sham operation group， ectopic lesions were found in model group， showing vesicular structure， and the typical endometrial tissue morphology was found in the lesions. The contents of IL-1β， TNF-α， TGF-β1 and the expression of HIF-1α mRNA and protein were significantly increased than those in the sham operation group （P<0.05，P<0.01）. TEM showed that the number of mitochondria in the cytoplasm of ectopic endometrium increased and the structure was intact. Compared with model group， the volume of ectopic focus was significantly decreased in all Wenjingtang groups （P<0.01）， and the results of ultrasonic examination were basically consistent with those of caliper measurement， HE staining showed that the columnar epithelial cells of ectopic endometrium were damaged or exfoliated and the interstitial cells were loose， the content of TNF-α in each dose group of Wenjingtang was significantly decreased， the content of IL-1β and TGF-β1 in the medium dose and high dose groups of Wenjingtang was significantly decreased （P<0.05，P<0.01）， the expression of HIF-1α mRNA and protein were significantly decreased in all Wenjingtang groups （P<0.05，P<0.01）. The mitochondria of ectopic endometrium were obviously swollen， the crest was broken or even disappeared， some of the mitochondria were vacuolar degeneration and the outer membrane was ruptured.ConclusionWenjingtang has a good therapeutic effect on experimental EMs in rats， and the mechanism is related to reducing the expression of HIF-1α， improving hypoxia in ectopic lesions and inducing mitochondrial damage in ectopic endometrium.
Keywords：Wenjingtang;endometriosis;hypoxia inducible factor-1α;ectopic lesions volume;small animal ultrasound imaging;ultrastructure of mitochondria
Abstract：ObjectiveTo observe the immune factors， coagulation and curative effect of modified Shoutaiwan with Si Junzitang combined with dydrogesterone tablets in advanced age patients with early threatened abortion， and to explore its mechanism of action.MethodThe 90 advanced age patients with threatened abortion and kidney deficiency and blood stasis syndrome differentiation in traditional Chinese medicine （TCM） were randomly divided into control group and observation group by random number table， with 45 cases in each group. Both groups took oral dydrogesterone tablets for luteal support. The control group additionally received natural vitamin E soft capsules by oral administration， while observation group received modified Shoutaiwan with Si Junzitang. The course of treatment was 10 days in both groups. The clinical efficacy， TCM syndrome score， immune factors and coagulation factors of the two groups were compared before and after treatment.ResultThere was no statistically significant difference in TCM symptom scores， immune factors， and coagulation factors between two groups before treatment. After treatment， the scores of TCM syndromes were reduced in both groups （P<0.05）， the proportion of helper T lymphocyte （Th）， Th/Ts ratio， D-dimer （D-D） level and fibrinogen （FIB） were reduced while prothrombin time （PT） and the ratio of suppressor T lymphocyte （Ts） were increased in observation group （P<0.05）. After treatment， the proportion of Th， Th/Ts， D-D， and FIB levels in observation group were lower than those in control group， while PT and the proportion of Ts were higher than those in control group （P<0.05）. The proportion of natural killer cells （NK） had no significant change after treatment， also with no significant difference between two groups. The total effective rate was 84.4%（38/45） in observation group， higher than 64.4%（29/45） in control group （χ2=4.398，P<0.05）. There was no obvious adverse reaction in both groups during the treatment.ConclusionModified Shoutaiwan with Si Junzitang combined with dydrogesterone tablets can improve symptoms and the therapeutic effect for fetal protection by regulating the immune balance and coagulation function in advanced age patients with threatened abortion.
Keywords：advanced age;threatened abortion;modified Shoutaiwan with Si Junzitang;didroxyprogesterone tablets;immune factor;coagulation factor
Abstract：ObjectiveTo observe the preventive and therapeutic effects of 5-week administration with Xiaoyaosan on rat liver injury caused by tripterygium Glycosides.MethodThirty-one SD rats were randomly divided into 4 groups， namely normal group， tripterygium glycosides group， tripterygium glycosides+Xiaoyaosan group （treatment group）， and tripterygium glycosides+Xiaoyaosan for 1 week in advance group （prevention group）. Tripterygium glycosides （37.5 mg·kg-1） was administered intragastrically， and Xiaoyaosan （water decoction， 19.270 g·kg-1） was administered intrastrically. First， the rats of prevention group were intragastrically administrated with Xiaoyaosan at 8：00-9：00 am， and the rats of other groups were given an equal volume of normal saline. After 1 week， the rats of tripterygium glycosides group were administered intragastrically with tripterygium glycosides suspension at 8：00-9：00 am. The rats of the treatment group and the prevention group were intragastrically administrated with Xiaoyaosan at 8：00-9：00 am， and then tripterygium glycosides suspension 2 hours later. All the drugs were given once a day for 5 weeks. Hematoxylin-eosin （HE） staining was used to observe the pathological changes of liver， enzyme-linked immunosorbent assay （ELISA） was used to detect serum interleukin-1β （IL-1β）， interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α）， chemical method was used to detect the content changes of aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， glutathione peroxidase （GSH-Px）， superoxide dismu-tase （SOD）， trace malondiamine aldehyde （MDA）. Immunohist-ochemical staining was used to observe the expressions of IL-6， IL-1β and TNF-α in liver tissue.ResultCompared with the normal group， the tripterygium glycosides group showed inflammatory cell infiltration， hepatocyte edema， hepatic sinuses squeezing and narrowing， liver plate widening， and liver cell necrosis， the contents of serum IL-1β， IL-6， TNF-α， ALT， AST and MDA were significantly increased（P<0.01）， but the contents of GSH-Px and SOD were decreased significantly （P<0.05， P<0.01）， while the positive expressions of IL-6， TNF-α， IL-1β in liver tissue were significantly increased （P<0.01）. Compared with tripterygium glycosides group， rats in the treatment group and the prevention group had less inflammatory cells infiltration and reduced edema in the liver tissue， and disorders in some cell， the contents of serum IL-1β， IL-6， TNF-α， ALT， AST， MDA were significantly decreased （P<0.01）， the contents of GSH-Px and SOD were significantly increased （P<0.05）， and the positive expressions of IL-6， TNF-α， IL-1β in liver tissues were significantly decreased （P<0.01）， with a better efficacy in the prevention group.ConclusionXiaoyaosan can obviously alleviate the long-term liver toxicity caused by tripterygium polyglycoside to a certain extent， with a better prophylactic effect.
Abstract：ObjectiveTo study the protective effect of different doses of single-flavored Coptis， Magnoliae Officinalis Cortex， and their compatibility on ulcerative colitis （UC） model rats and the colonic B lymphoblastoma-2 associated X protein （Bax） and cysteine-containing aspartame-3（Caspase-3） protein， inflammatory cytokines， and other expressions.MethodThe 120 healthy adult SD rats were randomly divided into blank group， model group， sulfasalazine group， Coptidis Rhizoma 2.00， 1.00， 0.50 g·kg-1 group， Magnoliae Officinalis Cortex 2.00， 1.00， 0.50 g·kg-1 group， Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00， 2.00， 1.00 g·kg-1 group， 12 groups with 10 rats in each group. The UC model was prepared by 2，4， 6-trinitrobenzene sulfonic acid/ethanol （TNBS/ethanol）. After 24 h of modeling， the rats were gavaged at 10 mL·kg-1 for one time/d. After modeling， the mental state， activity state， hair luster， stool characteristics， and blood in the stool of each group were observed. After continuous administration for 6 days， colon tissues and spleen were taken after the last administration for 24 h. The ratio of colonic weight to length and spleen index was calculated. The degree of colonic injury was evaluated according to the colonic mucosal injury index （CMDI） score criteria. the histopathological observation was performed using hematoxylin-eosin staining （HE）. The expression levels of tumor necrosis factor （TNF）-α， interleukin-6 （IL-6）， interleukin-10 （IL-10）， and myeloperoxidase （MPO） in the serum of Coptidis Rhizoma 2.00 g·kg-1 group， Magnoliae Officinalis Cortex 2.00 g·kg-1 group， Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00 g·kg-1 were detected by enzyme-linked immunosorbent assay（ELISA） in blank group and model group. Western blot was used to detect the expression of Bax and Caspase-3 proteins in the colon of rats.ResultCompared with blank group， rats in model group were sluggish and less active. The colon weight-length ratio， spleen index， CMDI， and colon tissue pathological damage increased significantly， and the expression of serum TNF-α， IL-6， and MPO increased significantly. Serum IL-10 expression levels were extremely significantly reduced （P<0.01）. Compared with model group， the sulfasalazine group， the Coptidis Rhizoma 2.00， 1.00 g·kg-1 group， the Magnoliae Officinalis Cortex 2.00 g·kg-1 group， and the three-dose groups of Coptidis Rhizoma combine with Magnoliae Officinalis Cortex， their colon weight-length ratio and CMDI were significantly reduced （P<0.05，P<0.01）. The colon weight length ratio and CMDI index of the Coptidis Rhizoma 0.50 g·kg-1 group， Magnoliae Officinalis Cortex 0.50 and 1.00 g·kg-1 group were not significantly different from the model group but compared with Coptidis Rhizoma and Magnolia 0.50 g·kg-1 group， the ratio of colon weight to length in the group of Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 1.00 g·kg-1 group was significantly reduced （P<0.01）. Compared with model group， the spleen index of the sulfasalazine group， the Coptidis Rhizoma 2.00 g·kg-1， and the Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00 g·kg-1 group were significantly lower （P<0.05）， compared with model group， the sulfasalazine group， Coptidis Rhizoma 2.00， 1.00 g·kg-1 and Magnoliae Officinalis Cortex 2.00 g·kg-1， thre dose groups of Coptidis Rhizoma combine with Magnoliae Officinalis Cortex can significantly improve the depth and scope of histopathological damage and tissue necrosis. Compared with the model group， the preferred Coptidis Rhizoma 2.00 g·kg-1 group， Magnoliae Officinalis Cortex 2.00 g·kg-1 group， Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00 g·kg-1 group serum TNF-α， IL-6， MPO expression levels are extremely significantly reduced， the level of IL-10 increased significantly （P<0.01）．Compared with blank group， the expression of Bax and Caspase-3 protein in the colon of model group was significantly increased （P<0.01）. Compared with model group， the expression of Bax and Caspase-3 protein in preferred Coptidis Rhizoma 2.00 g·kg-1 group and Magnoliae Officinalis Cortex 2.00 g·kg-1 group， Coptidis Rhizoma combine with Magnoliae Officinalis Cortex 4.00 g·kg-1 group were significantly reduced （P<0.01）．ConclusionThe compatibility of single-flavored Coptidis Rhizoma， Magnoliae Officinalis Cortex， and Coptidis Rhizoma combine with Magnoliae Officinalis Cortex may improve the pathology of UC model rats induced by TNBS/ethanol by down-regulating the expression of Bax and Caspase-3 protein， inhibiting the release of inflammatory cytokines and promoting the release of anti-inflammatory factors injury， it plays a role in protecting colonic mucosa. The compatibility effect of Coptidis Rhizoma and Magnoliae Officinalis Cortex is better than that of single medicine， and Coptidis Rhizoma has a tendency to be better than Magnoliae Officinalis Cortex.
Abstract：ObjectiveTo observe the effect of oral administration of Tianlong Tongxin tablet on acute myocardial ischemia and related indexes in experimental dogs.MethodThe model of acute myocardial ischemia in dogs was established and the dogs were divided into the control group （equal amount of normo-cyclodintrin 10 g·kg-1）， Hexinshuang group （5 mg·kg-1）， Tianlong Tongxin tablet high， medium and low dose groups （1， 0.5， 0.25 g·kg-1） and the compound Danshen tablet group （0.144 g·kg-1）. Myocardial ischemia degree was measured by epicardium electrocardiogram， the range of myocardial infarction was determined by quantitative histology （N-BT staining）， and coronary blood flow， cardiac output， myocardial oxygen consumption， coronary resistance and peripheral resistance were measured. Meanwhile， serum creatine kinase （CK）， lactate dehydrogenase （LDH） and serum superoxide dismutase （SOD） activity and malondialdehyde （MDA） content were detected by optical kit.ResultAs compared with the control group， Tianlong Tongxin tablet can reduce the myocardial ischemia degree （∑-ST） measured by the electrocardiogram of the pericardium （P<0.05）， reduce the infarcted area shown by N-BT staining （P<0.05）， reduce the venous oxygen content （P<0.05）， increase the coronary flow， cardiac output and myocardial oxygen consumption of anesthetized dogs， and reduce the coronary artery resistance and peripheral resistance （P<0.05）. However， there was no significant difference in the influence of serum CK， LDH， SOD activity and MDA content in serum.ConclusionTianlong Tongxin tablet can improve acute myocardial ischemia and myocardial infarction in dogs.
Abstract：ObjectiveTo investigate the inhibitory effect and the possible mechanism of essential oil from fructus Alpinia zerumbet （EOFAZ） on endothelial-to-mesenchymal transition （EndMT） induced by high glucose （HG）.MethodHuman umbilical vein endothelial cells （HUVECs） was cultured in vitro to analyze the pharmacodynamic effects of EOFAZ on EndMT and oxidative stress damage induced by HG. The experiment was set the blank group， HG group （35 mmol·L-1）， EOFAZ low dose group （1 μg·L-1） and EOFAZ high dose group （4 μg·L-1）. After EOFAZ intervention for 2 h， HG was added to incubate for 72 h in order to establish EndMT cell model. Western blot was used to detect the protein expression of vimentin and platelet endothelial cell adhesion molecule （CD31）. Angiogenesis experiment was used to detect the ability of cell migration ability in order to analyze the effect of EOFAZ on EndMT. The changes of reactive oxygen species （ROS） levels were detected by 2'，7'-dichlorodihydrofluorescein diacetate （DCFH-DA） fluorescence probe and the contents of malondialdehyde （MDA）， superoxide dismutase （SOD） and catalase （CAT） in cells were detected by the kit method to analyze the effect of EOFAZ on oxidative stress. Western blot was used to detect the protein expression levels of nuclear transcription factor E2 related factor 2 （Nrf2） and Notch1. The overexpression of Nrf2 was achieved by adenovirus （AD） transfection and the mechanism of EOFAZ inhibiting EndMT was further analyzed. The experiment was set the blank group， HG group （35 mmol·L-1）， AD-Nrf2 group， EOFAZ group （4 μg·L-1）， AD-Nrf2+EOFAZ group （4 μg·L-1）. The cells were infected with recombinant adenovirus overexpression plasmid of Nrf2 gene for 6 h， then replaced with normal medium for 24 h. After EOFAZ intervention for 2 h， HG was added to co-incubate for 72 h to induce EndMT. Western blot was used to detect the protein expressions of Nrf2， CD31， vimentin， Notch1 and Snail.ResultCompared with the HG group， after treatment with EOFAZ， the protein expression of CD31 was significantly up-regulated （P<0.05）， the protein expression of vimentin was significantly down-regulated （P<0.01）， the ability of cell migration was decreased （P<0.01）， and the contents of ROS and MDA were decreased （P<0.05， P<0.01）， the levels of CAT and SOD were increased （P<0.01）. In addition， EOFAZ could significantly up-regulate the protein expression of antioxidant signal Nrf2 （P<0.01） and down-regulate the protein expression of Notch1 （P<0.01）. High expression of Nrf2 was achieved by stable AD transfection into HUVECs. The results of Western blot showed that， compared with the HG group， the protein expression levels of Nrf2 and CD31 in each treatment group were significantly increased （P<0.01）， while the protein expression levels of vimentin， Notch1 and Snail were down-regulated （P<0.01）. At the same time， compared with the AD-Nrf2 group， the AD-Nrf2+EOFAZ group could further up-regulate the protein expressions of Nrf2 and CD31 （P<0.05， P<0.01）， while decrease the protein expression levels of vimentin， Notch1 and Snail （P<0.01）.ConclusionEOFAZ ameliorates oxidative stress injury of vascular endothelial cells induced by HG and inhibits EndMT， which is related to Nrf2/Notch1 signaling pathway.
Keywords：Alpinia zerumbet fructus;essential oil;oxidative stress;endothelial-to-mesenchymal transition （EndMT）;nuclear transcription factor E2 related factor 2 （Nrf2）;Notch signal;Western blot
Abstract：ObjectiveTo investigate the effect of Ru′ai Shuhou prescription （RSR） drug-containing serum on the proliferation and invasion ability of breast cancer cells MDA-MB-453 based on the biological axis of stromal cell-derived factor-1（SDF-1）/chemokine receptor 4 （CXCR4）.MethodA model of MDA-MB-453 cells with SDF-1-induced high expression of CXCR4 was established， and the rat drug-serum containing RSR and blank rat serum were prepared respectively. The cells were divided into fetal bovine serum control group （Blank）， blank rat serum group， SDF-1+blank rat serum group， SDF-1+RSR group， AMD3100+ SDF-1+blank rat serum group， and AMD3100+ SDF-1+RSR group. After intervention for 48 h， cell proliferation was detected by cell counting kit-8 （CCK-8） assay， cell invasion ability was detected by transwell assay， and mRNA and protein expressions of CXCR4， matrix metalloproteinase-2 （MMP-2） and MMP-9 were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively.ResultAs compared with the blank serum group， the proliferation of MDA-MB-453 cells was promoted and expression of CXCR4 mRNA was increased significantly when SDF-1 was 100 μg·L-1 （P<0.05）. As compared with SDF-1+blank rat serum group， RSR inhibited the proliferation and invasion of MDA-MB-453 cells induced by SDF-1， and at the same time， down-regulated the mRNA and protein expressions of CXCR4， MMP-2 and MMP-9 （P<0.05）. After pre-treatment with AMD3100 for 24 h， the inhibitory effect of RSR to cell proliferation was significantly increased （P<0.05）， and meanwhile， the decreases in mRNA and protein expression of CXCR4， MMP-2 and MMP-9 were more obvious， with statistically significant differences （P<0.05）.ConclusionThrough SDF-1/CXCR4 biological axis， RSR could down-regulate the expression of MMP-2 and MMP-9， reduce the degradation of extracellular matrix （ECM）， and then inhibit the metastasis of MDA-MB-453 cells. In addition， it has a synergistic effect with CXCR4 inhibitor AMD3100.
Abstract：ObjectiveTo study the anti-colon cancer effect and mechanism of magnolol analogue CT2-3， in order to lay a foundation for the application of CT2-3 in anti-colon cancer area.MethodColon cancer cells SW480 and LoVo were cultured in vitro. Different concentrations （10， 20， 40， 80 μmol·L-1） of CT2-3 and magnolol were used to stimulate colon cancer cells for 24， 48 h. The effect of CT2-3 and magnolol on the cell viability of colon cancer cells was detected by cell counting kit （CCK-8）. Colony formation assay was used to detect the colony formation capacity of CT2-3 on colon cancer cells. Flow cytometry and Western blot were used to determine the effect of CT2-3 on the apoptosis of colon cancer cells and the expression of DNA damage marker phosphorylated histone H2AX （γH2AX）. Reactive oxygen species （ROS） generation was measured by ROS assay kit. Real time quantitative polymerase chain reaction （Real-time PCR） was used to detect the effect of CT2-3 on expressions of mitochondrial apoptosis-related genes B-cell lymphoma-2 （Bcl-2） and Bcl-2-associated X （Bax） in colon cancer cells.ResultThe half maximal inhibitory concentration （IC50） of magnolol in two kinds of colon cancer cells after treatment for 24， 48 h were both higher than 80 μmol·L-1. While the IC50 of CT2-3 in SW480 cells after treatment for 24， 48 h were （54.59±1.73） μmol·L-1 and （29.82±1.13） μmol·L-1， respectively. The IC50 of CT2-3 in LoVo cells after treatment for 24，48 h were （66.68±2.11） μmol·L-1 and （46.70±1.81） μmol·L-1， respectively. Compared with the blank group， the colony formation capacity of colon cancer cells in CT2-3 groups （20， 40 μmol·L-1） was significantly decreased in a dose-dependent manner （P<0.01）， apoptotic colon cancer cells were significantly increased （P<0.01）， relative expression of DNA damage marker γH2AX was significantly increased （P<0.01）， ROS was significantly increased （P<0.01）. In addition， relative mRNA expression of Bcl-2 was significantly decreased （P<0.01）， while relative mRNA expression of Bax was significantly increased （P<0.01）.ConclusionCT2-3 can remarkably inhibit colon cancer cells， and the underlying mechanism might be that CT2-3 promotes mitochondria dysfunction and ROS generation by regulating expressions of mitochondrial apoptosis-related genes， so as to further induce DNA damage and finally lead to apoptosis.
Keywords：magnolol;magnolol analogue CT2-3;colon cancer;apoptosis;reactive oxygen species （ROS）
Abstract：ObjectiveTo observe the efficacy and safety of Qifu Huazhuo decoction in treating intermittent and chronic gout.MethodA total of 160 eligible patients with intermittent and chronic gout were divided into intervention group and control group according to treatment methods， 80 cases in each group. Qifu Huazhuo decoction combined with febuxostat was given in intervention group by oral administration， while the patients in control took febuxostat combined with sodium bicarbonate tablet. After twelve weeks of treatment in both groups， the onset frequency of gout， the degree of joint pain， the levels of uric acid， creatinine， alanine transaminase in the blood， number of cases to use analgesics and adverse events were recorded compared before and after treatment. Twelve weeks follow-up was also conducted after the treatment.ResultThe total effective rate was 91.25% （73/80） in intervention group， higher than 83.75% （67/80） in control group， with a significant difference between two groups （χ2=0.16， P<0.05）. Intervention group was superior to control group in improvement of onset frequency of gout， joint pain （P<0.05）， and blood uric acid （P<0.05）. There was no significant difference between two groups in blood creatinine and alanine transaminase. The number of patients using analgesics was less in intervention group （P<0.05）. In terms of adverse reactions， there were two cases of fever， two cases of abnormal liver function， six cases of abnormal kidney function and three cases of gastrointestinal reaction after the treatment in intervention group. While for control group， there were one case of fever， two cases of abnormal liver function， eight cases of abnormal kidney function and five cases of gastrointestinal reaction， without significant differences between two groups.ConclusionThe Qifu Huazhuo decoction can effectively reduce the onset frequency of gout and the degree of joint pain in intermittent and chronic phases， effectively control the level of blood uric acid， and reduce the number of cases using analgesics， with less adverse reactions， so it is suitable for clinical use.
Abstract：ObjectiveTo discuss the efficacy and safety of Shenwu Yishenpian on stage 4-5 chronic kidney disease-nondialysis （CKD） with deficiency of spleen and kidney Qi， and the effect on renal interstitial fibrosis （RIF） and microinflammation.MethodOne hundred and twenty patients were randomly divided into observation group and control group. A total of 58 patients in control group completed the treatment （including 2 patients falling off or lost）. And 58 patients in observation group completed the treatment （including 1 patient was falling off or lost visit， and 1 eliminated）. Both groups got comprehensive treatment of western medicine. Patients in control group got simulated medicine of Shenwu Yishenpian， 4 pieces/time， 3 times/day. Patients in observation group got Shenwu Yishenpian， 4 pieces/time， 3 times/day. The treatment lasted for 6 months until the renal replacement therapy， and the 6-month follow-up was recorded. For every month， blood creatinine （SCr） was detected， and glomerular filtration rate （eGFR） were calculated. The 12-month renal replacement （dialysis or kidney transplantation）， progress （CKD4 to CKD5） and mitigation （CKD5 to CKD4 or CKD4 to CKD3） were recorded. Before and after treatment， levels of urea nitrogen （BUN）， hemoglobin （HB）， plasma albumin （ALB）， urine protein quantity （24 hUp） and blood uric acid （UA） levels were detected， deficiency of spleen kidney Qi was scored， and transforming growth factor-β1 （TGF-β1）， connective tissue growth factor （CTGF）， serum Klotho， tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， interleukin-1 （Lkn-1） and interleukin-12 （IL-12） were detected. And the safety was evaluated.ResultAt the 3th and 6th after treatment， SCr in two groups increased （P<0.01）， while eGFR decreased （P<0.01）. Compared with control group， SCr was less than that in control group （P<0.01）， whereas eGFR was higher than that in control group （P<0.01）. During 12 months of observation， the reduction rate of CKD was 13.79% （8/58）， which was higher than 1.72% （1/58） in control group. The progress rate of CKD was 11.43% （4/35）， which was lower than 31.58% （12/38） in control group （P<0.05）. Levels of BUN， 24 hUp and UA were lower than those in control group （P<0.01）， while levels of Hb and ALB were higher than those in control group （P<0.01）. Effect in observation group was better than that in control group （Z=2.051， P<0.05）. And levels of TGF-β1， CTGF， TNF-α， IL-6， Lkn-1 and IL-12 were lower than those in control group （P<0.01）， and level of Klotho was higher than that in control group （P<0.01）. There was no adverse reaction relating to Shenwu Yishen Pian.ConclusionShenwu Yishenpian can delay the progress of renal function and CKD， reverse the progress of renal function in some patients， reduce the risk factors of disease progress， reduce the state of micro inflammation and resist RIF， and protect or improve renal function. Its clinical effect is better than placebo， and it is safe to use.
Keywords：chronic kidney disease;deficiency of spleen and kidney Qi;Shenwu Yishenpian;renal interstitial fibrosis;micro inflammatory state;prognosis of disease
Abstract：ObjectiveTo evaluate effect of addition and subtraction therapy of Buyang Huanwutang combined with Si Junzitang and acupuncture to poststroke fatigue （PSF） and syndrome of Qi deficiency and blood stasis， at the same time we studied the antioxidant and anti-inflammatory effects.MethodOne hundred and forty-four patients were randomly divided into control group and observation group （1∶1） by random number table. 66 patients in control group completed the therapy （4 patients were falling off or missing visit， 2 patients were eliminate）， 67 patients in observation group completed the therapy （2 patients were falling off or missing visit， 3 patients were eliminate）. In control group， patinets got acupuncture， 1 time/day， 6 times/week， they also got Geqi Tongmai grain， 10 g/time， 3 times/day. Patients in observation group got acupuncture （the same as which in control group）， and addition and subtraction therapy of Buyang Huanwutang combined with Si Junzitang， 1 dose/day， and courses of treatment in two groups were 4 weeks. Before and after treatment， fatigue severity scale （FSS）， NIH stroke scale （NIHSS）， syndrome of Qi deficiency and blood stasis， stroke specific quality of life scale （SS-QOL）， and scores of ability of daily life （ADL） were recorded. And levels of superoxide dismutase （SOD）， malondialdehyde （MDA）， glutathione peroxidase （GSH-Px）， homocysteine （Hcy）， interleukin-1β （IL-1β）， IL-6， tumor necrosis factor-α （TNF-α） and C-reactive protein （CRP） were detected. And the safety evaluated.ResultDisease effect in observation group was better than which in control group （Z=2.118， P<0.05）. And effect after using traditional Chinese medicine （TCM） was also better than that in control group （Z=2.046， P<0.05）. Scores of FSS， syndrome of Qi deficiency and blood stasis， NIHSS， and levels of IL-1β， IL-6， Hcy， CRP， TNF-α and MDA in observation group were all lower than those in control group （P<0.01）， and scores of SS-QOL， ADL， and levels of GSH-Px and SOD were all higher than those in control group （P<0.01）. Then there was no related safety issues caused by drug.ConclusionAddition and subtraction therapy of Buyang Huanwutang combined with Si Junzitang and acupuncture had effect of anti-oxidation and anti-inflammatory， and can significantly reduce fatigue and degree of neurological impairment and can improve patients' quality of life and daily life ability. The clinical effect is significant and safe， which is worthy of further research and application.
Keywords：poststroke fatigue;syndrome of Qi deficiency and blood stasis;Buyang Huanwutang;Si Junzitang;acupuncture;antioxidant;anti-inflammatory
Abstract：ObjectiveTo quantitatively analyze the changes of Staphylococcus aureus in different processed products of Angelicae Sinensis Radix.MethodThe real-time fluorescence quantitative polymerase chain reaction method （Real-time PCR） was established to quantitatively analyze S. aureus in Angelicae Sinensis Radix decoction pieces which bought from different producing areas， different enterprises and different storage time. The fluorescence quantitative reaction system was SYBR Premix Ex Taq Ⅱ of 10 μL， each of forward primer and reverse primer （10 μmol·L-1） of 0.8 μL， template/genome DNA of 1 μL， double distilled water of 7.4 μL. The reaction conditions of the fluorescence quantitative amplification curve were pre-denaturing for 30 s at 94 ℃， denaturing for 10 s at 94 ℃， annealing for 12 s at 60 ℃， extensing for 30 s at 72 ℃， cycling 45 times， single-point detection signal at 72 ℃. The melting curve was made from 72 ℃， and the step temperature of 0.5 ℃ was kept for 15 s to collect fluorescence. According to the results of Real-time PCR， representative samples were selected from Angelicae Sinensis Radix decoction pieces for comparison between plate counting method and Real-time PCR.ResultThe content of S. aureus in different processed products was sorted by rank of raw Angelicae Sinensis Radix>soil-fried Angelicae Sinensis Radix>wine-processed Angelicae Sinensis Radix. The content of S. aureus was the lowest in the samples from Weiyuan area of Gansu province by comparing with other producing areas. Compared with the retail enterprises， the content of S. aureus in raw products and wine-processed products from production and sale enterprises was lower. Different storage time had certain effect on the content of S. aureus in raw products and wine-processed products， and the content of S. aureus increased with the increase of storage time. The detection results of plate counting method were 3-4 orders of magnitude lower than that of Real-time PCR.ConclusionThe established Real-time PCR is superior to plate counting method in specificity， sensitivity， reliability and reporting period， which can provide an effective method for rapid and accurate quantitative detection of S. aureus in different processed products of Angelicae Sinensis Radix.
Abstract：ObjectiveTaking ultrafine granular powder of Salviae Miltiorrhizae Radix et Rhizoma （UGPSMR） as the research object， to establish a method for evaluating its physical properties.MethodA method was established for measuring the particle size distribution and specific surface area of UGPSMR， and the methodological investigation was carried out. A total of 15 physical indicators ［D90 （particle size value when the cumulative particle distribution reaches 90%）， particle size distribution range， particle size distribution width， bulk density， tap density， intergranular porosity， Carr index， specific surface area， pore volume， angle of repose， tablet angle， Hausner ratio， black to white degree L*， red to green degree a*， yellow to blue degree b*］ were used to characterize the quality attributes of UGPSMR and to construct the physical fingerprint. Multivariate statistical analysis methods such as similarity analysis， cluster analysis （CA）， principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA） were used to evaluate the quality of 11 batches of UGPSMR （S1-S11） produced in different years， and to find out the difference index between samples from different batches.ResultThe method for measuring the particle size distribution and specific surface area of UGPSMR was feasible and repeatable. The similarities between the physical fingerprint of 10 batches of samples （S1-S3， S5-S11） from production and control fingerprint of UGPSMR were above 0.85， but the similarity between sample S4 and the control fingerprint was only 0.488. There were some differences in physical property indicators between different batches of UGPSMR， and the characteristic difference indicators were intergranular porosity， specific surface area， pore volume， b*， L*， Carr index， particle size distribution width， respectively.ConclusionThis method can comprehensively evaluate the physical quality attributes of UGPSMR， and can reflect the effect of differences in material basis of the medicinal materials or production process on the physical properties of finished products， and can evaluate quality consistency between batches from the physical state level， which provides new ideas for the quality control of ultrafine granular powder of herbal medicine.
Keywords：ultrafine granular powder of Salviae Miltiorrhizae Radix et Rhizoma;physical property;particle size;comprehensive quality evaluation;physical fingerprint;multivariate statistical analysis
Abstract：ObjectiveTo study the volatile oil of Notopterygium inchum and N. franchetii by using gas chromatography-mass spectrometry （GC-MS）.MethodTen batches of N. inchum and N. franchetii were harvested from Sichuan and Gansu，including eight batches of wild samples， one batch of cultivated N. inchum and one batch of cultivated N. franchetii. Volatile oil was extracted according to the methods in Chinese Pharmacopeia， and the chemical constituents were analyzed with GC-MS technology.ResultNinety-seven chemical compounds were identified，including monoterpenes，sesquiterpenes and non-terpenes. The calculation results of the relative content of chemical components showed that α-pinene（25.39%-41.68%），β-pinene（19.34%-40.95%） and limonene（4.59%-11.99%） were main compounds in volatile oil of N. inchum and N. franchetii. Relative content of α-pinene was higher than that of β-pinene in wild N. inchum and cultivated N. franchetii， while relative content of β-pinene was higher than that of α-pinene in wild N. franchetii. Relative content of β-pinene in samples stored for one year was lower than that of newly harvested ones. The partial least squares discriminant analysis with SIMCA-P+11.5 software showed that the samples from Gansu and Sichuan provinces had no significant differences， and additionally，one-year-stored samples and newly-harvested samples were clustered into two groups.ConclusionThis study showed that the storage had influence on the chemical constituents of volatile oil in N. inchum， and it also revealed the differences in volatile oil components of wild and cultivated Notopterygium. The above mentioned work provided chemical basis for the pharmacology comparison study of volatile oil from different kinds of N. inchum and N. franchetii.
Abstract：ObjectiveTo investigate the allelopathic effects of water extracts from rhizosphere soil of three medicinal plants Rehmannia glutinosa，Pinellia ternata and Isatis indigotica on seed germination and seedling growth of Polygala tenuifolia， screen the stubble varieties suitable for crop rotation with P. tenuifolia， and provide some scientific basis for continuous cropping obstacles of P. tenuifolia.MethodThe bioassay method was used to study the effects of rhizosphere soil water extracts from three medicinal plants Rehmannia glutinosa，Pinellia ternata and Isatis indigotica at concentrations of 0.3，0.6，0.9 g·mL-1 on the germination of P. tenuifolia seed and seedling growth.ResultThe rhizosphere soil water extracts of Rehmannia glutinosa and Pinellia ternata showed basically low-promotion and high-inhibition concentration effects on the final germination rate，germination potential，and germination index of P. tenuifolia seeds，while the water extract of Isatis indigotica showed significant allelopathic inhibition effect. All three rhizosphere soil water extracts showed significant allelopathic inhibition effects on the growth index of P. tenuifolia seedlings. Among them，the rhizosphere soil water extract of Rehmannia glutinosa showed lower inhibitory effect on the plant height and root length of P. tenuifolia seedlings than the other two water extracts. The photosynthetic pigment content，proline（Pro） content，and soluble sugar content of P. tenuifolia chinensis seedlings were the highest under 0.3 g·mL-1 soil water extract of Rehmannia glutinosa， with relatively higher content of soluble protein， and relatively lower content of hydrogen oxide（H2O2）. Under the treatment of 0.9 g·mL-1 soil water extract of Rehmannia glutinosa，P. tenuifolia seedlings had the highest peroxidase（POD） and superoxide dismutase（SOD） activities，low catalase（CAT） activity，and lowest content of malondialdehyde（MDA）.ConclusionBased on the comprehensive analysis of the above experimental data and allelopathic effects，the water extract of rhizosphere of Rehmannia glutinosa can promote the germination of P. tenuifolia seeds to a certain extent，and lay the foundation for seedling resistance to biochemical stress. Therefore， Rehmannia glutinosa is more suitable for crop rotation with P. tenuifolia.
Keywords：Polygala tenuifolia;seed germination;rhizosphere soil water extract;allelopathic effect
Abstract：ObjectiveTo investigate constituents containing phenolic hydroxyl or carboxylic acid （excluding diarylheptanoids） from Curcumae Rhizoma and Sparganii Rhizoma herbal pair and single herb.MethodMultiple chromatographic separation techniques， including silica gel，MCI gel and Sephadex LH-20 gel， were employed to isolate and purify the compounds. Their structures were identified by means of the nuclear magnetic resonance （NMR），mass spectrometry （MS） and physicochemical properties. Constituents were quickly analyzed by UPLC-LTQ-Orbitrap-MSn，and scanned in positive and negative ion modes.ResultThese compounds were determined as trans-p-hydroxycinnamic acid（1），vanillic acid（2），protocatechuic acid（3），fumalic acid（4），succinic acid（5），succinic acid monomethyl ester（6），docosanoic acid（7），azelaic acid（8） and p-hydroxybenzaldehyde（9）. Forty compounds were speculated by comparing mass spectrometry data，retention times of some compounds and reference materials，including 14 phenols and 26 organic acids. Among the compounds of herbal pair，8 phenols and 22 organic acids in Sparganii Rhizoma as well as 11 phenols and 13 organic acids in Curcumae Rhizoma were identified. Cleavage pathways of main compounds were described.ConclusionThere are abundant phenols and organic acids in Curcumae Rhizoma and Sparganii Rhizoma herbal pair and single herb. The results enrich pharmacodynamic material basis of Curcumae Rhizoma and Sparganii Rhizoma herbal pair.
Abstract：Atherosclerosis （AS） is a chronic inflammatory disease which has great threat to human health in our society. The coronary heart disease， stroke and other cardiovascular and cerebrovascular diseases caused by it have a high mortality and disability rate. AS is characterized by endothelial dysfunction， lipoproteins accumulation and inflammatory cell infiltration， with a complex pathological mechanism during its evolution. Inflammatory response runs through the whole process of occurrence and development of AS， and is a common link of many pathological factors. Intervention of inflammatory response is an effective way and important strategy to control AS. By virtue of its unique advantages， traditional Chinese medicine （TCM） plays an important role in the clinical treatment. According to the TCM theory， the formation and development of AS are closely related to five factors， namely wind， fire， phlegm， blood stasis and deficiency. Among them， blood stasis and turbid phlegm are the most closely related. Blood stasis and turbid phlegm are both pathological products and pathogenic factors. Their formation is mainly due to the imbalance of Yin and Yang in the whole body， dysfunction of viscera and organs and obstruction of transfusion of Qi， blood and body fluid. The pathogenic factors are not only independent， but also related. In recent years， there are more and more studies on the intervention effect and mechanism of TCM on AS， but the intervention effect of TCM on the inflammatory response signaling pathways has not been reported. In this paper， it mainly introduces the understanding of the effect of TCM against AS and seven signaling pathways relating to the intervention effect of TCM on the inflammatory response， which are Janus kinase/signal transduction and activator of transcription （JAK/STAT） signaling pathway， nuclear factor-κB （NF-κB） signaling pathway， mitogen-activated protein kinase （MAPK） dependent signaling pathway， reactive oxygen species （ROS） dependent signaling pathway， CD40-CD40L signaling pathway and Toll-like receptor （TLR） dependent signaling pathway.
Keywords：traditional Chinese medicine;atherosclerosis;inflammatory response;signaling pathway;research progress
Abstract：Shengjiangsan was recorded in a medical book Treatise on Cold-induced Disorders and Plague written in the Qing Dynasty. This prescription， consisting of four drugs： Bombyx， cicada slough， Turmeric and Rhubarb， is simple but effective in regulating Qi and eliminating pathogenic factors. In recent years， the researches on the mechanism of this prescription have been continuously expanded and deepened. According to the collected data， Shengjiangsan is widely used in the treatment of various clinical diseases. The concept of systemic inflammatory response syndrome （SIRS） was first proposed in the 1990s， and it refers to the pathophysiological state caused by self-amplified cascade inflammatory response and inflammatory factor storm arising from infectious or non-infectious factors. In recent years， the treatment of SIRS has gradually become the current focus and hotspot. Patients with severe acute respiratory syndrome （SARS） and novel coronavirus pneumonia （COVID-19） often die from SIRS progression to multiple organ dysfunction syndrome （MODS） or even multiple organ failure （MOF）. Interleukin -1 receptor antagonist， anti-bradykinin， platelet activating factor receptor antagonist and other commonly used drugs in clinical treatment of SIRS have not achieved satisfactory results， so it is urgent to find drugs to improve SIRS status. Shengjiangsan is recommended as an initial prescription in traditional Chinese medicine diagnosis and treatment plan for SARS and COVID-19， and its antiviral effect may be related to preventing SIRS from transforming into MODS. Pharmacological studies have shown that Shengjiangsan can regulate the immune imbalance of SIRS to achieve a balance by intervening various inflammatory factors and their signaling pathways. At present， there are few reports on the treatment of SIRS with Shengjiangsan. Therefore， the theoretical analysis of Shengjiangsan and its clinical research and pharmacological mechanism in the treatment of SIRS are discussed here to broaden the thinking for clinical practice and experimental research.
Abstract：Alzheimer's disease（AD） is an incipient aging neurodegenerative disease， which increases rapidly along with the development trend of social aging and seriously threatens the health of the people. In the absence of effective preventive measures， it will have an enormous impact on the socio-economic and healthcare system. The study found that abnormal cell signal transduction is a key link in many diseases. Cell signal transduction theory has been widely used to clarify the essence of traditional Chinese medicine visceral image and the mechanism of traditional Chinese medicine. 'Correlation of Liver and Kidney' is one of the core plates of the theory of 'Correlation of Five Organs'， which is suitable for explaining the pathogenesis of complex diseases and the correlation of multiple syndromes， and guiding the prescription of clinical syndrome. Hei Xiaoyaosan， as the first choice compound for the prevention and treatment of AD based on the theory of "Correlation of Liver and Kidney' in our team， can play the effects of prevention and treatment by soothing liver and nourishing blood， strengthening spleen and tonifying kidney， and promoting brain collaterals and dredging viscerab spirit. Based on the theory of 'Correlation of Liver and Kidney'， this paper expounds the pathogenesis of AD from the perspective of traditional Chinese medicine， and puts forward the methods and ideas of the preventing and treating of AD from Ca2+-calcium/calmodulin dependent protein （CaM）/calcium/calmodulin dependent protein kinaseⅡ（CaMKⅡ）-cyclic adenosine phosphate reactive element binding protein （CREB） cell signal transduction pathway by consulting literatures and previous studies.
Keywords：correlation of liver and kidney;Alzheimer's disease;Ca2+- calcium/calmodulin dependent protein （CaM）/calcium/calmodulin dependent protein kinaseⅡ （CaMKⅡ）-cyclic adenosine phosphate reactive element binding protein （CREB） signal pathway;Hei Xiaoyaosan
Abstract：Melasma， as a kind of melanosis， often occurs over the face of young and middle-aged women， which seriously affects the physical and mental health of patients. At present， it is believed that the occurrence of melasma is related to various factors such as ultraviolet radiation and changes in sex hormone levels in the body. However， the exact pathogenesis of melasma is still unclear and its clinical efficacy is not ideal. Therefore， it is of great clinical significance and social value to carry out basic researches on melasma diseases. Among them， the animal model of melasma acts as an important tool for studying melasma， and it is particularly important to clarify the pathophysiological mechanism of the occurrence and development of melasma. The common modeling methods include ultraviolet modeling， progesterone modeling and ultraviolet combined with progesterone modeling. However， there are still some problems in the practical application of animal models of melasma due to many influencing factors in the preparation of such animal models， and there is still a lack of a more complete and recognized model preparation scheme to this day， which reduces the success rate of model preparation and limits its application and popularization to some extent. In view of the key problems in the establishment and application of animal models of melasma， we comprehensively summarized the research status of the models from such aspects as experimental animals， modeling methods and model evaluation， and discussed the effects of different modeling methods and animal species， animal age and other factors on animal model of melasma by referring to and sorting out the recent literatures at home and abroad in recent years. This is to provide references for the preparation of more scientific， reasonable， economic and convenient animal models of melasma， lay a foundation for in-depth researches on the pathogenesis， prevention and treatment of melasma， and also provide reference for other animal model research.
Abstract：Polygalae Radix， a traditional Chinese medicine， has the functions of improving intelligence， calming nerves， relieving cough and eliminating phlegm. Its processing methods are various， but the purpose of processing is to reduce toxicity and increase efficiency. In this paper， the methods of ancient processing， such as cleansing， cutting， processing with excipient and processing without excipient， were summarized， the processing methods of Polygalae Radix in the different versions of Chinese Pharmacopoeia and the local processing specifications were summarized， in order to compare the differences and research progress of different processing methods. On this basis， taking the modern research of processed products of Polygalae Radix as the breakthrough point， this paper reviewed the modern research on processed products of Polygalae Radix from the aspects of processing technology， chemical composition changes and pharmacodynamics changes before and after processing， and the mechanism of reducing toxicity and increasing efficiency. Based on the research status of processing of Polygalae Radix， some existing problems were analyzed in this paper， including not many ancient processing methods used in modern times， lack of standardized research on processing technology， few studies on the ingredients introduced by excipients， etc. The author thinks that it is necessary to strengthen the research on the ancient processing of Polygalae Radix combined with processing methods with local characteristics. While discussing the processing technology， combining with the composition and efficacy， we should carry out in-depth research on the processing mechanism of different processing products of Polygalae Radix， so as to provide scientific basis for the rationality of processing of Polygalae Radix and ensure the clinical safety of medication.
Keywords：Polygalae Radix;processing method;chemical composition;pharmacodynamic effect;reducing toxicity and increasing efficacy;processing mechanism;compatibility
Abstract：Clinically， the incidence and mortality of malignant tumors are relatively high， especially in underdeveloped regions or countries. In recent years， with the continuous improvement of people's health awareness， living standards and medical standards， the incidence and mortality of malignant tumors have been declining. At present， malignant tumors are mostly treated by western medicine therapies in clinic， such as surgical resection or radiation therapy， chemical drug therapy， targeted drug therapy and immunotherapy. However， patients with postoperative tumors are prone to relapse and metastasis， with severe adverse reactions and a poor prognosis. And drug resistance and other issues have a serious impact on clinical efficacy and the quality of life of patients. Traditional Chinese medicine believes that malignant tumors belong to the "accumulation" and "abdominal mass"， with both internal and external etiologies. The internal etiology is mainly the insufficient anti-pathogenic energy. The external etiology is mainly six exogenous pathogenic factors to the body seven emotional stimulations. Pathogenic factors， such as deficiencies of Qi and blood， imbalance of Yin and Yang and visceral dysfunction， which lead to the occurrence of malignant tumors. The pathogenesis is mostly based on the asthenia in origin and access in superficiality. The asthenia in origin is mainly due to the insufficient anti-pathogenic energy， with Qi stagnation， blood stasis， phlegm coagulation， and toxic knot as the symptoms. For malignant tumors， like modules， the method for softening hardness to dissipate stagnation is the first choice. Chinese herbal medicine for softening hardness to dissipate stagnation is widely used for malignant tumors in clinic， with a remarkable clinical efficacy. Therefore， in recent years， anti-tumor mechanism and clinical studies of Chinese herbal medicine for softening hardness to dissipate stagnation have become a hotspot at home and abroad. This paper combines the domestic and foreign literatures of the effect of Chinese herbal medicine for softening hardness to dissipate stagnation in treating malignant tumors in both pharmacological trials and clinical research over the past cade. The progress of the studies is reviewed， in the expectation of providing a reference for the clinical anti-tumor application of Chinese herbal medicine for softening hardness to dissipate stagnation.
Keywords：malignant tumor;softening hardness to dissipate stagnation;Chinese herbal medicine;Chinese medicine
Abstract：To review the research progress of anticancer realgar preparations from the perspective of patent，in order to provide a reference for the research of new antitumor realgar drugs. IncoPat database was retrieved with keywords plus IPC classification number. Original data and 72 patents of anti-cancer realgar preparations were obtained and included after artificial denoising. The quantitative analysis was made on the information about application trends，application types and patentee. The technical points of representative patents were summarized. ① The patent types of anticancer realgar preparations are mainly product patents. Its technological innovation has undergone the development of realgar compound agents，arsenic sulfide single agents，nanometer products of realgar agents，bioleaching of realgar preparations，compound realgar extract preparations，compound nano realgar preparations and arsenic sulfide polymorphic crystalline structure. ② Nano realgar agents， realgar bioleaching preparations， new crystalline type realgar preparations and realgar compound preparations. ③ The following studies should be strengthened in the future，involving the comparison of biological effects and toxicity of nano realgar preparations of different preparation processes，the selection of optimal particle size，the druggability of realgar bioleaching preparations，the discovery of new As4S4 crystalline forms and the secondary development of anticancer realgar preparations.