Abstract：ObjectiveTo observe the effect of Dabuyuan Jian on the synaptic plasticity of hippocampus and the brain derived neurotrophic factor （BDNF）/tyrosine kinase receptor （TrkB）/cyclic adenosine phosphate reactive element binding protein （CREB） signaling pathway in amyloid precursor protein/presenilil （APP/PS1） mice， and to explore its possible mechanism for improving synaptic plasticity.MethodTotally 36 APP/PS1 mice were divided into model group， donepezil group（6.5×10-4 g·kg-1·d-1） and Dabuyuan Jian group （13.2 g·kg-1·d-1）， and another wild mice were set as control group. The mice in control group and model group received an equal volume of saline， and the mice in each group received drugs by gavage for 30 days. The learning ability and memory of mice in each group were detected by Morris water maze. The pathological changes of neurons and synapses in the hippocampus of each group were observed by Nissl staining and Golgi staining. The expression levels of postsynaptic density protein 95 （PSD95） and synaptophysin （SYN） in hippocampus of each group were detected by immunofluorescence （IF）. The protein expression levels of BDNF， TrkB， CREB and phosphorylated CREB （p-CREB） in hippocampus were detected by Western blot.ResultAs compared with the control group， the platform latency and total swimming distance of the model group were increased in the model group （P<0.01）， with decreased times of crossing the platform and staying time in the target quadrant （P<0.01）， the intracellular Nissl bodies of neurons in hippocampal CA3 area decreased or disappeared in model group， with decreased number of neurons and dendritic branches and decreased density of dendritic spine in hippocampal CA3 area of the mice （P<0.01）， and the protein expression levels of SYN， PSD95， BDNF， TrkB and p-CREB in hippocampus of mice were also decreased in model group （P<0.05， P<0.01）. As compared with the model group， the platform latency and total swimming distance were decreased in the donepezil group and Dabuyuan Jian group （P<0.05， P<0.01）， with increased times of crossing platform and staying time in target quadrant （P<0.05， P<0.01）， the number of Nissl bodies of neurons in hippocampal CA3 area was increased in the donepezil group and Dabuyuan Jian group， with increased number of neurons and dendritic branches and increased density of dendritic spine in hippocampal CA3 area of the mice， and the protein expression levels of SYN， PSD95， BDNF， TrkB and p-CREB in hippocampus of mice were increased in the donepezil group and Dabuyuan Jian group （P<0.05， P<0.01）.ConclusionDabuyuan Jian can improve the synaptic plasticity of APP/PS1 double transgenic mice， and its mechanism may be related to its up-regulation of BDNF/TrkB/CREB signal pathway in mouse hippocampus.
Abstract：ObjectiveThrough the preparation of Alzheimer's disease （AD） rat model， the effect of Yuanzhisan on the expression of Ghrelin was observed， and the possible mechanisms in preventing and treating AD were discussed.MethodA total of 120 SD rats were randomly divided into sham-operated group， model group， donepezil group（1.02 mg·kg-1）， and high， medium and low-dose Yuanzhisan groups （12，6，3 g·kg-1）， with 20 rats in each group， including half male and half female. The rats in sham-operated group were injected with normal saline （NS）， and the rats in other groups were injected with β-amyloid 1-40 （Aβ1-40） in hippocampus to induce the AD rat model. During the 10-week continuous gavage， the food intake of rats in each group was observed and recorded. After the end of gavage， learning and memory abilities of rats were tested by Morris water maze. The whole brain and the gastric body and antrum were collected， the pathologic changes in the CA1 area of hippocampus was assessed by hematoxylin-eosin （HE） staining， and the expression of Ghrelin was detected by immunohistochemistry.ResultCompared with the sham-operated group， the escape latency time of model group rats increased （P<0.01），while times across platform， retention time in effective area and movement distance decreased （P<0.01）. The disorder of neurons， the decrease of the neuronal number， and the pyknosis of nucleus were observed in hippocampal CA1 area. The food intake of male and female rats decreased significantly （P<0.05，P<0.01）. The expression of Ghrelin in hippocampal CA1 area and gastric mucosa decreased significantly （P<0.01）. Compared with the model group， the escape latency time of rats in each treatment group was significantly shortened （P<0.05，P<0.01），whereas times across platform， retention time in effective area and movement distance increased （P<0.05，P<0.01）. The pathologic change was improved markedly， and the daily food intake of rats in high and medium-dose Yuanzhisan groups increased significantly （P<0.05，P<0.01）. The protein expression of Ghrelin in hippocampal CA1 area and gastric mucosa increased significantly in each Yuanzhisan group （P<0.05，P<0.01）.ConclusionYuanzhisan can effectively improve the learning and memory abilities of AD rats and increase the daily intake， which may be related to its up-regulation of Ghrelin content in hippocampal CA1 area and gastric mucosa.
Abstract：ObjectiveTo investigate the effects of Buyang Huanwu Tang （BHT） on axonal regeneration and neurological rehabilitation of the rats suffering ischemic stroke （IS）.MethodA total of 180 SD rats were used to establish a middle cerebral artery infarction （MCAO） model. The animals that were successfully modeled were randomly divided into model group， BHT group （12 g·kg-1） and nimodipine group （20 mg·kg-1）， and a sham group was established， with 28 rats in each group. After seven-days intragastric administration of BHT， the animals were sacrificed. TTC staining was used to test cerebral infarction. Brain water content was measured to observe cerebral edema. Bielschowsky's silver staining and immunofluorescence were performed to observe axonal degeneration and the protein expression of neurofilament protein-200（NF-200）. Quantitative real-time polymerase chain reaction （PCR） was used to analyze the mRNA expression of repulsion oriented molecule a （RGMa）， Ras homologous enzyme （Rho）， Rho kinase （ROCK）， and collapsion response regulatory protein 2 （CRMP2）. Neurological function scores assay was used to examine neurological recovery.ResultCompared with sham group， the cerebral infarction volume and brain water content increased significantly（P<0.01）， and motor function was markablely decreased in the model group. Axonal degeneration and nerve fiber damage were obviously observed. Also， gene expression of axon growth-related protein was deviation from normal （P<0.01）. Compared with model group， the cerebral infarction rate （P<0.01）， brain water content （P<0.01） and axonal degeneration of BHT group and nimodipine group were significantly reduced. The expression of NF-200 was increased. Also， the mRNA expression of RGMa， Rho and ROCK was lower （P<0.05） while the mRNA expression of CRMP2 was higher （P<0.01）. And the neurological function was significantly improved （P<0.05）.ConclusionBHT can promote axon regeneration after ischemic stroke injury by regulating the mRNA expression of axon growth-related protein， thereby improving nerve function.
Abstract：ObjectiveTo observe the changes of leptin receptor-tyrosine kinase Janus2 （JAK2）/signal transducer and activator of transcription 3 （STAT3） pathway and the regulatory effect of Xiaoyaosan on the hypothalamic arcuate nuclei of rats with chronic mild unpredictable stress model （CUMS）.MethodSixty male sprague-dawley rats were randomly divided into normal group， model group， Xiaoyaosan group， and fluoxetine group. After one-week adaptive feeding， the rats in model group， Xiaoyaosan group and fluoxetine group were uesd to replicate the chronic psychological stress rat model through mild unpredictable stimulation. Meanwhile ， they were simultaneously administered the corresponding drugs， Xiaoyaosan 19.27 g·kg-1·d-1， Fluoxetine 2 mg·kg-1·d-1 （based on the average adult body weight of 60 kg）， the rats in the normal group and the model group were given the same volume of normal saline for 6 weeks. The body weight， food intake， sucrose consumption ratios， and the experimental behavior in the open field test （OFT） of the groups were observed. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to detect the expressions of ob-R， JAK2， and STAT3 in the arcuate nucleus of rat hypothalamus.ResultCompared with the normal group， the body weight and food intake of the model group were significantly decreased （P<0.05， P<0.01）， the sucrose consumption ratios ， the total behavioral distance of the experimental field and the total distance of the central area were significantly reduced， the protein and mRNA expressions of ob-R， JAK2， STAT3 in the arcuate nucleus of hypothalamus in rats increased significantly （P<0.05， P<0.01）. Compared with the model group， the body weight of Xiaoyaosan group increased significantly on the 7th， 14th， 21st and 28th days （P<0.05， P<0.01）， the food intake of rats increased significantly on the 21st and 35th days of the experiment （P<0.05）， and the sucrose consumption ratios， the total distance of the experimental behavior in the open field test （OFT） and the total distance of the central area were significantly improved. Xiaoyaosan had a corresponding regulatory effect on the protein and mRNA expressions of ob-R， JAK2， STAT3 in the arcuate nucleus of hypothalamus in model rats （P<0.05， P<0.01）.ConclusionXiaoyaosan regulates the body weight， appetite， and energy metabolism of chronically mild and unpredictable stress rats， which may be related to the ob-R-JAK2/STAT3 pathway in the hypothalamic arcuate nucleus.
Abstract：ObjectiveThe present study was to observe the effect of Qianjin Weijingtang on the differentiation of helper T cells 17 （Th17）/T regulatory cell （Treg） and the expressions of related cytokines in the lung tissues of the model rats exposed to cigarette smoke.MethodTotolly 60 male rats were randomly assigned into six groups （control group， model group， acetylcysteine group and Qianjin Weijingtang high， moddle and low dose groups）， with 10 rats in each group. After 30 day's modeling and 30 day's intervention， rats were killed peacefully with their tissues collected. Real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） was used to detect the expression of retinoic acid associated orphan receptor （ROR-γt） and forkhead/pterygoid helix transcription factor 3 （Foxp3） mRNA， enzyme-linked immunosorbent assay （ELISA） was used to check the concentration of interleukin-17（IL-17）， IL-6， IL-10 and transforming growth factor-β1（TGF-β1）. Htoxylin eosin （HE） staining was used to observe the pathological changes of lung tissues， while flow cytometry was used to detect Treg（Foxp3+CD25+）， Th17（CD4+IL-17+） and Treg/Th17 ratio.ResultAs compared with the control group， the ROR-γt mRNA expression in model group was higher（P<0.01）. As compared with the model group， various doses of Qianjin Weijingtang down-regulated the expression（P<0.05，P<0.01）. As compared with the control group， the expression of the Foxp3 mRNA was down-regulated in model group（P<0.05）， but was up-regulated in Qianjin Weijingtang middle and low dose groups（P<0.05）. Compared with the control group， the concentration of IL-17 and IL-6 in the model group were significantly increased， while the concentration of IL-10 and TGF-β1 in the model group were significantly decreased（P<0.05，P<0.01）. As compared with the model group， intervention with various doses of Qianjin Weijingtang could help to decrease the concentration of IL-17 and IL-6 in lung tissues， and increase the concentration of IL-10（P<0.05）， which were consistent with those of Real-time PCR results. Flow cytometry examination showed that the Th17（CD4+IL-17+） proportion decreased and Treg/Th17 ratio increased after Qianjin Weijing Tang middle and low dose intervention（P<0.05，P<0.01）.ConclusionQianjin Weijingtang could regulate Treg/Th17 ratio and help to achieve Th17/Treg balance.
Keywords：Qianjin Weijingtang;cigarette smoking;helper T cells 17 （Th17）;T regulatory cell （Treg）
Abstract：ObjectiveTo study the effect of Wuzi Yanzong Wan on the expressions of voltage-dependent anion channel 1 （VDAC1）， adenine nucleotide transposase （ANT）， cyclophilin D （CypD） and other proteins， and analyze its mechanism in intervening with sperm mitochondrial permeability transition pore （mPTP） opening.MethodForty rats were randomly divided into 4 groups， namely normal group， model group， positive group （Shengjing capsule， 1.6 g·kg-1·d-1）， and Wuzi Yanzong Wan group （4.0 g·kg-1·d-1）， with 10 rats in each group. Except for the normal group， tripterygium wilfordii glycosides （GTW， 30 mg·kg-1） was intragastrically administered for 8 weeks to establish the oligozoospermia model. After the 4th week， each group was given drugs through intragastric administration for 4 weeks， and fasted for 12 h after the last administration. These rats were anesthetized with 3% chloral hydrate， and their testis and epididymis tissues were collected. Western blot was used to determine the protein expressions of VDAC1，ANT，CypD，B-cell lymphoma/leukemia2 （Bcl-2）， Bcl-2 associated X protein （Bax）， Caspase-3， Caspase-9 in rat testis， Testicular tissue and its ultrastructure were observed under electron microscopy. The apoptosis in spermatogenic cells was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling （TUNEL）.ResultWestern blot results showed that compared with the normal group， the expressions of VDAC1， CypD， Caspase-3， Caspase-9 and Bax/Bcl-2 in the model group were increased （P<0.01）. The expressions of VDAC1， CypD， Caspase-3， Caspase-9 and Bax/Bcl-2 were significantly decreased in the positive group and the Wuzi Yanzong Wan group compared with the model group （P<0.01）. There was no significant change in the expressions of ANT and Bcl-2 protein between the groups. Testicular ultrastructural evaluation showed different sizes and disordered arrangement of sperm mitochondria and a large number of swelling and vacuoles in the model group， while complete structure and neat arrangement of sperm mitochondria and much less swelling and vacuole in positive group and Wuzi Yanzong Wan group. TUNEL results showed that the apoptosis rate of spermatogenic cells in the model group was significantly higher than that of the normal group （P<0.01）， while the apoptosis rate in the positive drug and Wuzi Yanzong Wan group was significantly lower than that of the model group （P<0.01）.ConclusionWuzi Yanzong Wan may resist germ cell apoptosis by inhibiting the expressions of VDAC1， CypD and Bax， reducing the permeability of mPTP， and preventing the cascade activation reaction of the Caspase family of apoptosis proteins.
Abstract：ObjectiveTo investigate the protective effect of Wutou Chishizhi Wan on myocardial ischemia reperfusion injury （MIRI） in rats， and observe its effect on such mechanisms as coagulation function， vascular endothelial cells and oxidative stress in rats.MethodA total of 40 SD rats were randomly divided into normal group， model group， positive drug group （Urokinase group） and Wutou Chishizhi Wan group， with 10 rats in each group. Except for the normal group， rat myocardial ischemia-reperfusion injury models were established. The changes of heart rate （HR） at 10 min before ischemia， 30 min after ischemia and 30， 60， 120 min （T0，T1，T2，T3，T4）， and the change of electrocardiogram （ECG） J point after modeling in rats were observed. The pathological changes of rat myocardial tissue were observed by hematoxylin-eosin （HE） staining. The changes of four indexes of coagulation ［prothrombin time （PT）， activated partial thromboplastin time （APTT）， thrombin time （TT）， fibrinogen content decreased significantly （FIB）］ in rats were observed. The contents of endothelin-1 （ET-1）， thromboxane A2 （TXA2） and prostacyclin （PGI2） in serum and myocardium levels of superoxide dismutase （SOD）， malondialdehyde （MDA） and glutathione peroxidase （GSH-Px） of MIRI rats were observed. Western blot assay was used for the detection of oxidative stress protein Keap1 and transcription factor-E2-related factor （Nrf2） expression levels in rat myocardial tissue.ResultCompared with the normal group， the ECG of MIRI rats showed significant myocardial ischemic injury-like changes， ST segment was significantly elevated， J point was significantly increased， and the incidences of HR in T1， T2， T3 and T4 were significantly reduced （P<0.05， P<0.01）. Compared with the model group， Wutou Chishizhi Wan significantly reduced ECG J-point changes in MIRI rats， while increased the incidence of HR in T1， T2， T3 and T4 （P<0.05， P<0.01）. Compared with the normal group， PT， APTT and TT in the model group were significantly shortened （P<0.01）， FIB content was significantly increased （P<0.01）， and the serum PGI2 level decreased and TXA2 and ET-1 levels increased significantly in the model group （P<0.01）. SOD content and GSH-Px activities of myocardial tissue in the model group were significantly reduced （P<0.01）， whereas the MDA content was increased （P<0.01）. Compared with the model group， PT of the Wutou Chishizhi Wan group was prolonged （P<0.05） and APTT slightly prolonged， TT significantly prolonged （P<0.01）， FIB content decreased （P<0.05）， serum PGI2 increased （P<0.05）， TXA2 and ET-1 decreased significantly in the Wutou Chishizhi Wan group （P<0.01）， myocardial MDA content decreased， and SOD content and GSP-Px activity increased significantly （P<0.01）. Meanwhile， the Wutou Chishizhi Wan group was able to activate the Keap1/Nrf2 signaling pathway， which significantly increased Nrf2 expression and significantly decreased Keap1 expression （P<0.01）.ConclusionWutou Chishizhi Wan group can protect myocardial injury in MIRI rats. The specific mechanism is to protect MIRI by regulating vascular endothelial cell homeostasis and oxidative stress levels and activating Keap1/Nrf2 signaling pathway.
Abstract：ObjectiveTo observe the influence of Chang'an Ⅰ prescription drug-containing serum on IgE-mediated RBL-2H3 cell degranulation model， and explore the mechanism of Chang'an Ⅰ prescription in inhibiting RBL-2H3 activation degranulation and releasing inflammatory mediators with v-yes-1 Yanaguchi sarcoma viral related oncogene homolog （Lyn）/spleen tyrosine protein kinase （Syk）/mitogen-activated protein kinase （MAPK） signal pathway.MethodPreparation for Chang'an Ⅰ prescription serum. Animal group， SD male rats were randomly divided into Chang'an Ⅰ prescription serum high， medium， low dose， and blank control groups with 10 rats in each group. Dosage： 10 mL·kg-1 distilled water was given to blank control group， while Chang'an Ⅰ prescription serum high， medium and low dose groups were respectively given to the Chang'an Ⅰ prescription concentrated crude drug with concentration of 1.15，2.30，4.60 g·kg-1， respectively once a day for 7 days continuously and then blood was taken from aorta ventralis and centrifuged. Ketotifen as the positive control drug. Mast cells are counted with toluidine blue staining. Cellular release of β-aminohexose was detected by colorimetric method. Contents of MCT， TNF-α， MCP-1 and histamine were measured by enzyme-linked immunosorbent assay （ELISA） kits， Lyn/Syk/MAPK protein levels were detected by immunoblotting.ResultFor cell activation and degranulation， compared with the blank control group， the model group had more cell degranulation （P<0.05）， compared with model group， the cell degranulation rate of each dose group of Chang'an Ⅰ prescription decreased （P<0.05）. The release rate of β-hexosamine in each dose group of Chang'an Ⅰ prescription decreased significantly （P<0.01）. For the release of active mediators， compared with the blank control group， the contents of histamine， MCT， TNF-α and MCP-1 all increased in the model group （P<0.01）， compared with the model group， the contents in each dose group of Chang'an Ⅰ prescription all decreased significantly （P<0.01）. Compared with the normal group， the phosphorylation levels of Lyn and Syk， extracellular regulatory protein kinase 1/2（ERK1/2）， c-Jun N-terminal kinase （JNK）， and mitogen-activated protein kinase p38 increased in the model group （P<0.05）. Compared with the model group， the Lyn， Syk and ERK1/2， JNK and p38 protein phosphorylation levels reduced in Chang'an Ⅰ prescription group （P<0.05）.ConclusionChang'an Ⅰ prescription drug-containing serum down-regulates the phosphorylation levels of proteins Lyn， Syk， and ERK1/2， JNK， and p38， inhibits RBL-2H3 cell activation and degranulation， reduces the release of cytokines and chemokines， such as histamine， MCT， TNF-α and MCP-1， it may be one of its mechanisms for treating IBS-D visceral hypersensitivity.
Abstract：ObjectiveTo investigate the effect of Tianwang Buxin pills on behavior， hypothalamus pituitary adrenal axis （HPA axis）， hippocampal glycogen synthase kinase 3β （GSK3β） phosphorylation and brain-derived neurotrophic factor （BDNF） expression in mice with chronic unpredictable stress， and explore its mechanisms for antidepressant-like action.MethodTotally 60 male ICR mice were randomly divided into normal group， chronic stress model group， fluoxetine hydrochloride group （10 mg·kg-1） and Tianwang Buxin pills high， middle and low dose groups （3.6， 1.8， 0.9 g·kg-1）. The mice were subjected to the chronic unpredictable stress （CUS） protocol for a period of 28 d to induce depressive-like behavior. Then， a sucrose preference test， open-field test and novelty-suppressed feeding test were performed to detect the behavior changes. The blood， adrenal gland and hippocampus of mice were collected. The contents of adrenocorticotropic hormone （ACTH） and corticosterone （CORT） in serum were detected by enzyme-linked immunosorbent assay （ELISA）. The changes of GSK3β phosphorylation and BDNF expression in hippocampus were detected by Western blot， and the adrenal index was then calculated.ResultAs compared with the normal group， the sucrose water preference was significantly decreased （P<0.01）， the number of opening activities was significantly reduced （P<0.05）， the feeding latency of novelty inhibition was prolonged （P<0.01）， the serum ACTH and CORT contents were significantly increased （P<0.05，P<0.01）， GSK3β phosphorylation and BDNF expression levels in hippocampus were significantly decreased （P<0.01）， and adrenal index was significantly increased in model group （P<0.01）. As compared with the model group， Tianwang Buxin pills treatment significantly reversed the CUS-induced behavioral abnormalities in depression model mice （P<0.05， P<0.01）， significantly decreased the levels of plasma ACTH and CORT （P<0.01） and adrenal and adrenal gland index （P<0.01）， while increased GSK3β phosphorylation and BDNF expression in hippocampus （P<0.05， P<0.01）， with its effect similar to that of fluoxetine hydrochloride.ConclusionTianwang Buxin pills produced antidepressant-like effects in chronic unpredictable stress mice， and its mechanism may be associated with inhibiting HPA axis activity and up-regulating GSK3β phosphorylation and BDNF protein expression in hippocampus.
Abstract：ObjectiveTo observe the effect of Huayu Jiedu recipe （HYJDR） on apoptosis of gastric cancer cells， explore the mechanism of HYJDR on apoptosis of gastric cancer cells and provide experimental basis for clinical application of HYJDR.MethodThiazolyl blue tetrazolium bromide （MTT） method was used to detect the effect of HYJDR （5，10，15，20，25，30 g·L-1） on the activity of SGC-7901 cells and the median inhibition concentration （IC50） of HYJDR on SGC-7901 cells. Propidium iodide （PI） and Hoechst double fluorescence staining were used to detect the effect of HYJDR on the apoptosis of gastric cancer SGC-7901 cells. Western blot was used to detect the effect of HYJDR on the expression of apoptosis related proteins in gastric cancer cells. Real-time polymerase chain reaction （Real-time PCR） was used to detect the effect of HYJDR on the expression of apoptosis related protein mRNA in gastric cancer cells.ResultAs compared with blank group， HYJDR （>10 g·L-1） can significantly inhibit the proliferation of gastric cancer cells. With the increase of the concentration of HYJDR， the cell viability decreased significantly obviously in a concentration-dependent manner（P<0.05）. HYJDR can significantly promote the apoptosis of gastric cancer cells. With the increase of the concentration of HYJDR， the apoptosis of gastric cancer cells gradually increased. As compared with blank group， HYJDR can obviously inhibit the expression of apoptosis-related B -cell lymphoma-2 （Bcl-2） （P<0.05）， and increase the expression of apoptosis-promoting proteins such as Bcl-2 antagonist of cell death （Bad） and Bcl-2-associated X （Bax）（P<0.05）. Real-time PCR results showed that as compared with blank group， HYJDR（5，10 g·L-1） could effectively inhibit the expression of Bcl-2 protein mRNA in gastric cancer cells（P<0.05）， and all HYJDR groups could promote the expression level of Bad mRNA（P<0.05）.ConclusionsHYJDR can obviously promote the apoptosis of SGC-7901， and its effect is probably achieved by increasing the expression of Bad mRNA and inhibiting the expression of Bcl-2 protein.
Abstract：ObjectiveTo explore the therapeutic effect and mechanism of Chaihu Yueju decoction on model rats induced by chronic unpredictable mild stress （CUMS）.MethodThe 60 SD rats were randomly divided into normal group， model group， Chaihu Yueju decoction low， medium and high-dose groups（0.3， 0.6， 1.2 g·kg-1） and fluoxetine hydrochloride group（0.2 mg·kg-1）， 10 for each group. The model rats was established by chronic unpredictable mild stress for 5 weeks， and then Chaihu Yueju decoction and fluoxetine hydrochloride were given to the corresponding treatment group by gavage for 3 weeks. In the last week of gavage， Morris water maze training and testing were conducted. After the last day of gavage， sugar water preference and other behavioral experiment were tested. The sugar water preference test was used to detect the degree of pleasure deficiency in rats before and after treatment， the open field test was used to detect the depression of rats before and after treatment， the spatial memory ability was tested by Morris water maze. Western blot was used to detect the levels of tumor necrosis factor-α（TNF-α）， 5-hydroxytryptamine receptor 1A （5-HT1A）， extracellular signal-regulated kinase （ERK） and synapse associated protein in hippocampus of each group， Gloji and Nissl staining was used to observe the changes of dendritic spines and Nissl bodies in the hippocampus CA3.ResultCompared with normal group， the weight， sugar water preference rate， the scores of horizontal and vertical movement were significantly reduced （P<0.05）， and the immobility time was significantly prolonged in model group （P<0.05）. Meanwhile， the ability of learning and memory in model group decreased （P<0.05）. Western blot results showed that the levels of TNF-α was significantly increased （P<0.05）， and the levels of 5-HT1A， p-ERK， cyclic adenosine phosphate reactive element binding protein （CREB）， p-CREB， Synapsin-1，Synaptophysin， glutamate receptor subtype-1（GluR-1）and postsynaptic membrane protein-95（PSD-95） in model group were significantly decreased （P<0.05）. Gloji and Nissl staining results showed that the density of dendritic spines and the number of Nissl body in the hippocampal CA3 of the model group was obviously attenuate （P<0.05）. The results show that the neurons were evidently damaged. Compared with model group， the weight， sugar water preference rate， the scores of horizontal and vertical movement were clearly increased （P<0.05） and the immobility time was significantly shortened in fluoxetine hydrochloride group， middle dose and high dose Chaihu Yueju decoction group （P<0.05）. Western blot results showed that the levels of synaptophysin， GluR-1 and PSD-95 were significantly increased in fluoxetine hydrochloride group， middle dose and high dose Chaihu Yueju decoction group. The level of TNF-α was significantly decreased （P<0.05）， the levels of 5-HT1A， p-ERK， CREB， p-CREB and Synapsin-1 were remarkably increased （P<0.05）， especially the high dose group of Chaihu Yueju decoction. Gloji and Nissl staining results showed that the density of dendritic spines and the number of Nissl body in the hippocampal CA3 of high dose group were similarly increased in a dose-dependent manner.ConclusionChaihu Yueju decoction could improves the weight， the depressive despair， autonomous activity ability and learning and memory ability of DP model rats. Its mechanism is closely related to attenuating the inflammatory reaction and enhancing the levels of 5-HT1A receptor protein， ERK and synapse related protein， then activating 5-HT/CREB and ERK/CREB signaling pathways， increasing the number and distribution of dendritic spines and repairing damaged neurons in the DP model rat's hippocampus.
Keywords：depression;Chaihu Yueju decoction;ethology;cAMP responsive element binding protein（CREB）;synapse associated protein
Abstract：ObjectiveThis study intends to study the regulatory effect and mechanism of the effective components of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos on inflammatory factors related to cerebral ischemia-reperfusion injury in rats through multiple levels of neuropathology， molecular neurobiology and functional behavior.MethodThe 32 male rats were randomly divided into four groups： sham group， model group， Danhong components compatibility group（720 mg·kg-1）， nimodipine （0.5 mg·kg-1）groups，each group of eight male rats.Cerebral ischemia was established by middle cerebral artery occlusion （MCAO） approach. The treatment was performed immediately and at 6 hour after MCAO.Hematoxylin-eosin （HE）staining was used to check the changes of brain histopathology， immunohistochemistry and Real time polymerase chain reaction （Real-time PCR） were used to check the expression of IL-1β and Nrf2 in brain tissue，Western blot was used to detect the protein expression of Nrf2 in brain tissue. The aim is to investigate the treatment mechanism of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos components in a rat model of cerebral ischemic-reperfusion injury.ResultHE staining results showed， compared with sham group， the surviving neurons amount in the model group was significantly reduced（P<0.01），compared with the MCAO group，the number of surviving neurons in the brain tissue of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos component compatibility group and nimodipine group was significantly increased（P<0.05，P<0.01）.The results of immunohistochemistry and Real-time PCR showed that，compared with normal group，IL-1β and Nrf2 expression in model group were significantly increased （P<0.01），compared with MCAO group， the expression of IL-1β and Nrf2 in Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos component compatibility group and the nimodipine group was significantly decreased （P<0.05，P<0.01）. Western blot results showed that， compared with sham group， Nrf2 positive expression in model group was much more increased （P<0.01）， compared with MCAO group， the expression of Nrf2 in Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos component compatibility group and the nimodipine group was significantly decreased （P<0.01）.ConclusionThe combination of effective components of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos can significantly down-regulate the expression of IL-1β and Nrf2 proteins.The mechanism is to activate the protein expression of inflammatory pathways， reduce the apoptosis of nerve cells， and finally inhibit the inflammatory response in the process of ischemic stroke injury.
Keywords：Salviae Miltiorrhizae Radix et Rhizoma;Carthami Flos;component compatibility;cerebral ischemic stroke;inflammatory reaction
Abstract：ObjectiveQuantitative analysis of anti-inflammatory synergistic pharmacodynamics mechanism of baicalin and wogonoside by medium efficiency principle.Methodinflammatory cell model was constructed by stimulating RAW264.7 cells by lipopolysaccharide （LPS） 100 μg·L-1 in vitro. The experiment was performed in the normal group， the model group， the andrographolide group （10 μmol·L-1）， the baicalin group （2.06，4.13，8.25，16.5，33，66，132 μmol·L-1） and the wogonoside group （2.94，5.88，11.75，23.5，47，94，188 μmol·L-1） and the baicalin-wogonoside combination group ［（2.06+2.94）（4.13+5.88）（8.25+11.75）（16.5+23.5）（33+47）（66+94）（132+188） μmol·L-1］. The levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in the cell culture supernatants after drug intervention for 50 min and 4 h were detected by enzyme-linked immunosorbent assay （ELISA） method. The level of nitric oxide （NO） in the cell culture supernatant after drug intervention for 24 h were detected by Griess method. Western blot was used to detect the activation levels of phosphorylation of nuclear factor-κB p65（p-NF-κB p65） and inducible nitric oxide synthase（iNOS） in cells after drug intervention for 2 h and 12 h. The fa/fu-dose profile of each indicator was drawn to observe the increase or decrease of effect.ResultCompared with normal group， the expression of p-NF-кB p65， iNOS and cytokines including TNF-α， IL-6 and NO （P<0.05，P<0.01） in the model group were significantly up-regulated. Compared with the model group， each group at high doses could inhibit the phosphorylation of NF-кB p65 protein（P<0.05），the baicalin group and the combined group could down-regulate the expression of iNOS protein in a concentration-dependent manner（P<0.01） and the baicalin group had no obvious inhibitory effect. each administration group at high dose could significantly inhibit the production of NO（P<0.05），but each group had no inhibitory effect on IL-6 production. The baicalin group and the combined group could significantly Inhibit the production of TNF-α（P<0.05） and there was no significant difference between the baicalin group and the model group. At the experimental dose， the fa/fu-dose table showed that the fa/fu value of p-NF-кB p65 and IL-6 in the combined group was not greater than the baicalin group and the wogonoside group. The fa/fu value of iNOS， TNF-α and NO in the combined group is higher than the baicalin group and the wogonoside group.ConclusionThe baicalin and wogonoside have different effects on different targets in the NF-κB pathway. The wogonoside is the main pharmacological substance in this combination and the combination shows different degrees of synergy or antagonism effects on different targets.
Abstract：ObjectiveTo investigate the effects of naringenin on oxidative stress and Tau protein phosphorylation of adrenal pheochromocytoma（PC12） cells injured by β-amyloid（Aβ）25-35 and its relationship with estrogen receptor（ER） and phosphatidylinositol -3 kinase/protein kinase B（PI3K/Akt） signaling pathway.MethodThe PC12 cells were intervened with Aβ25-35 to prepare the injury model. The experiment was divided into blank group， model group， naringenin（400，40，4，0.4，0.04，4×10-3，4×10-4，4×10-5 μmol·L-1）group， positive drugs estradiol（E2）（1 nmol·L-1）+Aβ25-35 group， naringenin（0.4，0.04，4×10-3，4×10-4，4×10-5 μmol·L-1）+Aβ25-35 group， E2+Aβ25-35+ER antagonist（ICI182780）（1 μmol·L-1） group， naringenin+Aβ25-35+ICI182780 group， E2+Aβ25-35+PI3K blocker（LY294002）（50 μmol·L-1） group， naringenin+Aβ25-35+LY294002 group. Methye thiazolye telrazlium（MTT）method was used to detect the cell proliferation index， 2'，7'-Dichlorodi -hydrofluorescein diacetate（DCFH-DA） was used as a fluorescent probe to detect the content of reactive osygen species（ROS）， the content of malondialdehyde（MDA） and the activity of superoxide dismutase（SOD） were measured by thiobarbituric acid（TBA） and oxidase methods， Western blot was used to detect the expression of phosphorylated Tau protein/total Tau protein（p-Tau/t-Tau）.ResultAccording to the results of MTT experiment， 0.4μmol·L-1 was selected as the best effective concentration of naringenin， compared with the blank group， the cell proliferation index of model group decreased significantly （P<0.01）， compared with model group， the cell proliferation index of naringenin+Aβ25-35 group increased significantly （P<0.01）. In addition， compared with blank group， the content of ROS， MDA and the expression of p-Tau/t-Tau in the model group increased significantly （P<0.01）， and the activity of SOD decreased significantly （P<0.01）， compared with model group， the content of ROS， MDA and the expression of p-Tau/t-Tau in naringenin+Aβ25-35 group decreased significantly （P<0.01）， and the activity of SOD increased significantly （P<0.01）， compared with naringenin+Aβ25-35 group， the addition of ICI182780 and LY294002 significantly reversed the role of naringenin in the above indicators （P<0.01）. The effect of naringenin was similar to that of E2.ConclusionNaringenin can improve the cell proliferation index and protect PC12 cells from Aβ25-35 injury， which may be achieved by activating ER and PI3K/Akt signaling pathway to reduce ROS， MDA content， p-Tau/t-Tau expression and promote SOD activity.
Abstract：ObjectiveTo investigate the anti-tumor effect mechanism of atractylenolide Ⅱ by studying its effect on macrophage polarization.MethodPhorbol myristate acetate （PMA） was used to induce THP-1 cells differentiation into macrophages， and methylthiazolyldiphenyl-tetrazolium bromide（MTT）colorimetric assay was used to detect the effect of different concentrations of atractylenolide Ⅱ on macrophage growth at different time points to screen out the safe concentration of atractylenolide Ⅱ. The macrophages were treated with different concentrations of atractylenolide Ⅱ for 24 hours and then were co-cultured with gastric cancer cells. The survival of the two types of cells was observed under light microscope. The proliferation of gastric cancer cells was detected by MTT assay to determine the effective administration concentrations of atractylenolide Ⅱ. Cells were divided into blank group， model group， atractylenolide Ⅱ high dose group （200 mg·L-1）， atractylenolide Ⅱ medium dose group （100 mg·L-1）， and atractylenolide Ⅱ low dose group（50 mg·L-1）. Wound healing assay was carried out to observe the effects of different concentrations of atractylenolide Ⅱ on the migration and morphology of gastric cancer cells. The expression levels of M1 and M2 macrophage surface markers CD86 and CD206 were detected by flow cytometry analysis（FCM）. Quantitative polymerase chain reaction（Real-time PCR）and Western blot were used to detect M1， M2 macrophage-associated tumor necrosis factor （TNF） -α， human leukocyte antigen 2 （HLA-DRA）， CD80， transforming growth factor （TGF）-β， interleukin （IL） -10 and IL-6 genes and protein expression. Western blot was used to detect intracellular phosphatidyl inositol kinase （PI3K） and p-PI3K protein expression in macrophages.ResultWhen the concentration of atractylenolide Ⅱ was 1， 10， 50， 100， 200 mg·L-1， it showed no inhibition on macrophage growth. As compared with the model group， macrophages treated with 50， 100， 200 mg·L-1 atractylenolide Ⅱ significantly inhibited tumor cell proliferation （P<0.01）. As compared with the model group， the migration rate of tumor cells in the atractylenolide Ⅱ （200，100 mg·L-1） groups decreased （P<0.05）. The expression levels of CD86 on M1 macrophage surfacen in the atractylenolide Ⅱ （200，100，50 mg·L-1） groups were increased（P<0.05，P<0.01）， and the expression levels of CD206 on M2 macrophagen in the atractylenolide Ⅱ （200 mg·L-1） group were decreased （P<0.05）. The expression levels of M1 macrophage-associated cytokines TNF-α， HLA-DRA， CD80 mRNA in the atractylenolide Ⅱ （200，100 mg·L-1） groups were increased（P<0.05，P<0.01）， and TNF-α protein expression in the atractylenolide Ⅱ （200 mg·L-1） group was increased （P<0.05）， M2 type macrophage-associated cytokine TGF-β mRNA expression levels in the atractylenolide Ⅱ （50 mg·L-1） group were decreased， and IL-10， IL-6 protein expression levels in the atractylenolide Ⅱ （200 mg·L-1） group were decreased （P<0.05，P<0.01）. The expression levels of p-PI3K protein in the atractylenolide Ⅱ （200，100 mg·L-1） groups were also decreased（P<0.05，P<0.01）.ConclusionAtractylenolide Ⅱ could induce the polarization of macrophages to M1 type by reducing the expression of p-PI3K in macrophages and inhibiting the proliferation and migration of gastric cancer cells.
Abstract：ObjectiveTo explore the correlation between the occurrence of colorectal cancer and different internal environment （cold and heat）.MethodThe 70 Wistar rats （male and female） were randomly divided into blank group （10 cases）， cold model group （30 cases） and heat model group （30 cases）. The cold syndrome model was made by intragastric infusion of cold water （0 ℃） and soaking in cold water （10 ℃）. The heat model was made by ethanol （30%） and capsaicin solution （0.9 g·L-1）. The blank group was given normal saline by gavage， 10 mL·kg-1·d-1， for 5 consecutive weeks. The colorectal cancer model was made by subcutaneous injection of DMH solution in the back of neck in the cold model group and heat model group at the 6th week， 25 mg·kg-1，once a week，for 12 consecutive times. During the carcinogenesis， only 30% ethanol solution was given to the heat model group， and the modeling was maintained in cold model group， 10 mL·kg-1·d-1， for 38 weeks. The general state of the rats in each group was observed， and the changes of food intake and body weight were measured. At the 27th， 29th， 32th， 35th and 38th weeks， samples were collected in batches. Intestinal tissues were subjected to hematoxylin-eosin staining （HE） and detection of sodium， potassium-adenosine triphosphate （Na+ K+-ATP）， calcium， magnesium-adenosine triphosphate （Ca2+ Mg2+-ATP） activity， lactate dehydrogenase （LDH） activity and succinate dehydrogenase （SDH） activity.ResultThe symptoms of hematochezia and ascites in cold model group were earlier than those in heat model group. As compared with the blank group， the food intake and body weight were decreased in cold model group and heat model group. As compared with the blank group， the length of the large intestine was shorter in cold model group at the 32nd and 35th week （P<0.05）， the activities of Na+ K+-ATP， Ca2+ Mg2+-ATP increased significantly in heat model group at the 27th， 29th and 38th week （P<0.05， P<0.01）， the SDH enzyme activity was decreased in the cold model group at the 29th and 35th week （P<0.05， P<0.01）， the SDH enzyme activity was significantly increased in the heat model group at the 38th week （P<0.01）. At the 27th week， LDH enzyme activity was significantly reduced in cold model group （P<0.01）. The LDH activity was increased significantly in heat model group at 29th and 32nd week （P<0.05， P<0.01）. As compared with the heat model group， the large intestine texture of the cold model rats showed greater brittleness， aggravated fibrosis， more obvious fibroproliferative characteristics， higher tumor incidence， and more serious tumor differentiation. The Na+ K+-ATP， Ca2+ Mg2+-ATP enzyme activities of the cold model rats were significantly reduced at 27th， 29th， and 38th weeks （P<0.01）， the SDH enzyme activity was significantly reduced in the cold model rats at 29th and 38th weeks （P<0.01）， the LDH activity was reduced in the cold model group at 32nd week （P<0.05）.ConclusionCold environment for colorectal cancer promotes tumorigenesis， and the hot environment can also promote tumorigenesis in a later stage.
Keywords：cold and heat syndrome;colorectal cancer model;adenosine triphosphate （ATPase）;lactate dehydrogenase （LDH）;succinate dehydrogenase （SDH）
Abstract：ObjectiveBy the method of network pharmacology， the mechanism of Pinelliae Rhizoma and Citri Exocarpium Rubrum in the treatment of metabolic syndrome was explored.MethodEffective components of Pinelliae Rhizoma and Citri Exocarpium Rubrum were retrieved by TCMSP database，and then selected by ADME parameters. TCMID，BATMAN-TCM，SymMap，TCM-MESH database were used to supplement effective components of TCMSP. TCMSP target prediction model was used to predict potential targets of drugs. DRUGBANK，DisGeNET，CTD，GeneCards，OMIM，PharmGkb，KEGG，DiGSeE databases were retrieved to obtain the targets of metabolic syndrome，and the chips were downloaded and analyzed through GEO database No.GSE98895 to screen out the differential genes of normal people and patients with metabolic syndrome，and supplement the target databases of metabolic syndrome. The intersections of Pinelliae Rhizoma-Citri Exocarpium Rubrum and metabolic syndrome disease targets were obtained by Rstudio 3.6.2. The above intersection targets were imported into the Metascape database for module analysis and overall GO（Biological Process），KEGG and Reactome pathway analysis. The core targets were selected from the intersection targets by using the cytohubba plug-in，the core genes were input into the database of BioGPS，Genecards to analyze the tissue distribution and subcellular distribution，and the core targets were assigned by using the database of DisGeNET.ResultA total of 34 active components and 120 targets of Pinelliae Rhizoma and Citri Exocarpium Rubrum were screened out，and 115 targets were obtained after the intersection of Rstudio 3.6.2. The results of Metascape module analysis and whole analysis were mostly the same，mainly involving the biological processes， such as ligand receptor interaction，calcium ion，cGMP-PKG，cholinergic synapse，thyroid hormone，insulin. The cytohubba plug-in was used to screen out 17 targets，involving 17 key genes， such as VEGFA and NOS3. The tissue and subcellular distribution of the core targets mainly included lymphoblasts，CD33+_myeloid cells，amygdala，pineal and cytoplasmic matrix，mitochondria. The main proteins were signal molecules，kinases and nucleic acids.ConclusionPinelliae Rhizoma and Citri Exocarpium Rubrum could treat metabolic syndrome through complex biological processes and pathways，such as blood circulation，ligand receptor interaction of nerve activity，cGMP-PKG，interleukin-related action，calcium ion. This indicates that traditional Chinese medicine（TCM） treated diseases through multi-component，multi-target，multi biological processes，multi-channel and other ways（which is also proved by the distribution of core genes in the tissue，subcellular and protein ascription information），indicating the superiority of the holism concept of TCM. Erchetang and its similar prescriptions are suitable for treating metabolic syndrome，which also indicates that the principle of "treating different diseases with the same therapy" of TCM is not only reflected at the theoretical level； and network pharmacology needs to be further proved in the combination with experimental verification.
Abstract：ObjectiveTo observe the efficacy of modified Sanxue Gegentang combined with nerve block in treating postherpetic neuralgia due to blood stasis syndrome， and explore its possible mechanism.MethodThe 120 patients were randomly divided into control group and observation group， with 60 cases in each group. The patients in control group received Dahuang Zhechong tablet + nerve block， and the patients in observation group received modified Sanxue Gegentang + nerve block for 4 weeks. The clinical symptoms （simplified McGill pain scale， overall impression change scale， traditional Chinese medicine symptoms）， serum medium pain ［endothelin-1 （ET-1）， prostaglandin E2 （PGE2）， substance P （SP）， cydooxygenase-2 （COX-2）］， blood viscosity index ［high blood viscosity （HBV）， medium blood viscosity （MBV）， low blood viscosity （LBV）， and plasma viscosity （PV）］ were observed before and after treatment. The clinical efficacy， recurrence rate in 12 months follow-up， and the incidence of adverse reactions were compared between two groups.ResultThe 5 cases in control group and 1 case in the observation group were lost during the study. The total effective rate in observation group was 96.6%（57/59）， higher than 74.5% （41/55） in control group （χ2=5.729， P<0.05）. As compared with control group after treatment， the McGill pain scale， overall impression change rating scale， main TCM symptom scores， pain medium （ET-1， PGE2， SP， COX-2）， and blood viscosity （HBV， MBV， LBV， PV） were significantly lower in observation group （P<0.05）. In the follow-up for 12 months， the recurrence rate was 7.0% （4/57） in observation group， lower than 43.9% （18/41） in control group （χ2=7.294， P<0.05）. During the study period， the incidence of adverse reactions was 8.5% （5/59） in observation group and 7.3% （4/55） in control group， without significant difference between two groups.ConclusionModified Sanxue Gegentang combined with nerve block can significantly improve the clinical symptoms of patients with postherpetic neuralgia due to blood stasis syndrome， showing a low recurrence rate incidence of adverse reactions.
Abstract：ObjectiveTo observe the clinical effect of Kangfuxin solution combined with Almagate suspension on the complication of gastroscope biopsy.MethodTotally 276 cases of chronic superficial gastritis gastroscope biopsy were divided into treatment group and control group randomly. Treatment group was treated by Kangfuxin solution combined with Almagate suspension，while the control group was given no precautionary measures as usual. Then the patients received the abdominal symptom score and fecal occult blood test（FOBT）after 1 week.ResultThe cases with abdominal pain，burning sensation and stool occult blood in the control group were more than the treatment group significantly（P<0.05）. The patients' abdominal symptom score in control group was higher than that before the gastroscope biopsy，and that was lower in treatment group（P<0.05）. And the patients' abdominal symptom score of the control group was significantly higher than that of the treatment group after the gastroscope biopsy（P<0.05）. Compared with the Helicobacter pylori（HP）-infected subgroup，the incidences of early satiety and ventosity in the control group were higher than those of the treatment group（P<0.05）. The abdominal symptom score of the patients infected with helicobacter pylori（HP）in control group was significantly higher than that before the gastroscope biopsy and in the treatment group（P<0.05）.ConclusionKangfuxin solution combined with Almagate suspension is safe and effective in preventing complications of gastroscope biopsy，especially for patients infected with HP. The method could avoid exacerbating clinical symptoms.
Abstract：ObjectiveTo observe the clinical efficacy of compound Huanggen granules combined with Entecavir tablets for patients with chronic hepatitis B with syndrome of collateral retardation due to blood stasis.MethodThe 130 patients with chronic hepatitis B with syndrome of collateral retardation due to blood stasis were randomly divided into observation group and control group by using random number table method. The 65 patients in observation group were treated with compound Huanggen granules combined with Entecavir tablets， and 65 patients in control group were treated with Entecavir tablets orally. The changes of liver function ［alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin（TBIL）， albumin（Alb）］ ， coagulation function ［prothrombin time（PT）， activated partial thromboplastin time（APTT）， thrombin time（TT）， fibrinogen（FIB）］， serum Hepatitis B Virus DNA（HBV DNA） levels， hepatitis B e antigen quantification（HBeAg）， stiffness of the liver， liver imaging （portal vein width， spleen thickness）， liver histopathology knodell HAI classification for chronic hepatitis， and changes in ishak fibrosis score at 24，48 weeks were observed in both groups.ResultAfter 48 weeks， the indexes of the two groups were improved to different degrees （P<0.05，P<0.01）， serum HBV DNA negative conversion rate， liver function， coagulation index， liver stiffness， portal vein diameter， Knodell HAI grade portal inflammation， interface activity， hepatic lobular activity， and Ishak fibrosis score in observation group were better than those in the control group （P<0.05）. Patients in the control group had no significant improvement in liver lobular activity after treatment in the Knodell HAI classification of chronic hepatitis.ConclusionCompound Huanggen granules combined with Entecavir tablets are more effective in treating patients with chronic hepatitis B with syndrome of collateral retardation due to blood stasis than Entecavir tablets alone. The combination of two drugs helps to improve the rapid response rate of HBV DNA， and has a better effect on improving liver function， controlling the development of liver fibrosis and preventing related complications.
Keywords：compound Huanggen granules;Entecavir tablets;chronic hepatitis B;syndrome of collateral retardation due to blood stasis;clinical observation
Abstract：ObjectiveTo observe the clinical efficacy of addition and subtraction therapy of Shashen Maidongtang for syndrome of Yin deficiency and lung heat at convalescent period in patients with mycoplasma pneumoniae pneumonia， and to investigate its regulatory effect on immune inflammatory response.MethodOne hundred and forty-eight patients were randomly divided into control group （74 cases） and observation group （74 cases） by random number table. During the treatment， 70 patients completed the study in control group （loss to follow-up in 2 cases， elimination in 3 cases）， and 71 patients completed the study in observation group （loss to follow-up in 4 cases）. Patients in both groups got Azithromycin tablets for 3 days， 10 mg·kg-1·d-1， pidotimod granules， 0.4-0.8 g/time， 2 times/day， and Ambroxol oral solution， 2.5-15 mL/time， 2 times/day. The patients in control group additionally received Yangyin Qingfei oral liquid， 5-10 mL/time， 2-3 times/day， while the patients in observation group additionally received addition and subtraction therapy of Shashen Maidongtang， 1 dose/day， with a treatment course of 7 days in both groups. Before and after treatment， scores of cough symptom and cough visual analogue score （VAS） were graded， and the time to disappearance of cough， lung rales and expectoration as well as the time to recovery of chest radiograph were recorded. Scores of syndrome of yin deficiency and lung heat and Leicester Cough Questionnaire （LCQ） were also graded. Levels of T lymphocyte subsets （CD3+， CD4+， CD8+ and CD4+/CD8+）， interleukin-6 （IL-6）， IL-10， tumor necrosis factor-α （TNF-α）， substance P （Substance P） and procalcitonin gene related peptide （CGRP） were detected， and the safety was also evaluated.ResultTotal effective rate for the disease was 95.77% （68/71） in observation group， higher than 82.86% （58/70） in control group （χ2=6.186， P<0.05）. Scores of cough symptoms and VAS were higher than those in control group （P<0.01）， and scores of various items in LCQ scale and the total score of LCQ were all higher than those in control group （P<0.01）. The time to disappearance of cough， lung rales and expectoration as well as the time to recovery of chest radiograph in observation group were shorter than those in control group （P<0.01）. Levels of CD3+， CD4+ and CD4+/CD8+ were higher than those in control group （P<0.01）， and levels of CD8+， IL-6， IL-10， TNF-α， SP in serum and CGRP in sputum were all lower than those in control group （P<0.01）.ConclusionOn the basis of conventional western medicine treatment， addition and subtraction therapy of Shashen Maidongtang can reduce the degree of cough， shorten the course of disease， improve the quality of life， improve the cellular immune function， reduce the inflammatory injury， and reduce the sensitivity of cough， with better clinical efficacy and safety as compared with Western medicine alone.
Keywords：mycoplasma pneumoniae pneumonia;convalescent period;syndrome of Yin deficiency and lung heat;Shashen Maidongtang;immune function;inflammatory factor;procalcitonin gene related peptide;substance P
Abstract：ObjectiveTo observe the clinical efficacy of addition and subtraction syndrome differentiation method of Huanglian Ejiaotang and Baihe Dihuangtang in the treatment of menopausal syndrome （MPS） and symptoms of deficiency of yin and hyperactivity of fire， and investigate its regulation effects on neuroendocrine network.MethodOne hundred and forty-eight patients were divided into control group （74 cases） and observation group （74 cases） according to the random number table method. The patients in control group received estradiol/norethisterone acetate tablets， 3 mg/time， 1 time/day， while the patients in observation group received addition and subtraction of Huanglian Ejiaotang and Baihe Dihuangtang based on the same western medicine treatment as control group， orally 1 dose/day， with a treatment course of 3 months in both groups. Kupperman index （KI） score was evaluated before treatment and at 1 month， 2 months and 3 months after treatment. The Menopausal Quality of Life （MENQOL）， self-rating depression scale （SDS）， Self-rating Anxiety Scale （SAS）， Pittsburgh Sleep Quality Index （PSQI） and score of deficiency of yin and hyperactivity of fire were evaluated before and after treatment. The levels of serum estradiol （E2）， follicle stimulating hormone （FSH）， luteinizing hormone （LH）， norepinephrine （NE）， 5-hydroxytryptamine （5-HT）， nitric oxide （NO）， endothelin-1 （ET-1） and calcitonin gene-related peptide （CGRP） were detected before and after treatment.ResultThe clinical efficacy in observation group was better than that in control group （Z=2.115， P<0.05）， and the KI scores in observation group at 1 month， 2 months and 3 months after treatment were lower than those in control group （P<0.01）， and the decrease of KI score in observation group after treatment was larger than that in control group （P<0.01）. The scores of various dimensions of MENQOL scale in observation group were lower than those in control group （P<0.01）. The scores of deficiency of yin and hyperactivity of fire， SAS， SDS and PSQI in observation group were all lower than those in control group （P<0.01）， and the level of E2 in observation group after treatment was higher than that in control group （P<0.01） ，while the levels of FSH and LH were lower than those in control group （P<0.01）.The levels of 5-HT and NE in observation group were higher than those in control group （P<0.01）.The NO level in observation group was higher than that in control group （P<0.01）， while the levels of CGRP and ET-1 were lower than those in control group （P<0.01）. During the study period， there were 5 drop-out cases， 2 excluded cases， and 67 completed cases in the control group. There were 7 drop-out cases， 1 excluded case， 66 completed cases in the observation group.ConclusionOn the basis of hormone replacement therapy， addition and subtraction of Huanglian Ejiaotang and Baihe Dihuangtang for patients with MPS （with symptom of deficiency of yin and hyperactivity of fire） can improve clinical symptoms， relieve depression and anxiety， improve sleep quality and enhance quality of life， and its clinical efficacy is superior to hormone replacement therapy alone. It can also regulate endocrine hormones， vasomotor factors and monoamine neurotransmitters， thereby improving clinical symptoms.
Keywords：menopausal syndrome;symptom of deficiency of Yin and hyperactivity of fire;Huanglian Ejiaotang;Baihe Dihuangtang;endocrine hormones;vasomotor factors;monoamine neurotransmitters
Abstract：ObjectiveTo study the serum pharmacochemistry of Aurantii Fructus （AF）， and to investigate the pharmacological material basis of AF extract in rats.MethodRapid identification and speculation of the prototype constituents and their metabolites in vivo were carried out according to the relative retention time， accurate relative molecular mass， cleavage fragments of MS/MS and neutral loss of metabolites with ultrahigh performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry （UPLC-Q-TOF/MS） technique by comparing the differences between different samples such as AF extracts， blank plasma， and administered plasma under the same chromatographic and mass spectrometric conditions.ResultAfter oral administration of the AF extract， 74 transitional constituents absorbed into the blood were detected in serum， in which 49 compounds were prototype constituents and the other 25 were metabolites. The prototype constituents could be divided into dihydroflavones， polymethoxyflavonoids， limonins， coumarins and alkaloids. The identified metabolites included glucuronic acid conjugates， sulfuric acid conjugates， hydroxylated products of flavonoid glycosides and polymethoxyflavonoids， as well as the simultaneous glucuronidation and sulfation products.ConclusionThe constituents absorbed into the blood and their metabolites may be the pharmacodynamic components of AF. Among them， alkaloids， polymethoxyflavonoids and coumarins are mainly introduced into the blood in the prototype form， while naringin and neohesperidin （the index components） exert effect mainly through hydrolysis into aglycones. This work will help to further elucidate the material basis of AF.
Keywords：Aurantii Fructus;serum pharmacochemistry;ultrahigh performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry （UPLC-Q-TOF/MS）;prototype components;metabolites;dihydroflavones;polymethoxyflavonoids
Abstract：ObjectiveA systematical study on the anti-breast cancer mechanism of tryptanthrin in breast cancer-bearing mice was done by Label-free proteomics.MethodUPLC-MS was used to detect the expressed-proteins of tryptanthrin inhibiting breast cancer in mice， chromatographic separation was achieved on the Ionoptics nano UPLC C18 column （0.075 mm×250 mm， 1.6 μm）， and gradient elution was performed with 0.1% formic acid aqueous solution-0.1% formic acid acetonitrile solution as mobile phase. Data acquisition was carried out in electrospray ionization （ESI） under the positive ion mode， the scanning range was m/z 100-1 700， MaxQuant 220.127.116.11 was used for database retrieval. Label-free proteomics with high resolution mass spectrometry was used to screen differentially expressed proteins between the model group of 4T1 breast cancer mice and oral administration group of tryptanthrin （100 mg·kg-1）. The proteomics of tryptanthrin against breast cancer was carried out.ResultA total of 3 997 proteins were identified in this proteomics research， and 2 911 proteins were quantifiable. A total of 750 differentially expressed proteins were identified between the model group and the tryptanthrin group， 286 proteins were up-regulated and 464 proteins were down-regulated. Gene ontology analysis showed that these differentially expressed proteins were mainly involved in biological processes of proliferation， cell migration， apoptosis， immunity， angiogenesis， inflammatory regulation， etc. Kyoto encyclopedia of genes and genomes pathway analysis further indicated that these proteins were mainly concentrated in T cell receptors， B cell receptors， Toll-like receptors， nuclear transcription factor-κB （NF-κB）， Ras proteins， interleukin-17， tumor necrosis factor， phosphatidylinositol 3-kinase/protein kinase B （PI3K-Akt）， mitogen-activated protein kinase （MAPK） and other signaling pathways.ConclusionThe differentially expressed proteins closely related to anti-breast cancer effect of tryptanthrin on 4T1 breast cancer mice are effectively screened out， including up-regulating proteins of leukocyte differentiation antigen 14 （CD14）， prostaglandin G/H synthase 2 （PTGS2）， E3 ubiquitin-protein ligase and down-regulating proteins of CD44， heat shock 70 kDa protein 1A （HSPA1A）， macrophage migration inhibitory factor （MIF）， NF-κB， ribosomal protein S6 kinase alpha-4 （RPS6KA4） and high mobility group protein B1 （HMGB1）. These findings suggest that tryptanthrin can inhibit breast cancer in mice mainly through regulating tumor inflammatory microenvironment.
Abstract：ObjectiveTo investigate the moisture adsorption and thermodynamic characteristics of raw products， wine-processed products and fried charcoal products of Rhei Radix et Rhizoma， in order to guide their drying and storage.MethodStatic isotherm weighing method was used to determine the adsorption isotherm curves of three Rhei Radix et Rhizoma decoction pieces at 25， 35， 45 ℃， and the test data were fitted with 7 commonly used water adsorption models to determine the best model for studying the adsorption thermodynamic parameters of these decoction pieces.ResultThe best adsorption models of these three decoction pieces were all GAB model. At 25， 35， 45 ℃， the absolute safe moisture content of fried charcoal products was 7.43%， 6.79% and 6.20%， of wine-processed products was 8.68%， 8.17% and 7.03%， of raw products was 9.88%， 9.36% and 7.77%， respectively. At 25， 35， 45 ℃， the relative safe moisture content of fried charcoal products was 9.46%， 8.63% and 8.21%， of wine-processed products was 11.49%， 11.03% and 9.74%， of raw products was 13.49%， 12.66% and 11.14%， respectively. The net equivalent heat of adsorption （Qst） and differential entropy （Sd） of these three kinds of decoction pieces all decreased with the increase of equilibrium moisture content， Qst and Sd were in accordance with the entropy-enthalpy complementary theory. The constant velocity temperatures of raw products， wine-processed products and fried charcoal products of Rhei Radix et Rhizoma were 386.66， 391.15， 394.34 K （unit conversion of 1 K=-272.15 ℃）， their Gibbs free energies were 0.372 2， 0.406 0， 0.372 2 kJ·mol-1， respectively. Their adsorption processes were an unspontaneous process driven by enthalpy.ConclusionThe orders of equilibrium moisture content， monomolecular layer moisture content， Qst and Sd of three Rhei Radix et Rhizoma decoction pieces are all raw products>wine-processed products>fried charcoal products. The moisture absorption capacity of the decoction pieces is ranked as raw products>wine-processed products>fried charcoal products. The frying and roasting process significantly affects the hygroscopicity and thermodynamic properties of the three decoction pieces， the reason for this difference may be that the high temperature of the stir-frying results in the decrease of the hygroscopic groups and the increase of the hydrophobic materials in raw products， and the change in the texture of the decoction pieces. The research on the water adsorption characteristics of three Rhei Radix et Rhizoma decoction pieces can provide reference for selecting their storage conditions and drying process.
Keywords：decoction pieces;raw products of Rhei Radix et Rhizoma;wine-processed products of Rhei Radix et Rhizoma;fried charcoal products of Rhei Radix et Rhizoma;adsorption isotherm curve;thermodynamic properties;net equivalent heat of adsorption
Abstract：ObjectiveTo investigate the chemical constituents from the ethyl acetate extract of 95% ethanol extract from Ajuga ovalifolia var. calantha.MethodA. ovalifolia var. calantha was percolated with 95% ethanol，and the percolate was concentrated under reduced pressure to obtain the extract. The extract was then dispersed with water and extracted with petroleum ether to obtain the petroleum ether extract fraction and water layer. Then ethyl acetate was used to extract the water layer to obtain the ethyl acetate extract fraction. The Compounds from the ethyl acetate extract fraction were isolated and purified by silica gel，Sephadex LH-20，MCI column，ODS column chromatography and semi-preparative high performance liquid chromatography. Their structures were elucidated by interpretation of NMR，ESI-MS and other spectral evidence.Result17 Compounds were isolated and elucidated as bakkenolide-E（1），loliotide（2），isololiotide（3），（E）-linalool-1-oic acid（4），umbelliferone（5），phillygenin（6），1-（4-hydroxyphenyl）-ethanol（7），（2E）-8-hydroxy-2，6-dimethyl-2-octenoic acid（8），1H-indole-3-carboxylic acid（9），ajugarin I（10），ajugalactone（11），luteolin（12），20-hydroxyecdysone-2-acetate（13），benzyl-4'-hydroxy-benzoyl-3'-O-β-D-glucopyranoside（14），harpagide（15），6-deoxy-8-acetylharpagide（16），and acteoside（17）.ConclusionCompounds 1，3-4，6-9，14 were isolated from the plants of Ajuga for the first time，and compounds 2，5，10，13，15-16 were isolated from this plant for the first time.
Keywords：Ajuga ovalifolia var. calantha;ethyl acetate extract;chemical constitutes;structure identification
Abstract：ObjectiveTo propose a new method for detecting and evaluating traditional Chinese medicine （TCM） by artificial intelligence and machine vision technology.MethodTaking Fritillariae Cirrhosae Bulbus， Crataegi Fructus and Pinelliae Rhizoma as the research objects， big data of pictures was collected by machine vision and the image database was established. Through the intelligent analysis of the external characteristics of TCM， the deep convolutional neural network model was established to realize the functions of location detection and variety identification by means of deep learning， so as to significantly improve the accuracy of rapid identification of TCM.ResultThe classification accuracy of 11 kinds of Chinese herbal pieces （raw， fried， parched and charred products of Crataegi Fructus， Pinelliae Rhizoma， Pinelliae Rhizoma Praeparatum Cum Zingibere et Alumine， Pinelliae Rhizoma Praeparatum， Pinelliae Rhizoma Praeparatum Cum Alumine and three products of Fritillariae Cirrhosae Bulbus） could be more than 99%， and the average recognition accuracy of specific categories could reach more than 97%.ConclusionThe intelligent identification technology of TCM decoction pieces realized by deep learning algorithms has the advantages of simplicity， rapidity， high precision and quantifiable detection， which can provide technical support for the quality detection and evaluation of TCM， and enrich the research ideas of quality evaluation of TCM.
Abstract：ObjectiveTo investigate the ecological environment and associated plants of mountain cultivated Panax ginseng were investigated in Xifeng county，Kuandian Manchu autonomous county and Huanren Manchu autonomous county of Liaoning province，in order to provided suggestions on the development of the mountain cultivated P. ginseng in these areas and necessary reference materials for the analysis of ecological suitability of mountain cultivated P. ginseng.MethodThe field survey was conducted through sample plot survey in the survey pattern ofbased on representative area-sample plot-quadrat，literature review and data collation， were conducted to record indicators， calculate relevant parameters and make summary.ResultThe associated plants were investigated based on 42 samples of mountain cultivated ginseng collected. A total of 95 plant species were identified. The correlation analysis was made on the six main species with an occurrence frequency of higher than 33%， and they were all positively correlated. Asarum heterotropoides var. mandshuricum was the strongest，which was followed by Schisandra chinensis， and Tilia tuan was the strongest in the arbor layer. The survey also found many problems in these areas，such as fewer large-scale planting bases， more scattered households，mixed germplasms，irregular planting techniques，simple planting mode and less sources of economic income generation.ConclusionXifeng county，Kuandian county and Huanren county of Liaoning province boast a superior ecological environment and abundant resources of mountain cultivated P. ginseng. The survey gets the whole picture of the ecological environment and associated plants of mountain cultivated P. ginseng in representative areas of Liaoning province，so as to provide the basis for the further development of mountain cultivated P. ginseng cultivation industry and the promotion of mountain cultivated Panax ginseng economy.
Abstract：Alpinia katsumadai is recorded in Chinese herbal classics of previous dynasties，with a long history of medicinal use and significantly efficacy. This paper made a comprehensive textual research and summarization of the name，origin，distribution of producing areas，genuine producing area，harvesting time，processing method，property and flavor，and treatment functions of A. katsumadai by reviewing the ancient and modern literatures systematically. A. katsumadai has many alias names， such as Doukou，Loukou，Caokou in Chinese. Through the analysis of ancient herbal researches and drawings，it is concluded that there was some disordered uses of A. katsumadai with A. zerumbet and Amomum tsaoko. And the varieties of A. katsumadai have changed in some areas from ancient to present. The original plants of A. katsumadai is Alpinia katsumadai， which belongs to Alpinia of Zingiberaceae in modern textual research. A. katsumadai mainly grows in Lingnan and some other tropical areas； especially， those produced in Hainan have a better quality. The harvest time is usually in summer and autumn when fruits are all ripe. The processing methods of A. katsumadai are various， including heating and bending wrapped with flour， processing slowly with Euodiae Fructus，stir-frying in ancient times，while purifying processing （peeling） is generally used in modern times. A. katsumadai has the effect in invigorating spleen and warming stomach，lower Qi and relieving stagnation-syndrome，drying dampness and driving cold. In modern studies， efforts shall be made to strengthen basic research，establish quantitative standards for processing and digital standard for genuine medicinal materials of A. katsumadai， and deeply explore the compatibility regularity and application of A. katsumadai in ancient prescriptions， in order to ensure the quality and maximize its medicinal value in modern studies.
Abstract：A good neural microenvironment is an important basis for improving the damaged nerves and promoting axonal repair and regeneration. The destruction of neural microenvironment， closely related to the lack of neurotrophic factors， microcirculation disorders and immune abnormalities， is the key pathogenesis leading to diabetic peripheral neuropathy （DPN）. In traditional Chinese medicine， disharmony between Ying and Wei is considered as the key pathology in the development of DPN. It may be manifested as Ying and Wei deficiency， or Ying and Wei impassability， or Ying， Wei， Qi and blood intersection disorders， all of which may cause body fluid condensed into phlegm， blood into blood stasis， further leading to the mutual knot of phlegm and blood stasis， meridian obstruction， numbness and pain of limbs. "Regulating Ying and Wei and tonifying spleen and stomach" is the main therapeutic idea to promote intersection between Ying and Wei and unblock Qi and blood. The method has a significant effect on DPN. However， the current studies on the mechanism of regulating Ying and Wei in the treatment of DPN are still in lack of in-depth discussion， and the studies are mostly limited to the microcirculation disorders. Numerous studies have confirmed that the courses and distribution， physiological characteristics， functions of Ying and Wei are closely related to nerve， immune， metabolic substances and microcirculation. Based on the modern medicine essence of Ying and Wei， the author thinks that the discussion on connotation of the Ying and Wei from the perspective of neural microenvironment has a scientific basis， and regulating Ying and Wei is not only inherited from the traditional Chinese medicine theory， but also conforms to the modern understanding on DPN pathogenesis and treatment. Regulating Ying and Wei and smoothing middle-jiao can improve neural microenvironment and give play to the role of restoring damaged nerve， and its mechanism may be related to regulating neurotrophic factors， immune active substances， metabolites， and microcirculation dysfunction.
Keywords：regulating Ying and Wei;neural microenvironment;diabetic peripheral neuropathy;Ying and Wei theory
Abstract：Texture analyzer is a multifunctional physical property analyzer. Through a variety of test modes such as compression， puncture， shearing and stretching， hardness， adhesion， elasticity， cohesiveness and other physical property parameters are characterized. The results are objective， sensitive， and accurate， which can provide theoretical basis and reference for interpretation of basic theoretical， optimization of prescription technology and quality control research of pharmaceutical preparations. Firstly， the principles， test modes， measurement indexes and measurement data analysis methods of texture analyzer were summarized in this paper. Secondly， the application progress of texture analyzer in solid preparations， semi-solid preparations， liquid preparations， traditional Chinese medicine decoction pieces and their intermediates was described in detail， and compared with the commonly used measurement methods of physical property indicators. Finally， the application prospects of texture analyzer and its research contents to be improved were reviewed， so as to facilitate the use of texture analyzer by pharmaceutical researchers to promote the development of related fields of pharmaceutics， and to provide new ideas and methods for the research and development of pharmaceutical preparations.
Keywords：texture analyzer;test modes;pharmaceutical preparations;physical parameters;formulation screening;quality control;traditional Chinese medicine decoction pieces