Abstract：ObjectivePowders and decocted powders account for about 1/3 in the Catalogue of Ancient Famous Classical Formulas （the First Batch）， and have a very important position. Determination of preparation technology and particle size in the pulverization process is the key step in the research and development of powders and decocted powders following the original methods. However， there are many terms describing the preparation technology and particle size of powders and decocted powders in ancient Chinese medical books， and the parameters are not clear. Due to the lack of unified basis of particle size， the existing research results have not formed a uniform consensus. Based on ancient textual researches and experimental results， this article discusses the particle size of decocted powders and powders.MethodThrough textual researches of the preparation technology and particle size of powders and decocted powders and powder classification in the 2020 edition of Chinese Pharmacopoeia， the specifications of pulverized particle size were suggested. In addition， Xiebaisan and Danggui Buxuetang were taken as examples to investigate the influence of different particle sizes （4， 10， 24 mesh） on the preparation process of decocted powders and the obtained decoction.ResultThe particle size of 4 mesh was equivalent to that of ancient as big as hemp bean. The contents of index components in Xiebaisan and Danggui Buxuetang with particle size of 4 mesh were higher than that of 10 mesh and 24 mesh， but the particle size of 50 mesh was too fine to be filtered.ConclusionThe suggested particle sizes of powders and decocted powders are recommended as Cumo is the power through 10-mesh sieve， Mo is the power through 24-mesh sieve， Ximo is the power through 80-mesh sieve， as big as hemp bean is the power through 4-mesh sieve and not through 10-mesh sieve.
Abstract：Based on the ancient Chinese medical literature and modern clinical application literature， Mahuangtang was systematically analyzed， including its origin， composition， processing requirements and others， in order to provide support for the development of this famous classical formula. It was found that there were some doubts about Mahuangtang， such as the original variety， the processing of decoction pieces， and the dose conversion ratio. On the basis of fully considering the actual needs of the development of famous classical formula preparations and the usage habits of modern clinical practice， the development suggestions of the author were as follows：Ephedrae Herba should be selected Ephedra sinica without removing knots， Cinnamomi Ramulus should be selected from Cinnamomum cassia， Glycyrrhizae Radix et Rhizoma should select Glycyrrhiza uralensis and be roasted with honey， and Armeniacae Semen Amarum should be selected as Dan-processed products. The total dose of Mahuangtang was 24 g after conversion according to 1 Liang equal to 3 g as following the transition phenomenon and law. The preparation and usage method was as follows：took 1.8 L of water， first added Ephedrae Herba in water for decocting， evaporated 400 mL of water， removed the upper foam， then added the other three drugs， cooked over gentle heat to 500 mL， filtered the residue， took three times a day， about 160 mL warm decoction once. The major function recorded of Mahuangtang in ancient books was exterior sthenia syndrome of exogenous wind cold， it often used in the clinical treatment of cold， acute bronchitis， bronchial asthma and other diseases through the flexible application of the later doctors.
Keywords：famous classical formulas;Mahuangtang;historical evolution;prescription research;processing;traditional Chinese medicine （TCM）;clinical practice of TCM
Abstract：ObjectiveTo observe the clinical efficacy of Xiao Qinglongtang on chronic heart failure with cold phlegm in lung， and explore its mechanism of action.MethodA total of 87 patients with definite chronic heart failure were divided into observation group and control group by random number table method．The two groups received routine western medicine at the same time. Forty-two cases in observation group were treated with Xiao Qinglongtang based on western medicine， and 45 cases in control group received the same dose of placebo. Both groups were treated for 4 weeks， and then their cardiac function， traditional Chinese medicine （TCM） syndrome score and efficacy were compared before and after treatment. Left ventricular ejection fraction （LVEF） and N-terminal pro-brain natriuretic peptide （NT-proBNP） were measured in both groups before and after treatment. The changes of standard deviation of NN intervals （SDNN）， square root of the mean squared differences of successive NN intervals （RMSSD）， high frequency （HF） and low frequency （LF） that reflect autonomic nerve function indexes in heart rate variability （HRV） after treatment were compared between two groups. The changes of inflammatory indicators such as interleukin-6 （IL-6） and highly sensitive C reaction protein （hs-CRP） were detected.ResultAfter treatment， the total effective rate for cardiac function in observation group was higher than that in control group （P<0.05）. The TCM symptom scores were improved after treatment in both groups （P<0.05）， and the total effective rate in observation group was higher than that in control group （P<0.01）. After treatment， LVEF levels significantly increased （P<0.01） and NT-proBNP levels significantly decreased （P<0.01） in both groups， and the effect in observation group was more obvious （P<0.01）. After treatment， SDNN， RMSD， HF and LF indicators in HRV were all higher than those before treatment in both groups （P<0.01）， and the improvement in observation group was more significant than that in control group （P<0.01）. The levels of IL-6 and hs-CRP decreased after treatment in both groups （P<0.01）， and the level of observation group was significantly lower than that of control group （P<0.01）.ConclusionXiao Qinglongtang has certain clinical efficacy in treating chronic heart failure with cold phlegm in lung as it can improve the clinical symptoms of patients， regulate autonomic nervous balance， and inhibit inflammatory factors， providing new clinical ideas to treat chronic heart failure in TCM.
Keywords：Xiao Qinglongtang;chronic heart failure;cold phlegm in lung;heart rate variability;inflammatory factors
Abstract：ObjectiveTo study the effect of Baihutang on blood glucose， blood lipid metabolism and vascular remodeling in type 2 diabetic rats and its regulation on insulin receptor substrate-1（IRS-1）/ phosphatidylinositol-3 kinase（PI3K）/ protein kinase B（Akt） signal pathway.MethodThe 90 rats were randomly divided into normal group， model group， Baihutang low， middle and high dose groups and metformin group， with 15 rats in each group. Except for normal group， the other rats were injected intraperitoneally with streptozotocin to establish the model of type 2 diabetes. The rats in the low， middle and high dose groups were given Baihutang formula granules of 5， 10， 20 g·kg-1 respectively according to their body weight. The positive control group was given metformin （100 mg·kg-1） by intragastric administration， while those in the control group and model group were given the same amount of normal saline once a day for 12 weeks. The levels of fasting blood glucose， glycosylated hemoglobin， serum tumor necrosis factor-α（TNF-α）， interleukin-6（IL-6）， interleukin-1 β（IL-1β）， total cholesterol（TC）， triglyceride（TG） and low-density lipoprotein cholesterol（LDL-C） were measured after administration. The levels of sterol regulatory element binding protein 1C （SREBP1C）， acetyl CoA carboxylase （ACC）， fatty acid synthase gene （FASN） and carnitine palmitoyl transferase 1A （CPT1A）， acylcoa oxidase 1（ACOX1）， recombinant human acylcoa dehydrogenase （ACADM） mRNA in liver of rats were detected by Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）， Western blot was used to detect the protein levels of IRS-1， PI3K and Akt in liver of rats. Hematoxylin-eosin（HE） staining was used for histopathological examination of rat thoracic aortic vessels. The migration ability of vascular smooth muscle cells in rat thoracic aorta was detected by scratch test.ResultCompared with the normal group， the fasting blood glucose， glycosylated hemoglobin， serum TNF-α， IL-6，IL-1β， TC，TG and LDL-C levels， liver lipid synthesis gene mRNA level and vascular smooth muscle cell migration ability of thoracic aorta in model group were significantly higher than those in normal group （P<0.05）， while fatty acid oxidation gene mRNA level and IRS-1，PI3K，Akt protein level in liver were significantly decreased in model group （P<0.05）. The vascular wall thickness of thoracic aorta increased significantly in rats （P<0.05）. Compared with model group， the levels of fasting blood glucose， glycosylated hemoglobin， serum TNF-α，IL-6， IL-1β， TC， TG and LDL-C， the level of lipid synthesis gene mRNA in liver and the migration ability of vascular smooth muscle cells in thoracic aorta of rats in all Baihutang groups were significantly lower than those in model group （P<0.05）. The mRNA level of fatty acid oxidation gene and the protein levels of IRS-1， PI3K and Akt in liver were significantly increased（P<0.05）， and the histopathology of thoracic aorta was significantly improved and the vascular wall thickness decreased significantly（P<0.05）.ConclusionBaihutang can reduce the levels of blood glucose， blood lipid and serum inflammatory factors in type 2 diabetic rats， regulate the expression of genes related to lipid metabolism in liver， and improve the histopathology and vascular remodeling of thoracic aorta. The mechanism may be related to the regulation of IRS-1/PI3K/Akt signal pathway.
Abstract：ObjectiveTo observe the effect of Shaofu Zhuyutang on nuclear factor erythroid-2-related factor 2 （Nrf2） /antioxidant response element （ARE） signaling pathway in blood vessels by establishing the model of rats with cold coagulation and blood stasis syndrome， and to explore the protective effect and mechanism of Shaofu Zhuyutang on vascular endothelial injury.MethodThe 50 SPF rats were randomly divided into high dose group （4.8 g·kg-1）， middle dose group （2.4 g·kg-1）， low dose group （1.2 g·kg-1）， model group and normal group （ten of each group）. The rat model of cold coagulation and blood stasis syndrome was established by subcutaneous injection of epinephrine hydrochloride combined with ice bath. At the same time of modeling， the drug was administered by gavage. After 28 days of continuous administration， the hemorheology indexes were detected by automatic hemorheology instrument. Levels of nitric oxide （NO）， endothelin （ET）-1， superoxide dismutase （SOD）， glutathione （GSH-Px）， intercellular adhesion molecule-1（ICAM-1）， vascular cell adhesion molecule-1（VCAM-1）， von Willebrand factor （vWF） in serum were determined by ELISA. Hematoxylin and eosin （HE） staining was used to observe the endothelial injury of vascular tissue of thoracic aorta. The protein expression of Nrf2 and HO-1 in vascular tissue of thoracic aorta was detected by Western blot. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）was used to observe the expression of Nrf2 and heme oxygenase-1 （HO-1） mRNA in vascular tissue of thoracic aorta.ResultCompared with the blank group， model group rats whole blood viscosity and plasma viscosity were significantly increased （P<0.05，P<0.01）， vWF， ICAM 1， VCAM 1 content increased significantly （P<0.01）， NO， SOD， gsh-px levels decreased significantly （P<0.01）， significantly increased the content of ET-1（P<0.01）， thoracic aorta vascular tissue Nrf2， HO-1 mRNA expression was significantly increased （P<0.01）， Nrf2 protein expression in the cell nucleus increased significantly （P<0.05）， The protein expression level of Nrf2 in cytoplasm was significantly decreased （P<0.05）， while the protein expression level of HO-1 was significantly increased （P<0.01）. Compared with model group， the whole blood viscosity （high and middle cut）， plasma viscosity， were significantly reduced in high and meduim-dose Shaofu Zhuyutang groups（P<0.05，P<0.01）. The levels of vWF， ICAM-1， VCAM-1 and ET-1 in serum were significantly reduced （P<0.05，P<0.01）， NO， SOD and GSH-Px increased significantly （P<0.05，P<0.01）. The pathological changes such as hyperplasia， swelling and shedding of endothelial cells of thoracic aorta， rupture of internal elastic membrane and disorder of smooth muscle arrangement were improved. The expression levels of Nrf2， HO-1 protein and gene were significantly increased in vascular tissue of thoracic aorta （P<0.01）.ConclusionShaofu Zhuyutang has a protective effect on vascular endothelial injury in rats with cold coagulation and blood stasis syndrome. The mechanism of action is related to the activation of Nrf2/ARE signaling pathway， which leading to the increased expression of antioxidant enzymes and decreased the expression of adhesion factors.
Keywords：Shaofu Zhuyutang;endothelial cells injury;nuclear factor erythroid-2-related factor 2 （Nrf2） /antioxidant response element （ARE）;cold coagulation and blood stasis syndrome
Abstract：ObjectiveTo study the effect of Biejiajian Wan on the epithelial-mesenchymal transition （EMT） of rat hepatic oval cells induced by transforming growth factor- β1（TGF-β1）， in order to explore its mechanism in reversing EMT.MethodWB-F344 cells were divided into five groups： blank group， TGF-β1 model group （10 μg·L-1TGF-β1）， low-dose group （10 μg·L-1TGF-β1+0.55 g·kg-1Biejiajian Wan）， medium-dose group （10 μg·L-1TGF-β1+1.1 g·kg-1Biejiajian Wan）， high-dose group （10 μg·L-1TGF-β1+2.2 g·kg-1Biejiajian Wan）. Except blank group， TGF-β1 was used to induce WB-F344 cells in all of the remaining groups to construct an EMT model. After the cells were treated with low， medium and high doses of Biejiajian Wan serum， the changes of migration ability of WB-F344 cells were detected by cell scratching test. The expressions of E-cadherin， N-cadherin and Vimentin were detected by Western blot. Real-time PCR was used to detect the changes in the expression of β-catenin mRNA. The expression of β-catenin was detected by cell immunofluorescence assay.ResultCompared with normal WB-F344 cells， the intercellular space of WB-F344 cells became loose from tight， and the morphology changed from cobblestone to fibroblast after TGF-β1 induced WB-F344 cells for 4 days， and the expression of E-cadherin protein decreased， while the expression of N-cadherin protein increased （P<0.01）， indicating that the EMT model of WB-F344 cells was successfully built. Compared with the blank group， the migration ability of WB-F344 cells in TGF-β1 model group was enhanced （P<0.01）， compared with TGF-β1 model group， Biejiajian Wan could significantly inhibit the migration ability of WB-F344 cells； specifically， the low-dose group had no statistical significance， and the medium and high-dose groups had statistical significance （P<0.05）. Western blot results showed that compared with the blank group， the expression of E-cadherin decreased， whereas those of N-cadherin and Vimentin increased in the TGF-β1 model group （P<0.01）， compared with TGF-β1 model group， E-cadherin protein expression was increased in the low， medium and high-dose groups， while the expressions of N-cadherin and Vimentin was decreased； specifically， the low-dose groups had no statistical significance， and the medium and high dose groups had statistical significance （P<0.05，P<0.01）. Real-time PCR results showed that compared with the blank group， the mRNA expression of β-catenin in the TGF-β1 model group was increased （P<0.05）， whereas compared with TGF-β1 model group， the mRNA expression of β-catenin in the low， medium and high-dose groups of Biejiajian Wan was reduced （P<0.01）. The results of cellular immunofluorescence showed that compared with the blank group， the fluorescence expression of β-catenin in the cell nucleus was enhanced in the TGF-β1 model group； and compared with the TGF-β1 model group， the expression of β -catenin in the cell nucleus of the low， medium and high-dose groups of Biejiajian Wan decreased， and the inhibitory effect of Biejiajian Wan on β-catenin in the cell nucleus was positively correlated with its concentration.ConclusionBiejiajian Wan may reverse the EMT process that TGF-β1 induced WB-F344 cells， and inhibit the migration of WB-F344 cells by inhibiting Wnt/β-catenin signaling pathway.
Abstract：Alzheimer's disease （AD） is a neurodegenerative disease that leads to progressive memory and cognitive impairment and behavioral disorders， which has seriously threatened the health of the majority of middle-aged and elderly people. Traditional Chinese medicine （TCM） believes that the basic pathogenesis of AD is deficiency of kidney-essence， blood stasis and meridian stagnation. In recent years， many studies have shown that TCM has obvious value and advantages in the prevention and treatment of AD by multi-target mechanism. Therefore， it is of great significance to screen out effective anti-AD drugs from TCM compound prescriptions. Huangjingwan， also known as Jiuzhuan Huangjingwan， has the effects in tonifying kidney-essence， activating blood and removing stasis， with a potential effect in preventing AD. In this article， the feasibility of Huangjingwan in the prevention and treatment of AD was analyzed and discussed from the perspective of TCM theory， the study results of Huangjingwan in the prevention and treatment of AD were summarized， and the mechanism of its action was analyzed from the perspective of pharmacological mechanism. Based on TCM theory， Huangjingwan has the effect of anti-AD. According to relevant findings， Huangjingwan has many targets， such as anti-oxidation， anti-inflammatory， decrease of the level of oxidative stress in brain， activation of Wnt/β-catenin signal transduction in brain， regulation of glycogen synthase kinase-3β （GSK-3β）， protein phosphatase 2A （PP2A） activity balance， reduction of amyloid β （Aβ） content and tau protein hyperphosphorylation in brain， so as to exert effects in improving neurological symptoms and increasing learning and memory ability， with an anti-AD neuroprotective function. This will provide new ideas for in-depth studies and clinical applications of Huangjingwan against AD.
Abstract：ObjectiveTo treat mice with Alzheimer's disease （AD） with β-catenin RNA interference （RNAi） Huangjingwan （HW）， so as to explore the neuroprotective signal mechanism of its prevention and treatment of AD.MethodA total of 81 male Kunming mice were randomly divided into normal control group， sham model control group， AD model group， Donepezil group， HW+scrambled group， HW+RNAi group， HW group， with 8 mice in each of donepezil group and HW group， and 13 mice in each of other groups. The AD models were established through injection with D-galactose and scopolamine in the last 5 groups for 5 consecutive weeks. On the 1st day of the 4th week after modeling， 0.75 μL PEI-LMW/β-catenin siRNAs nano-complex was injected into the right lateral ventricle of each mouse in for one time to treat with β-catenin RNAi in mice brains of the HW+RNAi group. The 0.75 μL complex was injected into the right lateral ventricle of each mouse for one time as for β-catenin interference control of the HW+scrambled group. The 0.75 μL normal saline was injected into the right lateral ventricle of each mouse in one time of the sham control group. Two weeks after intracerebroventricular injection， β-catenin RNAi was confirmed to be successful， and Donepezil （6.5×10-4 g·kg-1） was intragastrically administered to each mouse of donepezil group. HW （2.5 g·kg-1） was intragastrically administered to each mouse of HW group， HW+RNAi group and HW+scrambled group. Normal saline （0.5 mL·d-1） was intragastrically administered to each mouse of the sham control group. All gastric perfusion lasted for 4 weeks. At the end of gavage， the difference in learning and memory ability of mice was evaluated by platform jumping test. Nissl staining was used to count the number of neurons in s1Tr area of cerebral cortex and CA1 and CA3 areas of hippocampus of each mouse in each group. The mRNA expressions of Wnt1， DVL2， GSK-3β， β-catenin and CyclinD1 in mice brain of each group were detected by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Western blot was used to detect the expressions of Wnt1， DVL2， GSK-3β， β-catenin and CyclinD1 in mice brain of each group.ResultThe expression of β-catenin could be significantly inhibited through the injection with PEI-LMW/β-catenin siRNAs nano-complex into the lateral ventricle of AD mice， and nearly no β-catenin expression could be detected， which successfully achieved gene silencing. Compared with the normal control group， mice in AD model group showed that the learning and memory performance decreased significantly， the number of jumping errors increased （P<0.01）， the number of neurons in S1Tr area of cerebral cortex and CA1， CA3 areas of hippocampus decreased significantly （P<0.01）， the mRNA and protein expressions of Wnt1， DVL2， β-catenin， CyclinD1 in brain decreased significantly （P<0.01）， while the mRNA and protein expressions of GSK-3β increased significantly （P<0.01）. Compared with the AD model group， mice in HW group showed that the learning and memory performance increased significantly， the number of jumping errors decreased， the number of neurons in S1Tr area of cerebral cortex and CA1， CA3 areas of hippocampus increased significantly， the mRNA and protein expressions of Wnt1， DVL2， β-catenin， CyclinD1 in brain increased significantly， while the mRNA and protein expression of GSK-3β decreased significantly （P<0.01）. Compared with the HW group， mice in HW+RNAi group showed that the learning and memory performance decreased significantly， the number of jumping errors increased significantly （P<0.01）， the number of neurons in S1Tr area of cerebral cortex and CA1， CA3 areas of hippocampus decreased significantly （P<0.01）， there was no significant change in mRNA and protein expressions of Wnt1， DVL2， GSK-3β in the brain， and the mRNA and protein expressions of β-catenin， CyclinD1 decreased significantly （P<0.01）.ConclusionHW can treat and prevent AD by activating Wnt/β-catenin signal pathway.
Keywords：RNA interference （RNAi）;Huangjingwan;Alzheimer's disease;Wnt/β-catenin signal pathway
Abstract：ObjectiveTo explore the effect of Huangjingwan （HW） on the expressions of Wnt/β-catenin signal pathway-associated proteins in the hippocampus of mice with Alzheimer's disease （AD） induced by D-galactose and okadaic acid with learning and memory disorders， as well as its mechanism.MethodAfter subcutaneous injection with 1.0% D-galactose （0.14 g·kg-1·d-1） into the back and neck of mice for 4 weeks， the right ventricle of mice was injected with 2 μL（75 ng） of okadaic acid for one time to make AD model， and the successfully modeled AD mice were selected by Morris water maze. Then， the selected AD mice were randomly divided into AD model group， memantine group （1.3×10-3 g·kg-1·d-1） and HW group （2.5 g·kg-1·d-1）. In addition， the sham model control group and the normal control group were set up. At the same time， 2 μL normal saline was injected into the right ventricle of mouse in the sham model control group as the modeling control. Two weeks after molding， the mice in the two experimental drug groups were given the corresponding dose of the experimental drug by gavage for 4 weeks. In addition， after 2 weeks of AD modeling， mice in sham model control group and AD model group were intragastrically administrated with the same amount of normal saline daily for 4 weeks. There was no special treatment in the normal control group. At the end of gavage， the shuttle experiment was performed to detect the differences in learning and memory levels of mice in each group. The changes of β-catenin and GSK-3β positive neurons in CA1 area of hippocampus in each group were tested by immunohistochemistry. Quantitative real-time polymerase chain reaction （Real-time PCR） was used to measure the mRNA expressions of GSK-3β， β-catenin and CyclinD1 in hippocampus of mice in each group. The Western blot was used to detect the expressions of total GSK-3β （t-GSK-3β）， phosphorylation of GSK-3β at Ser9 （p-Ser9-GSK-3β）， phosphorylation of GSK-3β at Tyr216 （p-Tyr216-GSK-3β）， total β-catenin （t-β-catenin）， phosphorylation of β-catenin （p-β-catenin） and CyclinD1 proteins in hippocampus of mice in each group.ResultCompared with the normal control group， mice in AD model group showed an obvious dementia state， which was characterized by significant declines in learning and memory ability， the number of β-catenin immunoreactive neurons in hippocampal CA1 area， the mRNA and protein expressions of t-β-catenin and CyclinD1， the protein expressions of p-Ser9-GSK-3β， and the ratio of p-Ser9-GSK-3β/t-GSK-3β and p-Tyr216-GSK-3β/t-GSK-3β in hippocampal region （P<0.01）， and significant increases in the number of GSK-3β immunoreactive neurons in hippocampal CA1 area， the mRNA and protein expressions of t-GSK-3β， the protein expressions of p-Tyr216-GSK-3β and p-β-catenin， the ratio of p-β-catenin/t-β-catenin in hippocampal region （P<0.01 respectively）. Compared with the AD model group， the dementia symptoms of mice in HW group were significantly alleviated， and the number of β-catenin immunoreactive neurons in hippocampal CA1 area， the mRNA and protein expressions of t-β-catenin and CyclinD1， the protein level of p-Ser9-GSK-3β， the ratio of p-Ser9-GSK-3β/t-GSK-3β in hippocampal region were all significantly increased （P<0.01 respectively）， whereas the number of GSK-3β immunoreactive neurons in hippocampal CA1 area， the mRNA and protein expressions of t-GSK-3β， the proteins expressions of p-Tyr216-GSK-3β and p-β-catenin， the ratio of p-β-catenin/t-β-catenin in hippocampal region were all significantly decreased （P<0.01 respectively）， but the ratio of p-Tyr216-GSK-3β/t-GSK-3β has no significant statistical difference.ConclusionHW shows the role of AD treatment， which can down-regulate the expression of GSK-3β in the hippocampus of AD mice and reduce its protein activity， and up-regulate the expression of β-catenin as well as increase its protein activity， so as to enhance the expression of downstream CyclinD1 and promote the transcription of the target genes. Its mechanism may be related to the activation of Wnt/β-catenin signal pathway.
Keywords：Huangjingwan;Wnt/β-catenin signal pathway;Alzheimer's disease;hippocampal neurons
Abstract：ObjectiveTo investigate the effect of Huangjingwan （HW） on the activities of glycogen synthase kinase-3β （GSK-3β）， protein phosphatase 2A （PP2A） and the mechanism in inhibiting tau protein hyperphosphorylation in the hippocampal neurons of mice with Alzheimer's disease.MethodAfter subcutaneous injection with 1.0% D-galactose （0.14 g·kg-1·d-1） into the back and neck of mice for 4 weeks， the right ventricle of mice was injected with 2 μL （75 ng） of okadaic acid for one time to make AD model， and the successfully modeled AD mice were selected by Morris water maze. Then， the selected AD mice were randomly divided into AD model group， memantine group （1.3×10-3 g·kg-1·d-1） and HW group （2.5 g·kg-1·d-1）. In addition， the sham model control group and the normal control group were set up. At the same time， 2 μL normal saline was injected into the right ventricle of mouse in the sham model control group for modeling control. Two weeks after modeling， the mice in the two experimental drug groups were given the corresponding dose of the experimental drug by gavage for 4 weeks. In addition， after 2 weeks of AD modeling， mice in control group and AD model group were intragastrically administrated with the same amount of normal saline daily for 4 weeks. The mice in normal control group were only given daily feed. At the end of gavage， all the mice were tested by the open field experiment and jumping platform experiment to evaluate the differences in exploratory activity ability， anxiety level and learning and memory ability. The number of neurons in CA1 and CA3 areas of hippocampus in all the mice was detected by Nissl staining. Quantitative real-time polymerase chain reaction （Real-time PCR） was used to detect mRNA expressions of GSK-3β and PP2A in hippocampus of mice in each group. Protein expressions of GSK-3β， PP2A， phosphorylated tau （p-tau） and total tau protein （t-tau） in hippocampus of mice in each group were detected by Western blot.ResultCompared with the normal control group， mice in AD model group showed an obvious dementia state， which was characterized by a lower spontaneous activity， lower exploration behavior ability， higher anxiety level， less movement and easier to stay and hide， longer learning response time， significantly increased number of learning and memory errors， and decreased numbers of hippocampal neuron in CA1 and CA3 areas， and reduced mRNA and protein expressions of PP2A， mRNA and protein expressions of GSK-3β， p-tau protein and the ratio of p-tau/t-tau were all increased significantly （P<0.01）， while expression of t-tau protein was decreased， with no significant difference. Compared with the AD model group， mice in the HW group showed a higher spontaneous activity， higher exploration ability， lower anxiety level， higher learning and memory performance， and the numbers of hippocampal neuron in CA1 and CA3 areas increased， while mRNA and protein expressions of PP2A increased， and the mRNA and protein expressions of GSK-3β， the expression of p-tau protein and the ratio of p-tau/t-tau were all decreased significantly （P<0.01）， but with no significant difference in the protein expression of t-tau.ConclusionHW can inhibit tau hyperphosphorylation in hippocampal neurons of AD mice， restore tau protein function， protect hippocampal neurons， and exert an anti-AD effect， which may be related to the regulatory mechanism in the activity balance between GSK-3β and PP2A in hippocampal neurons.
Abstract：ObjectiveTo explore the effect and mechanism of Guizhitang with different proportions of Cinnamomi Ramulus and Paeoniae Alba Radix on overactive cardiac sympathetic nerves in salt-sensitive hypertensive rats.MethodRandomly divide the forty male 6-week-old salt-sensitive hypertensive rats into five groups： the normal control group， the model group， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶2 group，and the Cinnamomi Ramulus and Paeoniae Alba Radix 2∶1 group， each group has 8 animals， the normal control group was fed with low-salt feed， and the remaining four groups were fed with 8% high-salt feed. After 4 weeks of feeding， gastric feeding was started. Give both the normal control group and model group saline and the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group， the 1∶2 group，and the 2∶1 group， were given Guizhitang aqueous solution at 4.0， 5.5 and 5.5 g·kg-1， respectively. Continuous gavage intervention was held for 4 weeks. IITC multi-channel non-invasive sphygmomanometer was used to detect changes of systolic blood pressure before and after treatment in rats. Left ventricular anterior wall end-diastolic thickness （LVAWd） and interventricular septal diastolic thickness （IVSd） were detected by echocardiography. Hematoxylin-eosin（HE）staining and Masson staining were used to observe the myocardial morphological changes of rats in each group， Western blot was used to detect the expression of nerve growth factor （NGF）， growth-associated protein 43 （GAP43） and tyrosine hydroxylase （TH） protein.ResultAfter 4 weeks of intervention with Guizhitang， compared with the normal control group， the blood pressure， LVAWd and IVSd of the model group were significantly increased， and the expressions of NGF， TH and GAP43 protein were significantly increased （P<0.01）. HE and Masson staining results showed that the model group had myocardial cell edema， a large number of inflammatory cell infiltration， myocardial fiber hyperplasia and disordered arrangement， and a large amount of collagen deposition could be seen in the intercellular substance. Compared with model group， the systolic blood pressure of rats in each Guizhitang group increased slowly， and the expression of NGF， TH and GAP43 protein decreased （P<0.05，P<0.01），the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group has the best effect. The results of echocardiography shows that the 1∶1 Guishao group could reduce LVAWd and IVSd levels （P<0.05，P<0.01）， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶2 group could reduce IVSd level （P<0.05）， there was no statistical difference in LVAWd， there was no statistical difference in LVAWd and IVSd in 2∶1 group. In terms of myocardial morphology， each group of Guizhitang can reduce cell edema and inflammatory cell infiltration， reduce myocardial fiber hyperplasia and collagen deposition， and improve the disorder of myocardial fiber arrangement. Among them， the 1∶1 group has the best effect.ConclusionGuizhitang can inhibit the overactive activation of cardiac sympathetic nervous system，reduce the extent of myocardial fibrosis， inflammatory infiltration and myocardial hypertrophy， and protect salt-sensitive hypertension rats， whose mechanism may be related to regulating the expression of heart NGF.Among them， the Cinnamomi Ramulus and Paeoniae Alba Radix 1∶1 group is better than the 1∶2 and 2∶1 group in reducing myocardial fibrosis， inflammatory infiltration and myocardial hypertrophy.
Abstract：ObjectiveTo study the effect of Shuangshen Xionglian （SSXL） granules on vasculopathy and phosphatidylinositol 3 kinase （PI3K）/serine threonine kinase （Akt）/nitrogen oxide synthase （eNOS） signal in hyperhomocysteinemia chronic kidney disease rats.MethodRats were randomly divided into 5 groups： sham operation group， model group， and high， medium and low-dose （8， 4， 2 g·kg-1） SSXL groups. The model of hyperhomocysteinemia chronic kidney disease in rats was established with high methionine feed combined with 5/6 nephrectomy. After 5/6 nephrectomy， continuous intragastric administration lasted for four weeks. Arterial blood pressure was measured at the 4th and 8th weeks after operation. At the end of the 8th week after the operation， blood was collected to determine serum creatinine， urea nitrogen， homocysteine （Hcy）， methionine and blood lipid. Western blot was used to detect the expressions of PI3K/Akt/eNOS pathway-related proteins， such as p-p85， p-Akt and p-Ser177 in thoracic aorta， and serum NO and eNOS were measured. The changes of endothelium-dependent relaxation and non-endothelium-dependent relaxation were measured by the method of isolated thoracic aorta ring. Pathological htoxylin eosin （HE） staining was used to observe the changes of renal tissue and thoracic aorta.ResultAt the 8th week of the experiment， compared with the sham operation group， arterial systolic blood pressure， serum urea nitrogen， creatinine， Hcy， methionine， total cholesterol and low-density lipoprotein of the model group were significantly increased. Four weeks later after administration， arterial systolic blood pressure， serum urea nitrogen， Hcy， methionine， serum total cholesterol and serum low-density lipoprotein were significantly reduced in each dose group （P<0.05， P<0.01）. The creatinine in the SSXL 8， 4 g·kg-1 group was significantly reduced （P<0.05）. The nitric oxide content of SSXL in each dose groups were increased compared with that in the model group （P<0.05， P<0.01）， and the serum eNOS activity of the SSXL in the SSXL 8 g·kg-1 group was significantly increased compared with that in the model group （P<0.05）. The endothelium dependent and non-endothelium dependent vasodilation of thoracic aortic rings in the model group were significantly damaged. The cumulative concentration of acetylcholine （1×10-5.5~1×10-4 mmo1·L-1） in the SSXL 8 g·kg-1 group was significantly improved （P<0.05， P<0.01）. The diastolic degree of the vascular ring in the SSXL 8 g·kg-1 group was significantly higher than that in the model group （P<0.05）. Western blot results showed that the expressions of p-85， p-Akt and p-Ser177 in blood vessels increased in the sham group compared with those in the model group （P<0.01）. Compared with the model group， the phosphorylation level of this pathway was increased in the SSXL groups， and the expressions of p-Akt and p-Ser177 in the SSXL 8 g·kg-1 group were significantly increased （P<0.05）. The pathological results showed that the pathological changes of thoracic aorta and renal tissue in the dosages of SSXL were significantly reduced compared with those in the model group.ConclusionSSXL granules can improve hyperhomocysteine and dyslipidemia in rats of chronic kidney disease with hyperhomocysteine， reduce serum creatinine， urea nitrogen levels and arterial systolic blood pressure， and improve vascular morphology and diastolic function， which may be related to the regulation of the PI3K/Akt/eNOS signaling pathway.
Abstract：ObjectiveTo observe the effect of Qiyu Sanlong decoction （QYSL） on the expressions of key molecules in signal axis of mammalian rapamycin target protein （mTOR）/yeast Atg6 homologous （Beclin1）/ microtubule-associated protein1 light chain3 （LC3） in A549 cells.MethodWith A549 cells as the research object， the effect of QYSL medicated serum on cell viability of A549 cells were detected by cell counting kit-8 （CCK-8） method. The effect of QYSL decoction on A549 cell apoptosis， autophagosome formation and the expression of autophagy markers were detected by Terminal-deoxynucleoitidyl transferase mediated nick end labeling （TUNEL） method， transmission electron microscope （TEM）， Real-time polymerase chain reaction （Real-time PCR） and Western blot.ResultQYSL medicated serum could inhibit the viability of A549 cells in a concentration-dependent manner. Compared with the blank serum group， the number of apoptotic A549 cells in the QYSL medicated serum group was significantly increased （P<0.01）， and the formation of autophagosome was significantly increased. Compared with the blank serum group， the mRNA and protein expressions of mTOR in A549 cells in the QYSL serum group were significantly decreased （P<0.01）， while mRNA and protein expressions of Beclin-1， autophagy related genes 5 （ATG5）， autophagy related genes 13 （ATG13） were significantly increased （P<0.01）.ConclusionQYSL decoction can induce autophagy in A549 cells， and its specific mechanism may be related to the down-regulation of mTOR expression， the up-regulation of Beclin1， ATG5， ATG13 and LC3 expression， and the promotion of LC3Ⅰ conversion to LC3Ⅱ.
Abstract：ObjectiveTo observe the effect of forsythiaside A on gastrointestinal motility disorder induced by chemotherapy in mice， and explore the mechanism of forsythiaside A regulating gastrointestinal motility.MethodThe 60 KM mice were randomly divided into normal group， model group， metoclopramide group （5 mg·kg-1） and forsythiaside A low， medium and high-dose groups （30， 60， 120 mg·kg-1）， 10 for each group， which include half male and half female. The above dose was given once a day for 4 consecutive days， which the intragastric volume was 10 mL·kg-1. One hour after 1rd day administration， equal volume of saline was intraperitoneally injected to the normal group， 2 mg·kg-1 cisplatin was intraperitoneally injected to the other groups with daily for 4 consecutive days. Observing the effects of forsythiaside A on gastric emptying and small intestinal propulsion on mice models， serum gastrin （GAS） and somatostatin （SS）， motilin （MTL）， vasoactive intestinal peptide （VIP） levels were examined by enzyme-linked immunosorbent assay （ELISA）. Activities of acetylcholinesterase （AChE） and total nitric oxide synthase （tNOS） in gastric antrum and ileum were detected by ELISA. The expression of AChE and inducible nitric oxide synthase （iNOS） in gastric antrum and ileum were detected by Western blot.ResultCompared with normal group， the gastric retention rate and small intestinal propulsion rate of the model group were significantly increased （P<0.01）， serum levels of MTL， GAS， SS and VIP， the AChE activity in the homogenate of ileum in the model group were significantly reduced （P<0.05，P<0.01）， while the tNOS activities in gastric antrum and ileum were significantly increased （P<0.05，P<0.01）. Protein expression of AChE in gastric antrum and ileum were significantly decreased （P<0.05）， and the expression level of iNOS protein was significantly increased in the model group （P<0.05）. Compared with model group， different doses of forsythiaside A can reduce the gastric residual rate and small intestinal propulsion rate of mice to varying degrees. Meanwhile forsythiaside A can increase the serum levels of MTL， GAS， SS， and VIP， and the AChE activity and protein expression levels in gastric antrum and ileum tissues were also increased， while tNOS activity and iNOS protein expression were decreased in gastric antrum and ileum （P<0.05，P<0.01）.ConclusionForsythiaside A can significantly ameliorate the delayed gastric emptying and small intestine hyperfunction induced by cisplatin in mice. Its mechanism to ameliorate gastrointestinal dysfunction caused by chemotherapy is related to the regulation of gastrointestinal AChE and NOS activity in gastric antrum and ileum and the regulation of gastrointestinal hormone levels.
Abstract：ObjectiveTo investigate the effect of modified Si Junzitang on the expression of fibrous protein-5（Fibulin-5）， phosphorylated protein kinase B（p-Akt ）in hippocampus of rats after cerebral ischemia-reperfusion （I/R） injury and the anoikis of nerve cells.MethodThe 60 male SD rats of SPF grade were randomly divided into sham operation group， model group， Edaravone group （3.2 mg·kg-1）and modified Si Junzitang high， medium and low-dose groups（19.08，9.54，4.77 g·kg-1）.The middle cerebral artery occlusion （MCAO） model was established by suture method，the rats were killed 7 days later，neurological deficit score was evaluated before the death，histopathological observation was performed by hematoxylin eosin staining， apoptosis index of nerve cells was detected by TdT-mediated dUTP nick end labeling（TUNEL）staining， the expression of Fibulin-5， p-Akt and protein in ischemic hippocampus were detected by immunohistochemistry and Western blot.ResultThe neurological deficit score showed that，compared with the sham operation group， the neurological deficit score of the model group was significantly increased （P<0.01）， compared with model group， the neurological deficit score of Edaravone group，the high， medium， low dose groups of modified Si Junzitang were decreased （P<0.05，P<0.01）. Immunohistochemical results showed that，compared with the sham operation group， the expression of Fibulin-5， p-Akt protein and the apoptosis index of nerve cells in the model group were significantly increased （P<0.05，P<0.01）， compared with model group， the protein expressions of Fibulin-5 and p-Akt in Edaravone group， high， medium and low-dose groups of modified Si Junzitang were significantly increased （P<0.05，P<0.01）， and the apoptosis index of nerve cells was obvious，there was a significant decrease （P<0.05，P<0.01）. Western blot results showed that，compared with the sham operation group， the relative expression of Fibulin-5 and p-Akt protein in the model group was significantly down-regulated （P<0.01）， compared with model group， the protein expressions of Fibulin-5 and p-Akt in the Edaravone group， the high， medium and low-dose groups of modified Si Junzitang were significantly up-regulated （P<0.05，P<0.01）.ConclusionThe modified Si Junzitang may stabilize the extracellular matrix （ECM） Fibulin-5， increase the adhesion of ECM to cells and promote the expression of p-Akt protein， thus inhibiting neuronal apoptosis and protecting cerebral ischemia injury.
Keywords：modified Si Junzitang;ischemia reperfusion;anoikis;fibrous protein-5（Fibulin-5）;phosphorylated protein kinase B（p-Akt）
Abstract：Objectiveto explore the mechanism of modified Tianwang Buxindan in improving abnormal glucose and lipid metabolism in mice with chronic sleep deprivation from the signal pathway of orexin A/ orexin receptor 1（OX1R）.MethodThe 50 6-week-old male C57BL/6 mice were randomly divided into blank group ， model group ， estazolam group and Tianwang Buxindan low and high dose groups ，for ten mice of each group. Except the blank group， rats were deprived of sleep for 8 weeks by the method of multi-platform water environment. In the last 4 weeks， Tianwang Buxindan （8.5，17 g·kg-1）and estazolam solution（9.1 mg·kg-1）were given to the stomach， and the blank group and the model group were fed with pure water of the same volume. The food intake and body weight of mice were measured twice a week， on the 49th day， blood samples were collected from the tail vein for glucose tolerance test （GTT），on the 52nd day for insulin tolerance test（ITT）， was used to detect the expression of total cholesterol （TCH）， triglyceride（TG）and free fatty acid（FFA）in serum， and enzyme-linked immunosorbent assay（ELISA）was used to detect the expression of orexin A in serum and hypothalamus. Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）and Western blot were used to detect the mRNA and protein expression of OX1R in hypothalamus.ResultAfter administration， the food intake of mice in each group was different， compared with the blank group， the body weight of model group was significantly reduced（P<0.05）， the glucose tolerance was significantly abnormal， and the TCH， TG， FFA values were significantly increased（P<0.01）. The expression of orexin A in serum and hypothalamus increased significantly（P<0.01）， and the mRNA and protein expression levels of OX1R in hypothalamus increased significantly（P<0.01）. Compared with the model group， the body weight of each group of Tianwang Buxindan was significantly increased（P<0.05）， with better glucose tolerance and insulin sensitivity， TCH， TG， FFA values were significantly reduced（P<0.05，P<0.01）， accompanied by serum and the expression of orexin A in the hypothalamus was significantly decreased（P<0.05，P<0.01）， the mRNA and protein expression levels of OX1R were significantly decreased（P<0.05，P<0.01）.ConclusionTianwang Buxindan can protect mice from abnormal glucose and lipid metabolism induced by chronic sleep deprivation， and its mechanism may be related to the down-regulation of orexin A/OX1R signal expression.
Keywords：Tianwang Buxindan;sleep deprivation;glucose and lipid metabolism;orexin A;orexin receptor 1
Abstract：ObjectiveTo observe the efficacy of modified Huanglian Wendantang in treating newly diagnosed type 2 diabetes mellitus （T2DM） with phlegm （dampness）-heat syndrome， in order to study the effect on islet β cell function and adipocytokines.MethodA total of 130 patients were randomly divided into two groups by random number table （65 cases in each group）. The 60 patients in control group completed the treatment （4 patients fell off or lost visit， 2 were eliminated because of breach of plan）， and the 61 patients in observation group completed the treatment （3 patients fell off， 1 were eliminated）. And 20 healthy volunteers were taken as normal control group. Both groups′ patients got lifestyle interventions and metformin hydrochloride tablets （1 tablet/time， 1 time/day during the meal）. In addition， patients in control group got Huazhuo Qingshen Keli in the morning and at night， 5 g/time， 2 times/day， and patients in observation group got modified Huanglian Wendantang， 1 dose/day. And the treatment was lasted for 3 months. Before and after treatment， levels of fasting blood glucose （FBG）， postprandial 2 blood glucose （PBG）， HbA1c and fasting insulin （FINS）， insulin resistance index （HOMA-IR）， insulin sensitivity index （InISI）， islet β cell function index （HOMA-β）， early insulin secretion index （I30/△G30） and late insulin secretion index （AUCI30~I120/G30~G120）， total cholesterol （TC） and triglycerides （TG）， high-density lipoprotein （HDL-C）， low-density lipoprotein （LDL-C）， adiponectin， TNF -α （TNF-α）， resistin and leptin were detected. And syndrome of phlegm （dampness） combined with heat were scored， and the safety was discussed.ResultThe total effective rate in observation group was 91.80% （56/61）， which was higher than 78.33% （47/60） in control group （χ2=4.333， P<0.05）. And the score of phlegm （dampness）-heat syndrome was lower than that in control group （P<0.01）， levels of FBG， PBG， HbA1c， HOMA-IR， AUCI30~I120/G30~G120， TC， TG， LDL-C， TNF-α， leptin and resistin were lower than those in control group （P<0.01）， while levels of I30/△G30， HOMA-β， InISI， HDL-C and adiponectin were higher than those in control group （P<0.01）. There was no adverse reaction related to modified Huanglian Wendantang.ConclusionIn addition to treatment with metformin， modified Huanglian Wendantang can effectively control blood glucose and lipid， regulate adipocyte factor， improve early and late phase insulin secretion， improve the function of β cell and insulin sensitivity of islet， improve IR， with a better comprehensive efficacy and a safety in clinical use.
Keywords：type 2 diabetes mellitus;newly diagnosed;Huanglian Wendantang;syndrome of phlegm （dampness） combined with heat;insulin resistance;islet β cells;adipocytokines
Abstract：ObjectiveTo discuss clinical effect of addition and subtraction therapy of Ditantang combined with Taohong Siwutang to cerebral infarction and syndrome of phlegm and blood stasis blocking collaterals during early recovery， and to study protection to brain nerve.MethodOne hundred and fifty-two patients were randomly divided into control group （76 cases） and observation group （76 cases） by random number table， 71 patients in control group completed the therapy （5 patients were falling off， missing visit or eliminated）， and 70 patients in observation group completed the therapy. Both groups' patients got comprehensive rehabilitation measures. Patients in control group got Zhongfeng Huichun pills， 1.5 g/time， 3 times/day. Patients in observation group got addition and subtraction therapy of Ditantang combined with Taohong Siwutang in the morning and at night， 1 dose/day. The treatment was continued for 12 weeks. Before and after treatment， scores of degree of neurological deficit， Barthel （BI） index， Fugl-Meyer scale （FMA）， modified Rankin scale （MRS） and syndrome of phlegm and blood stasis blocking collaterals were graded. And levels of malondialdehyde （MDA）， glutathione peroxidase （GSH-Px）， superoxide dismutase （SOD）， advanced oxidation protein products （AOPP）， vascular endothelial growth factor （VEGF）， brain-derived neurotrophic factor （BDNF） and neuron specific enolase （NSE）. And cerebral hemodynamics were detected， and peak flow velocity （VS）， vascular resistance index （RI）， pulsatility index （PI） and cerebrovascular reserve function （CVR） were recorded. Safety was evaluated.ResultAfter the 6th week and 12th week of treatment， scores of degree of neurological deficit， BI， FMA， MRS， syndrome of phlegm and blood stasis blocking collaterals， AOPP， MDA， NSE， RI and PI were lower than those in control group （P<0.01）， levels of SOD， GSH-Px， BDNF， VEGF， Vs and CVR were higher than those in control group （P<0.01）. The clinical effect was better than which in control group （Z=2.109， P<0.05）. Besides， there was no adverse reaction caused by Ditantang combined with Taohong Siwutang.ConclusionDitantang combined with Taohong Siwutang can ameliarate the hemodynamics， reduce the lipid peroxidation damage， regulate the neurovascular repair factor， so it can promote the repair of nerve tissue and function， clinically reduce the degree of nerve function defect， improve the ability of daily life and exercise when it used to cerebral infarction and syndrome of phlegm and blood stasis blocking collaterals during early recovery， and it is good for clinical effect and safe using.
Keywords：cerebral;syndrome of phlegm and blood stasis blocking collaterals;convalescence;Taohong Siwutang;Ditantang;cerebral hemodynamics;oxidative stress;nerve repair
Abstract：ObjectiveTo discuss clinical effect of Shexiang Tongxin pills to syndrome of Qi deficiency and blood stasis in ischemic heart failure （IHF） and to study the mechanism to myocardial fibrosis and vascular regeneration.MethodOne hundred and forty patients were randomly divided into observation group （70 cases） and control group （70 cases） with the same cases. 66 patients in control group finish the therapy （3 patients fall off or lost visit， 1 were eliminated）. 65 patients in observation group completed the therapy （4 patients were falling off， 1 were eliminated）. Both groups patients got combined treatment according to the guiding. Patients in control group got simulated medicine of Shexiang Tongxin pills， 2 pills/time， 3 times/day. Patients in observation group got Shexiang Tongxin pills， 2 pills/time， 3 times/day. The treatment was continued for 24 weeks. Before and after treatment， the left ventricular ejection fraction （LVEF）， cardiac output （CO）， end diastolic diameter （LVEDd）， left posterior wall thickness （LVPW）， left ventricular mass index （LVMI） and left ventricular remodeling index （LVRI） were measured by echocardiography. And scores of six minute walking test （6 MWT）， Lee's heart failure score， Qi deficiency and blood stasis syndrome score and Minnesota Heart Failure Quality of life questionnaire （MLHFQ） were graded. And levels of N-terminal B-type natriuretic peptide （NT-proBNP）， transforming growth factor-β1 （TGF-β1）， soluble ST2 （sST2）， matrix metalloproteinase-2 （MMP-2）， matrix metalloproteinase inhibitor-2 （TIMP-2）， insulin-like growth factor-1 （IGF-1）， vascular endothelial growth factor （VEGF）， angiopoietin1 （Ang1） and angiopoietin2 （Ang2）， and the safety was evaluated.ResultThe total clinical effect in observation group was 90.77% （59/65） higher than 74.24% （49/66） in control group （=6.179， P<0.05）. Levels of LVEF， CO， LVRI， 6 MWT， TIMP-2， IGF-1， VEGF and Ang1 were higher than those in control group （P<0.01）. And levels of LVEDd， LVPW and LVMI were lower than those in control group （P<0.05）， levels of NT-proBNP， TGF-β1， sST2， MMP-2 and Ang2 were lower than those in control group （P<0.01）， scores of Lee's heart failure score， Qi deficiency and blood stasis syndrome score and MLHFQ were lower than those in control group （P<0.01）. Besides， there was no adverse reactions caused by Shexiang Tongxin dropping pills.ConclusionOn the basis of conventional western medicine treatment， Shexiang Tongxin drop pills can improve the ventricular remodeling， improve the heart function， reduce the clinical symptoms， increase the exercise tolerance and quality of life of the patients by affecting the myocardial fibrosis and vascular regeneration factor， which has better clinical efficacy and safety.
Keywords：Qi deficiency and blood stasis in ischemic heart failure;Shexiang Tongxin pills;Qi deficiency and blood stasis;myocardial fibrosis;vascular regeneration;ventricular remodeling
Abstract：ObjectiveTo conduct quality evaluation of Ginkgo Folium preparations by analyzing the national evaluation sampling test results， analyze the quality differences， and put forward suggestions for the improvement of quality standards and market supervision.MethodThe contents of total flavonol glycosides and terpene lactones in Ginkgo Folium tablets and Ginkgo Folium capsules were determined according to the methods of determination in the 2015 edition of Chinese Pharmacopoeia （the first volume）， and the contents of free flavonoids （quercetin， kaempferide and isorhamnetin） and sophoricoside in Ginkgo Folium preparations were determined according to related supplementary testing method of Ginkgo Folium tablets and Ginkgo Folium capsules issued by National Medical Products Administration. The quality differences of Ginkgo Folium preparations from different batches and different manufacturers were compared according to the contents of total flavonol glycosides， terpene lactones， free flavonoids and sophoricoside in 328 batches of Ginkgo Folium tablets and Ginkgo Folium capsules manufactured by 48 enterprises.ResultQuality of 328 batches of Ginkgo Folium tablets and Ginkgo Folium capsules was in accordance with the standard， but the contents of terpene lactones and total flavonol glycosides were all distributed in a wide range， and the quality of samples varied greatly among different enterprises.ConclusionIt is recommended that each enterprise should optimize the production process and strictly control the raw materials to ensure the consistency between different batches of samples.
Abstract：ObjectiveTo investigate the relationship between phase behavior of curcumin （CUR） from self-nanoemulsion drug delivery system （SNEDDS） and stability of the formed nanoemulsion in artificial gastrointestinal fluid.MethodThe growth rate of precipitation after dispersion of CUR-SNEDDS was expressed by the change tendency of CUR supersaturation-time curve. The effect of drug loading on crystal nucleation and growth was investigated by ultraviolet-visible spectrometry and polarized light microscope， respectively. X-ray powder diffraction （XRD） and differential scanning calorimetry （DSC） were used to analyze the precipitation forms of CUR-SNEDDS with different drug loading in artificial gastrointestinal fluid. At the same time， the effect of drug loading on the quality stability of nanoemulsion formed by CUR-SNEDDS in artificial gastrointestinal fluid was investigated.ResultIn the artificial gastrointestinal fluid， with the increase of drug loading， the area under the supersaturation-time curve of CUR was increased （100% drug loading≈90% drug loading>75% drug loading）， the crystallization nucleation and growth rate were accelerated （100% drug loading>90% drug loading>75% drug loading）， the amorphous proportion in the precipitation composition decreased， the nanoemulsion droplets adhered and distributed unevenly， the particle size and dispersivity were increased.ConclusionHigh drug loading promotes the nucleation and growth of crystals， and increases the proportion of crystal forms in the precipitation composition， which leads to the decrease in the stability of the formed nanoemulsion. Therefore， it is suggested that the drug loading of CUR-SNEDDS needs to be controlled below 90%.
Abstract：ObjectiveTo establish the grade evaluation method for Codonopsis Radix slices based relative quality constants， in order to provide scientific theoretical basis for grading of Codonopsis Radix slices.MethodThrough literature and market research，the main production areas of Codonopsis Radix slices were determined，and 67 batches of Ludangshen slices（52 batches） and Baitiaodang slices （15 batches） were collected. The appearance traits （average quality and average thickness of Codonopsis Radix slices） were observed and measured. According to Chinese Pharmacopoeia （2015 edition）， the extract and the content of Codonopsis pilosula polysaccharide was determined by phenol-sulphoacid method. Then the relative quality constant was calculated，and the results of grade evaluation were evaluated through systematic cluster analysis and correlation analysis.ResultRelative quality constants of 67 batches of Codonopsis Radix slices were between 0.32-2.97. If these samples were divided into three grades：the first-grade relative quality constants were greater than or equal to 2.08，the second grade was greater than or equal to 0.89 but less than 2.08，while the third grade was less than 0.89. The results of systematic cluster analysis showed that 67 batches of Codonopsis Radix slices were clustered into 3 categories，and the results were basically consistent with the classification. The correlation analysis showed a positive correlation between the content of the extracts and the polysaccharide content （P<0.05）.ConclusionThis method links the extrinsic characteristics to the intrinsic quality，and objectively grade Codonopsis Radix slices， so as to provide a basis for its grade standards.
Abstract：ObjectiveTo study the resources and medicinal use of Tibetan medicine Codonopsis canescens，analyze its suitable habitat，and protect its wild resources，in order to provide an important reference basis for further rational development and utilization.MethodCombined investigation methods of literature textual research，visiting investigation and on-the-spot investigation were adopted.ResultIn this paper，we investigated the resources and medicinal use of C. canescens in 27 cities and counties of Sichuan，Qinghai and Tibet. Compared with historical records of C. canescens，the results showed that there was no obvious change in the distribution range，and the distribution was uneven in different places. Ganzi Prefecture， Sichuan， boasts rich resources； in particular， Luhuo county has as high as 90-120 plants/100 m2， it is estimated that the reserves per unit area of root C. canescens in this area were 3 445.32 g·m-2，which totaled 1 279.18 ton（1 ton=1 000 kg）. In history，the root is the main medicinal part. However， in recent years，in order to protect resources，the aboveground part is frequently used. The clinical efficacy of Tibetan medicine is mainly to tonify Qi and activate blood circulation. It can be combined with other drugs to treat rheumatism，sores，furuncle，carbuncle and other diseases. However，the plants of Codonopsis are mixed with C. canescens in each Tibetan area，and the mixed varieties are determined according to the growing varieties of the same genus in this area.ConclusionThe reserves of wild resources of C. canescens are decreasing with the increase of clinical use and market demand of Tibetan medicine. There is serious mixture of Codonopsis plants in Tibetan areas. It is suggested paying attention to resource protection，carrying out artificial planting，establishing the standard for the medicinal use of Tibetan medicine，and promote rational and sustainable utilization of resources.
Keywords：Tibetan medicine;Codonopsis canescens;resources investigation;current medicinal use
Abstract：ObjectiveTo study the effects of different drought conditions on the growth and photosynthetic physiological parameters of Acanthopanax senticosus，in order to provide the theoretical basis for standardized planting and rational development and utilization of A. senticosus.MethodIn this study，three-year-old A. senticosus was used as experimental samples. The growth parameters，photosynthetic parameters，and photosynthetic physiological parameters were determined to study the effects of different drought conditions on the growth and photosynthesis of A. senticosus.ResultThe plant height and leaf number were significantly lower than the control group under drought stress conditions，and the leaf area was higher than the control group under drought stress. Net photosynthetic rate，stomata conductance and transpiration rate were not significantly different between the control group and the moderate drought stress group. They were significantly decreased in the severe drought stress group，while the intercellular carbon dioxide concentration increased with the severity of drought stress. With the treatment time，the initial fluorescence was higher in the severe drought stress group than in the control group，and the moderate drought stress group was lower than the control group，the maximum fluorescence was significantly lower in the severe drought stress group than in the control group， potential photochemical efficiency and maximum photochemical efficiency were significantly elevated in the moderate drought stress group.ConclusionDrought stress can significantly inhibit the growth of A. senticosus. Severe drought conditions can significantly inhibit the photosynthesis of A. senticosus leaves. This effect is related to the regulation of stomatal size，but not related to the activity of photoreaction center.
Abstract：ObjectiveTo systematically evaluate the efficacy and safety of modified Buyang Huanwu Tang in the treatment of chronic heart failure.MethodCNKI database，Wanfang database，VIP database，Pubmed，MEDLINE，EMBASE and Cochrane database were retrieved systematically. The literature retrieval period is from no limit to December 2019，with "Buyang Huanwu Tang" and "chronic heart failure" "heart failure" as the key words for full-text retrieval of Chinese and English databases. Literatures of randomized controlled trials（RCTs） for chronic heart failure were included， and the data were extracted. Cochrane system evaluation method was used to score the quality of literature. Stata 14.0 was applied in Meta-analysis on the retrieval results. TSA0.9 was applied in test sequential analysis. Sensitivity analysis was made to explain heterogeneity，and funnel chart was used to evaluate publication bias.ResultA total of 2 037 patients were included in 21 RCT studies. The article quality risk assessment was generally unclear risk of bias. The results of meta-analysis showed that the left ventricular ejection fraction （LVEF） in the experimental group was significantly higher than that in the control group，with statistically significant differences ［MD=0.901，95% CI （0.772，1.029），P<0.01］，the left ventricular end diastolic diameter （LVEDd） in the experimental group was significantly lower than that in the control group，with statistically significant differences ［OR=-0.650，95% CI=（-0.854，-0.446），P<0.01］，BNP in the experimental group was significantly lower than that in the control group，with statistically significant differences ［MD=-1.212，95% CI=（-1.359，-1.066），P<0.01］，6-minute walk test （6MWT） in the experimental group was significantly higher than that in the control group，with statistically significant differences ［MD=0.797， 95% CI=（0.447，1.146），P<0.01］，and the effective rate in the experimental group was significantly improved，with statistically significant differences ［OR=1.840，95% CI=（1.680，2.016），P<0.01］.ConclusionModified Buyang Huanwu Tang combined with conventional western medicine treatment of chronic heart failure is more effective than single administration of western medicine treatment，and can improve clinical efficacy， effectively improve the LVEF of patients with chronic heart failure，reduce the LVEDd reduces plasma BNP levels，prolong the 6-minute walking distance，and reduce the incidence of adverse reactions.
Abstract：Based on the syndrome and treatment system of collateral disease， and inheriting the development of the bloodline theory of traditional Chinese medicine （TCM）， academician WU Yi-ling systematically constructed the vessel-collateral theory of TCM and proposed that its core theory was the theory of Yingwei， that is， "Ying in the vein， and Wei outside the vein" （Huangdi Neijing·Lingshu·Yingwei Shenghui）， "obstructing of Yingwei， congelation of blood" （Treatise on Febrile Diseases· Pulse Differentiation Method）， "pathogen transferring through blood vessels， obstructed by blocking" （Jinkui Yaolue·Zangfu Jingluo Xianhou Bingmaizheng First）， and "damage of the heart， adjust its Yingwei". Based on the consistency of vessel-collateral as the channel of blood circulation in TCM with the blood vessels in western medicine， and guided by the Yingwei theory of vessel-collateral theory， the diagnostic criteria of syndrome differentiation of disease of vessel-collateral and vascular system represented by coronary heart disease， arrhythmia， heart failure and others was established to guide the prevention and treatment of vascular diseases. Based on the above analysis， guided by the Yingwei theory of vessel-collateral theory， and on the basis of related researches of vessel-collateral and vascular system， this paper discussed the etiology and pathogenesis of chronic coronary syndrome （CCS） in TCM. Taking useful collateral with unblocking as the treatment principle， the representative Tongluo prescription （Tongxinluo） was constructed， the research progress of Tongxinluo from various aspects such as animal experiment research， pharmacological research and clinical evidence-based research was summarized， a comprehensive system from etiology， pathogenesis， syndrome differentiation to treatment was formed， in order to provide new ideas for the clinical treatment of CCS.
Keywords：vessel-collateral doctrine;Yingwei theory;disease of vessel-collateral and vascular system;chronic coronary syndrome;Tongxinluo
Abstract：ObjectiveThe incidence rate of cancer cachexia is high in late stage of cancer， which is characterized by skeletal muscle atrophy and rapid reduction of adipose tissue and systemic inflammation. Cachexia is highly related to a variety of tumors， and causes a large proportion of cancer deaths. Cancer cachexia can lead to serious complications in patients with cancer， then， the quality of life of patients decreases， the psychological state becomes negative， and the state of illness is further worsened. At present， there is no effective intervention means to completely reverse cachexia. The combined use of multiple targets and effective components of traditional Chinese medicine（TCM）， as well as the treatment based on syndrome differentiation and the theory of TCM play an important role in the prevention and treatment of cancer cachexia. Therefore， exploring the pathogenesis of cancer cachexia and prevention and treatment with TCM is helpful for basic study and clinical application.MethodIn this paper， cancer cachexia and TCM in China national knowledge infrastructure （CNKI） and Public Medline （PubMed） databases were retrieved， and 98 Chinese and English literatures were included through summarization to elaborate the pathogenesis of cancer cachexia and the prevention and treatment of TCM.ResultEmphasis was given to the important role of inflammatory reaction， skeletal muscle atrophy， energy metabolism abnormality and multiple signal joint regulation in occurrence of cancer cachexia， and the unique advantages and significant role of TCM in treatment of cancer cachexia under different treatment principles.ConclusionInflammatory reaction， skeletal muscle atrophy， abnormal energy metabolism make the pathogenesis of cancer cachexia complex and diverse. TCM prescriptions， Chinese herbal medicine and their effective ingredients have the natural advantages of targeting multiple pathways， controlling multiple signal pathways and inhibiting various inflammatory factors in the prevention and treatment of cancer cachexia， and are safe and effective in improving diet， prolonging the survival period of patients and keeping weight.
Keywords：cancer cachexia;skeletal muscle atrophy;traditional Chinese medicine;prescription
Abstract：Medicinal plant germplasm resources are the foundation of the modern development of traditional Chinese medicine. In-depth study of medicinal plant germplasm resources is a prerequisite for cultivating fine varieties and ensuring the output and standard quality of traditional Chinese medicine（TCM）. Traditional identification methods start with appearance and are greatly affected by natural environment and human factors，with a low efficiency and accuracy of identification are generally low molecularin general. Due to such advantages as easy operation，high sensitivity，accurate results， molecular biology technology has been widely used in the related research of relevant studies for medicinal plant germplasm resources due to its advantages of easy operation，high sensitivity，accurate results，etc. It mainly involving the distinction between wild and cultivated products，researchstudy on substitutes of TCM，identification of Chinese patent medicine，good variety marker breeding，genetic diversity researchstudy，genetic map establishment and omics research，etcstudy. Among them，omics researchstudy is divided into genomics，transcriptomics，metabolomics，and proteomics due toby different analysis purposes. Genomics is divided into three sub-fields namely structural genomics，functional genomics， and comparative genomics. Eukaryotes Because eukaryotes have nuclei and organelles，so omics researchstudy also includes chloroplast genomics，mitochondrial genomics，nuclear genomics，and plastid genomics. Among them，the chloroplast genome has a simple structure，small molecular weight，and good conservation，while the mitochondrial genome has a strong variability and complex structure，the nuclear genome data isfeatures complex， data and the nucleus contains no ribosomes in nucleus，resulting in spatiotemporal differences in the translation process，even if repeated repeatedly test， the result of and the test is alsoresults remained uncertain， even after repeated tests. The molecular biology technology and omics researchstudy involved in theby current medicinal plant researchstudy still hashave shortcomings，and there iswith a large room for development，which needs and need further improvement and supplementation. This articlepaper successively introduces the characteristics and applications of cytology，molecular markers，and omics researchstudy techniques in the identification of medicinal germplasm resources，providingin order to provide a reference for subsequent identification，development and utilization of medicinal plant germplasm resources.
Keywords：medicinal plants;cytology study;DNA molecular markers;omics study
Abstract：Mineral medicine is an indispensable part of traditional Chinese medicine and has a long history of application. Among them， mineral-based hemostatics have been widely applied for the treatment of various hemorrhagic diseases with extensive clinical experience and established efficacy. Gypsum Fibrosum （GF）， a commonly used mineral medicine in clinical， can clear away heat， and relieve anxiety and thirst. Gypsum Ustum （GU） is the processed product of GF after calcining at high temperature. It is mainly composed of anhydrous calcium sulfate （CaSO4） with the functions of moisturizing， promoting muscle growth， astringent sores and hemostasis. GU is often used externally to treat ulcer， itching， eczema， water and fire scalds， trauma bleeding， etc. Studies on the mechanism of hemostasis have shown that Ca2+ （coagulation factor Ⅳ） is involved in many key processes of the internal and external coagulation cascades and can prevent bleeding by regulating platelet activation and aggregation， and promoting the production of insoluble fibrin and the ultimate formation of a blood clot. GF and GU both contain Ca2+ which provide an important material basis of hemostatic effect for both compounds， but GU has a significant hemostatic effect， while GF has no hemostatic effect. After processing， the taste and efficacy of the GF have been obviously changed which reflects the characteristics of processing， but the processing mechanism of GU has not been fully clarified. Therefore， based on studies of GF before and after calcining， this paper focused on these aspects including calcining process， crystal form comparison， element content， efficacy comparison， and summarized various aspects of Ca2+ involved in hemostasis. In addition， the hemostatic properties of other calcium-containing mineral medicines and new calcium-containing hemostatic materials such as calcium alginate， mesoporous calcium silicate and nanogel hemostatic materials were also discussed. The paper aimed to provide a reference for elucidating processing mechanism and clinical dialectical use of GU， also to promote development of new calcium-containing hemostatic materials.
Keywords：Gypsum Ustum;calcium ion （Ca2+）;mineral medicine;material basis;processing mechanism;coagulation pathway;hemostasis
Abstract：Biliary tract cancers are group of highly heterogeneous malignant tumors derived from gallbladder and bile duct epithelium， including gallbladder cancer and cholangiocarcinoma， with a high degree of malignancy. When diagnosed， the course of disease often develops to the middle and late stage， with a poor prognosis. It is difficult to treat the advanced biliary tract cancers， with a lack of unified chemotherapy standard and targeted drugs. Traditional Chinese medicine（TCM） compound has multi-target， multi-link and multi-path characteristics， with effects in stabilizing tumor， alleviating clinical symptoms， alleviating side effect of radiotherapy and chemotherapy， and improving the quality of life. Especially in the treatment of advanced cancer patients， it plays an irreplaceable role. Therefore， it is concerned by many researchers. According to TCM， the pathogenesis of patients with advanced biliary tract cancers are mainly due to asthenia of healthy Qi and sthenia of pathogenic factors， Yin-Yang disharmony， and simultaneous occurrence of cold and heat. The location of the disease is related to liver， gallbladder， spleen， stomach and kidney. Because the advanced biliary tract cancers are similar to Jueyin syndrome in disease location， course and disease， Wumeiwan is often used in clinical treatment. Based on "theory， method， prescription and medicine" of TCM， Wumeiwan has the effect in regulating cold and heat， clearing the upper and lower temperature， warming the liver and supporting Yang， attacking and supplementing simultaneously， balancing the liver， spleen and kidney， which conforms to the TCM syndrome characteristics of advanced biliary tract cancers， according to modern Chinese medicine pharmacology， many effective ingredients in Wumeiwan have the effects in inducing tumor cell apoptosis， inhibiting tumor cell proliferation and reducing tumor cell proliferation， reducing inflammatory response in the body， inhibiting angiogenesis， reversing multidrug resistance of tumor cells and regulating immunity. This paper analyzes the feasibility of treatment of advanced biliary tract cancers with Wumeiwan from the perspective of traditional Chinese and Western medicine， so as to provide reference for clinical comprehensive treatment of advanced biliary tract cancers and the experimental study of TCM.