Abstract：Objective:To explore the effect of Da Chengqitang on the lung index, lung index inhibition rate, lung histological morphology, classification changes of inflammatory cells and mitogen-activated protein kinase(MAPK) signal pathway in mice with allergic asthma.Method:Forty female C57BL/6 mice were randomly divided into normal control group, model group, dexamethasone group (0.005 g·kg-1) and Da Chengqitang group (19 g·kg-1). Murine allergic asthma model was established by sensitization and nebulization of ovalbumin (OVA). In brief, asthmatic mice were first sensitized by OVA and Al(OH)3 mixture ip on day 0 and day 14, and then nebulized by OVA from day 21 to 27.At the same time, each mouse in the dexamethasone and Da Chengqitang groups were intragastrically administered with 0.2 mL corresponding medicine one hour before the nebulization challenge, while the normal control group was given with the same amount of normal saline. On day 28, pulmonary morphology was detected by htoxylin eosin (HE) staining and inflammatory cells from the brachial alveolar lavage fluid were counted by Diff staining. The expression levels of key proteins in MAPK signaling pathway were detected by Western blot.Result:As compared with the normal control group, the lung indexes were significantly increased in model group (P<0.01), showing obvious inflammatory pathological changes, and serious airway inflammation cells exudation (P<0.01), with a predominant percentage of eosinophils, moreover, the expression levels of phosphorylated p38 MAPK and extracellular signal-regulated kinase 1/2 (ERK1/2) were increased obviously in asthmatic mice. After treatment by Da Chengqitang, lung indexes and pulmonary inflammation were significantly decreased, with an inhibitory rate of 68.4% for lung indexes, and inflammatory pathology of lung tissues was obviously improved and inflammatory cell exudation was alleviated, with the obviously lower levels of phosphorylated p38 MAPK and ERK1/2 protein.Conclusion:Da Chengqitang based on " Pulmonary Intestinal Treatment" can effectively improve lung inflammation in mice with allergic asthma, which may be related to the expression of phosphorylated p38 MAPK and ERK1/2 protein.
Abstract：Objective:To investigate effect of Shaoyaotang on intestinal mucosal immune barrier induced by 2, 4, 6-trinitrobenzene sulphonic acid (TNBS) in rats with ulcerative colitis.Method:Sixty SD rats were randomly divided into normal group, model group, mesalazine group (0.067 mg·kg-1), low, medium and high-dose Shaoyaotang groups (1.8, 3.6, 7.2 g·kg-1). In the TNBS-induced ulcerative colitis model, saline, mesalazine, peony soup were administered by gavage for 7 days. Hematoxylin-eosin(HE) staining was used to detect the histopathological changes of colon tissue. The number of CD4+ T lymphocytes and the expression of secretory immunoglobulin A(SIgA) in intestinal mucosa were detected by immunohistochemistry and Western blot.Result:Compared with normal group, the scores of intestinal mucosal injury and the pathological scores in model group increased significantly (P<0.01). The expressions of CD4+ T lymphocytes and SIgA in the intestinal mucosa of model group decreased significantly (P<0.01). Compared with model group, the scores of colonic mucosal damage and the pathological scores decreased significantly in rats (P<0.01). The expressions of CD4+ T lymphocytes and SIgA in the intestinal mucosa of rats in each group elevated significantly (P<0.05, P<0.01). Compared with mesalazine group and low-dose Shaoyaotang group, the intestinal mucosal injury score and the pathological scores in medium and high-dose Shaoyaotang groups decreased significantly (P<0.01). The expression levels of CD4+ T lymphocytes and SIgA protein in the intestinal mucosa of rats in middle and high doses Shaoyaotang groups increased significantly (P<0.05).Conclusion:Shaoyaotang can reduce the intestinal mucosal damage and protect the intestinal mucosal immune barrier by increasing the number of CD4+ T cells and the expression of SIgA secretion in the intestinal mucosa.
Keywords：Shaoyaotang;2,4,6-trinitrobenzenesulfonic acid;ulcerative colitis;intestinal mucosal immune barrier;secretory immunoglobulin A
Abstract：Objective:To study the effect of Buyang Huanwu Tang on myocardial energy metabolism in rats with diastolic heart failure (DHF) based on adenosine monophosphate (AMP)-activated protein kinase(AMPK)/peroxisome proliferators-activated receptors α (PPARα) signaling pathway, and investigate its mechanism of action.Method:The 48 SD rats were randomly divided into sham operation group and model group. DHF rat model was established by abdominal aorta constriction method. The successfully modeled rats were randomly divided into model group, Buyang Huanwu Tang group (12.72 g·kg-1·d-1), metoprolol tartrate group (0.004 5 g·kg-1·d-1), with corresponding drugs in each group by intragastric administration. The sham operation group and model group were given with equal amount of deionized water, once a day. After 8 weeks of continuous drug intervention, the contents of adenosine monophosphate (AMP), adenosine diphosphate (ADP) and adenosine triphophate (ATP) in peripheral blood of rats were determined by enzyme linked immunosorbent assay (ELISA). The changes of myocardial mitochondrial ultrastructure were detected by electron microscope. The protein expression levels of AMPK, PPARα and peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) in rat myocardium were detected by Western blot.Result:As compared with sham operation group, the contents of AMP and ADP in model group were increased significantly, and ATP content was decreased significantly (P<0.05, P<0.01). As compared with the model group, the AMP content was decreased significantly in Buyang Huanwu Tang group and metoprolol tartrate group (P<0.01), and the content of ADP was decreased while ATP content was increased(P<0.05). As compared with the sham operation group, the number of mitochondria in the model group was decreased, the morphology was abnormal; as compared with the model group, the number of mitochondria in Buyang Huanwu Tang group and metoprolol tartrate group was increased obviously, and the morphology was obviously improved. As compared with the sham operation group, there was no statistically significant difference in the expression of AMPK protein between various groups, but the expression levels of PPARα and PGC-1α protein in the model group were decreased significantly(P<0.05). As compared with the model group, the expression levels of PPARα and PGC-1α protein in Buyang Huanwu Tang group and metoprolol tartrate group were increased significantly (P<0.05).Conclusion:Buyang Huanwu Tang may improve the energy metabolism of the failed heart and delay the progression of heart failure by improving the structure and function of mitochondria, activating AMPK and up-regulating the expression of AMPK/PPARα signaling pathway.
Keywords：Buyang Huanwu Tang;rats with diastolic heart failure;myocardial mitochondria;energy metabolism;adenosine monophosphate(AMP)-activated protein kinase(AMPK)/peroxisome proliferators-activated receptors α(PPARα) signaling pathway
Abstract：Objective:To explore the possible mechanism of Duhuo Jisheng Tang in relieving knee osteoarthritis based on protein kinase R-like endoplasmic reticulum kinase(PERK)/immunoglobulin-binding protein(Bip)signaling pathway.Method:A model of knee osteoarthritis was established by cold stimulation.Rats were randomly divided into blank group, model group, celecoxib group(0.021 g·kg-1), low, medium and high-dose Duhuo Jisheng Tang groups (8.37, 16.72, 33.48 g·kg-1). Blank group and model group were given equal volume of physiological saline.The changes of knee joint diameter were recorded.The pathological changes of rat articular cartilage were observed by hematoxylin-eosin (HE) staining.The expressions of tumor necrosis factor-alpha(TNF-α), interleukin-1β(IL-1β)and hyaluronic acid(HA)in serum were detected by enzyme-linked immunosorbent assay(ELISA). The mRNA and protein expression levels of PERK, Bip and cysteinyl as parates pecific protein-9(Caspase-9)in cartilage were detected by Real-time PCR and Western blot.Result:The knee joint redn ess and the joint diameter of celecoxib group and high-dose Duhuo Jisheng Tang group were improved, and the joint diameter was reduced significantly(P<0.01). The local fractures of knee joint cartilage in the celecoxib group and the high-dose Duhuo Jisheng Tang group were significantly improved.Compared with blank group, the expression levels of TNF-α, IL-1β and HA were increased in model group(P<0.01). The gene and protein expression levels of PERK, Bip and Caspase-9 in cartilage of the model group were increased(P<0.05, P<0.01). Compared with model group, the expressions of TNF-α, IL-1β and HA in serum of celecoxib group and high-dose Duhuo Jisheng Tang group were decreased(P<0.05, P<0.01). The mRNA and protein expression levels of PERK, Bip and Caspase-9 in cartilage of celecoxib group and high-dose Duhuo Jisheng Tang group were decreased(P<0.05, P<0.01).Conclusion:Duhuo Jisheng Tang can alleviate the symptoms of knee osteoarthritis model rats, and its mechanism may be related to the regulation of PERK/Bip signaling pathway in rat cartilage.
Abstract：Objective:To explore the effect of modified Si Junzitang (MSJZT) drug serum on the expression of apoptosis-related molecules of gastric cancer cell SGC-7901 and further its anti-tumor mechanism.Method:A total of 40 SD rats were randomly divided into four groups: low-dose, middle-dose, high-dose MSJZT (0.213, 0.426, 0.853 g·kg-1) groups and normal group (n=10). The treatment groups were administrated through gastric perfusion, and the normal group was given the equivalent volume of normal saline for 10 days. 1.5 h after the last treatment, chloral hydrate peritoneal anesthesia was performed, blood was collected from heart, and different doses of serum were separated to prepare drug-containing serum of low-dose, middle-dose, high-dose MSJZT groups, in order to incubate SGC-7901 gastric cancer cell. Early and late apoptosis rates were detected with flow cytometry. Afterwards, the tumor suppressor gene p53, c-nucleoprotein gene (c-Myc), cysteine-aspartic acid protease-3 (Caspase-3), B-cell lymphoma-2 (Bcl-2) mRNA expressions were confirmed by fluorescence quantitative polymerase chain reaction (Real-time PCR). The protein expressions of p53, c-Myc, Caspase-3, Bcl-2 were detected by immunofluorescence.Result:Compared with the normal group, the high-dose MSJZT group could obviously increase the apoptosis rate to 22.58%(P<0.01). The results of Real-time PCR showed that the middle-dose MSJZT group significantly promoted the mRNA expression of Caspase-3 (P<0.01), the medium-dose group significantly increased the mRNA expressions of p53 and c-Myc (P<0.01), and the high-dose group significantly inhibited the mRNA expression of Bcl-2 (P<0.01). The immunofluorescence results showed that the high-dose MSJZT group could significantly increase the protein expressions of c-Myc, Caspase-3 and p53 (P<0.01), while the protein expression of Bcl-2 was significantly reduced (P<0.05).Conclusion:MSJZT drug serum could exert an anti-tumor effect by inhibiting the expression of the anti-apoptotic protein Bcl-2, and promoting the expressions of pro-apoptotic-related molecules p53, c-Myc, Caspase-3.
Keywords：modified Si Junzitang (MSJZT) drug serum;gastric cancer SGC-7901;apoptosis;tumor suppressor gene p53 (p53);c-nucleoprotein gene (c-Myc);cysteine-aspartic acid protease-3 (Caspase-3);B-cell lymphoma-2 (Bcl-2)
Abstract：Objective:To explore the clinical efficacy of modified Qingqi Huatan Wan in treatment of acute exacerbation of chronic obstructive pulmonary disease (syndrome of phlegm-heat obstructing lung) and investigate its effects on serum tumor necrosis factor-alpha (TNF-α), interleukin-8 (IL-8) and matrix metalloproteinase-9 (MMP-9).Method:Sixty-four patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) were randomly divided into control group (32 cases) and treatment group(32 cases) by random number table.The control group was treated with routine western medicine therapy according to the guidance and disease conditions. Based on treatment in control group, patients in treatment group also received modified Qingqi Huatan Wan. The treatment course was 14 days for both groups.The scores of traditional Chinese medicine (TCM) syndrome, chronic obstructive pulmonary disease (COPD) assessment test (CAT), and modified version of the British Medical Research Council's Respiratory Questionnaire (mMRC), pulmonary function, blood gas analysis indicators, levels of serum TNF-α, IL-8 and MMP-9, clinical efficacy and safety were evaluated and compared once before treatment and 14 d after treatment.Result:The total clinical effective rate was 96.67% in treatment group, higher than 76.67% in control group (χ 2 = 5.192, P<0.05). After treatment, scores of TCM syndrome, CAT and mMRC were reduced in both groups (P<0.05), and the scores in treatment group were all lower than those in control group (P<0.05). Levels of forced expiratory volume in one second (FEV1), percent of FEV1 in predicted value (FEV1%), and ratio of FEV1 to forced vital capacity (FEV1/FVC) were increased in both groups after treatment (P<0.05), and the above levels in treatment group were higher than those in control group (P<0.05). After treatment, oxygen saturation (SaO2) and partial pressure of oxygen (PaO2) were increased in both groups, while partial pressure of carbon dioxide (PaCO2) was decreased (P<0.05), and SaO2 and PaO2 in treatment group were higher than those in control group, while PaCO2 was lower than that in control group (P<0.05). After treatment, serum TNF-α, IL-8 and MMP-9 were decreased in both groups (P<0.05), and the levels in treatment group were lower than those in control group (P<0.05).Conclusion:Modified Qingqi Huatan Wan can control the symptoms safely and ameliorate pulmonary function, reduce the levels of serum TNF-α, IL-8, MMP-9 and inflammation in treatment of AECOPD.
Keywords：acute exacerbation of chronic obstructive pulmonary disease;syndrome of phlegm-heat obstructing lung;modified Qingqi Huatan Wan;interleukin-8 (IL-8);matrix metalloproteinase-9 (MMP-9)
Abstract：Objective:To explore the clinical efficiency of modified Guizhi Gancao Longgu Muli Tang in treating menopausal insomnia.Method:A total of 134 female cases with menopausal insomnia in our hospital from January 2016 to June 2018 were selected as research objectives and randomly divided into observation group (67 cases) and control group(67 cases). Oral zopiclone was provided to control group, and modified Guizhi Gancao Longgu Muli Tang combined with oral zopiclone was provided to observation group. All cases were treated for 4 weeks. The two groups' clinical effects, sleep quality [pittsburgh sleep quality index (PSQI) score], neurotransmitter [glutamate (Glu), γ-aminobutyric acid (GABA), 5-hydroxytryptamine (5-HT), dopamine (DA)] levels before and after treatment, as well as adverse reactions were compared.Result:Observation group's overall effective rate was 92.5%(62/67), which was significantly higher than 80.6%(54/67) of control group (P<0.05). Both groups' scores and total scores of the six dimensions including sleep time, sleep efficiency and sleep quality in PSQI (except for the hypnotic drug dimension) after treatment were significantly lower than those before treatment (P<0.05), and observation group's scores and total scores of each dimension (like sleeping time, sleep efficiency, hypnotic drugs) in PSQI after treatment were significantly lower than those in control group (P<0.05). Compared with those before treatment, both groups' serum Glu, DA levels decreases significantly (P<0.05), while serum GABA, 5-HT levels increased significantly (P<0.05), and observation group showed more significant improvement (P<0.05). Observation group's adverse reaction rate was lower than that of control group, with no statistically significant difference.Conclusion:Modified Guizhi Gancao Longgu Muli Tang can improve sleep quality, alleviate symptoms of insomnia, and correct neurotransmitter metabolic disorder for patients with menopausal insomnia, with an exact curative effect and a higher tolerance of patients.
Keywords：menopausal insomnia;modified Guizhi Gancao Longgu Muli Tang;sleep quality;neurotransmitter;safety;Yin deficiency fire excess type
Abstract：Objective:To explore the effect of modified Xiaokefang on endothelial injury, oxidative stress and biochemical indices of diabetic nephropathy(DN) patients of Qi-Yin deficiency with blood-stasis syndrome.Method:Ninety-four patients with DN admitted to our hospital from September 2017 to June 2018 were randomly divided into two groups. The control group (47 cases) was injected with reduced glutathione by intravenous drip. In addition to the therapy of the control group, the observation group (47 cases) was treated with modified Xiaokefang. Before and after treatment, the symptom scores, fasting blood glucose (FBG), 2 hpostprandial blood glucose (2 hPG), glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD), malondialdehyde (MDA) and interleukin-1 beta (IL-1β), IL-6, hypersensitive C-reactive protein (hs-CRP), tumor necrosis factor-α (TNF-α), tissue blood fibrin dissolve enzyme activation (t-PA), type 1 original organization fibrin lytic enzyme activators inhibitor (PAI-1), serum creatinine (SCr), blood urea nitrogen (BUN), nitric oxide (NO), endothelin-1 (ET-1), 24 h total urinary protein levels were observed, and the efficacy of the two groups were observed.Result:After treatment, the total effective rate of the observation group was 89.36%, which was higher than 74.47%of the control group (Z=3.949, P<0.05). The scores of symptoms in the observation group after treatment were lower than those in the control group (P<0.05). After treatment, FBG, 2 hPG, SCr, BUN, 24 h total urine protein, ET-1, MDA, IL-1β, IL-6, hs-CRP, TNF-α, and PAI-1 levels in the observation group were lower than those in the control group (P<0.05), while NO, SOD, GSH-Px, and t-PA levels were increased (P<0.05).Conclusion:Modified Xiaokefang has a remarkable effect in treatment of diabetic nephropathy patients of Qi-Yin deficiency with blood-stasis syndrome by alleviating clinical symptoms, promoting the recovery of balance of coagulation and fibrinolysis system, inhibiting inflammation and oxidative stress, and protecting endothelial function and renal function.
Abstract：Objective:To observe the short and long term effect of addition and subtraction therapy of Shentong Zhuyutang to myofascial pain syndrome (MPS) with stagnation of blood stasis and to investigate its mechanism of action.Method:One hundred and forty-eight eligible patients were randomly divided into control group (73 cases) and observation group (75 cases) by random number table. Patients in both groups got electroacupuncture treatment. Patients in control group additionally got Yaotong capsules, 4 grains/time, 3 times/day. Patients in observation group additionally got addition and subtraction therapy of Shentong Zhuyutang, 1 dose/day. The treatment was continued for 6 weeks in both groups, and 16 weeks follow-up was recorded. Before treatment, and at the 1st, 2nd, 3rd, 4th, 5th, 6th week after treatment, scores of visual analogue scale (VAS) was graded. Before and after treatment, scores of JOA, Roland-Morris disability questionnaire (RDQ), body damage index assessment scale (PⅡ scale), stagnation of blood stasis, and Pittsburgh sleep quality index (PSQI) were graded. Levels of thromboxane 2 (TXB2), 6-ketone-prostaglandin F1α (6-keto-PGF1α) were detected, and TXB2/6-keto-PGF1α was calculated; in addition, the recurrence was recorded and followed up.Result:By rank sum test, the clinical efficacy in observation group was better than that in control group (Z=1.969, P<0.01). Analysis of variance of repeated measures indicated that, scores of VAS were decreased gradually at the 1st, 2nd, 3rd, 4th, 5th, 6th week in both groups after treatment (Fcontrol=5.801, Fobservation=6.649, P<0.05). Paired t-test indicated that scores of VAS in observation group were lower than those in control group at 2nd, 3rd, 4th, 5th, 6th week(P<0.01). Scores of three dimensions of JOA scale and total score in observation group were higher than those in control group (P<0.01). Scores of RDQ, PⅡ, PSQI, stagnation of blood stasis, whole blood viscosity (high cut and low cut), plasma viscosity, fibrinogen, TXB2, and T/K in observation group were lower than those in control group (P<0.01), and level of 6-keto-PGF1α was higher than that in control group (P<0.01). In addition, the recurrence rate in observation group was 12%, lower than 26.03% in control group (χ2=4.745, P<0.05).Conclusion:Based on acupuncture treatment, addition and subtraction therapy of Shentong Zhuyutang can relieve the pain caused by myofascial pain syndrome (MPS) with stagnation of blood stasis, recover lumbar activity function, reduce rate of recurrence, and ameliorate blood circulation, with anti-inflammatory and analgesia effects.
Keywords：myofascial pain syndrome;stagnation of blood stasis;Shentong Zhuyutang;recurrence;thromboxane 2;6-ketone-prostaglandin F1α
Abstract：Objective:To investigate the protective effect of phenolic compounds 4-hydroxybenzylideneacetone and(HBAc)3, 4-dihydroxybenzylideneacetone (DHBAc) of Osmundae Rhizoma on the systemic inflammatory response syndrome(SIRS) in mice by establishing the mice model of SIRS.Method:BALB/c mice were randomly divided into the normal group, the SIRS model group and the different doses of HBAC and DHBAc group (25, 50, 100 μg·kg-1). Lipopolysaccharides (LPS) was injected intraperitoneally after 7 days of prophylactic administration. After 5 hours of modeling, the anus temperature, respiratory rate, the number of white blood cell(WBC) and platelets(PLT), WBC classification, glycolipid metabolism, inflammatory factor and signal transducing phosphorylated protein of lung were measured.Result:Intraperitoneal injection with LPS (6 mg·kg-1) in mice can significantly reduce the respiratory rate (P<0.05) and the body temperature (P<0.01), decrease the number of WBC, PLT (P<0.01, P<0.01) and blood glucose (P<0.05), and increase the secretion of interleukin-1β (P<0.01). HBAC and DHBAC significantly increased the respiratory rate (P<0.05, P<0.01) and the body temperature (P<0.05, P<0.01) of mice, decreased MONO (P<0.05, P<0.01), and significantly increased the level of blood glucose (P<0.05, P<0.01), and decreased the secretion of interleukin-1β (P<0.01).Conclusion:The SIRS model can be established through intraperitoneal injection of LPS. HBAc and DHBAc have protective effects on endotoxin-induced SIRS in mice, and may exert anti-inflammatory effects through IκB and c-JUN pathways.
Abstract：Objective:To explore the mechanism of Wuzang Wenyang Huayu decoction in improving the cognitive competence and the pharmacological mechanism for neurofibrillary tangles related to cyclin-dependent kinase-5 (CDK-5).Method:The 10 SAMR1 mice were used as normal group, 40 SAMP8 mice were randomly divided into model group, donepezil group (0.4 mg·kg-1·d-1), high and low dose Wuzang Wenyang Huayu decoction groups (5, 1.25 g·kg-1·d-1). Drugs were administered by gastric lavage for 4 continuous weeks. Directional navigation and space exploration ability were evaluated with Morris amaze. Real-time PCR was used to measure the mRNA expression of CDK-5 in brain nerve tissues.Western blot was used to detect the protein expression of CDK-5 and phosphorylation of Tau protein. Meanwhile, neurofibrillary tangles in brain tissue were detected with silver staining method.Result:As compared with normal group, both CDK-5 expression and Tau protein phosphorylation in brain nerve tissues were remarkably increased in model group (P<0.01), with significantly increased neurofibrillary tangles and significantly decreased directional navigation ability and space exploration ability (P<0.01). As compared with model group, CDK-5 expression and Tau protein phosphorylation were lower in Wuzang Wenyang Huayu decoction treated groups(P<0.05, P<0.01), accompanied by decreased number of neurofibrillary tangles as well as improved directional navigation and space exploration ability(P<0.05, P<0.01).Conclusion:Wuzang Wenyang Huayu decoction can markedly improve the cognitive competence of SAMP8 mice, and the mechanism may be related to its inhibition on CDK-5 over-expression, and down-regulation of Tau protein phosphorylation and neurofibrillary tangles in brain tissue.
Abstract：Objective:To investigate the protective effect of compound Longmaining isoprenaline hydrochloride-induced myocardial infarction model and its effect on Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear transcription factor-kappa B (NF-κB) signaling pathway.Method:Forty-eight male SD rats were randomly divided into 6 groups: normal group, model group, compound salvia miltiorrhiza drop pill group(0.072 9 g·kg-1), and low, medium and high-dose compound Longmaining decoction groups(0.36, 0.71, 1.43 g·kg-1). The acute myocardial infarction model was induced through subcutaneous injection with isoproterenol. The pathological changes of myocardial tissue were examined by hematoxylin-eosin (HE) staining. The levels of interleukin-1β(IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α), monocyte chemotaxis protein-1 (MCP-1) and nitrogen (NO) in serum were measured by enzyme linked immunosorbent assay (ELISA). The expression levels of inhibitors of NF-κB kinase subunit-β (IKKβ), NF-κB inhibitor α (IκBα), TLR4, MyD88 and NF-κB p65 were measured by immunohistochemical staining and Western blot.Result:Compared with normal group, the myocardial injury in model group was obvious. The levels of IL-1β, IL-6, TNF-α, MCP-1 and NO in serum increased significantly (P<0.05), the expression level of IκBα decreased significantly (P<0.05), and the expression levels of IKKβ, TLR4, MyD88 and NF-κB p65 increased significantly in myocardial tissue (P<0.05). Compared with model group, low, medium and high-dose compound Longmaining groups could significantly alleviate the degree of myocardial injury in rats, significantly decrease IL-1β, IL-6, TNF-α, MCP-1 and NO levels in the serum (P<0.05), significantly increase the expression level of IκBα (P<0.05), and decreased the expression levels of IKKβ, TLR4, MyD88 and NF-κB p65 (P<0.05).Conclusion:Compound Longmaining plays a protective effect on acute myocardial infarction by regulatingthe expressions of TLR4/MyD88/NF-κB p65 signaling pathway and relevant inflammatory factors.
Abstract：Objective:The present study was designed to investigate the modulating effect of Zhenxin Xingshui Yizhi Fang and its essential oil extract on cognitive deficits in mice.Method:For the purpose of this study 5 months old APP/PS1 double transgenic mice and wild-type C57BL/6JNju were selected as experimental animals. Then APP/PS1 double transgenic mice were randomly divided into model group, essential oil low and high-dose groups (12.13, 48.50 mg·L-1), Zhenxin Xingshui Yizhi Fang group (0.46 g·kg-1). Meanwhile, wild-type C57BL/6JNju mice were used as a normal group. APP/PS1 double transgenic mice were treated with Zhenxin Xingshui Yizhi Fang and its essential oil extract for 22 consecutive days.Mice were subjected to a Morris water maze test and a platform test in order to determine their cognitive effect. Nissl's staining was used to observe pathological changes in brain tissue. Meanwhile, senile plaques (SP) were observed by employing Thioflavin-S staining. The expression of glucose transporter 1 (GLUT1) and insulin receptor substrate-1 (IRS-1) were analyzed using immunohistochemistry techniques. The levels of neurotransmitters such as acetylcholine (ACH), glutamate (GLU) and γ-aminobutyric acid (GABA) in the hippocampus were quantified by enzyme-linked immunosorbent assay(ELISA).Result:The memory function was significantly reduced in model group, and severe brain injury and neuronal apoptosis were also observed in comparison to normal group ( P<0.05, P<0.01). Compared with the model group, Zhenxin Xingshui Yizhi Fang and its essential oil extract improved previous learning and memory impairments (P<0.05, P<0.01), and the levels of SP, IRS-1, GLU, GABA (except essential oil low dose group) were significantly reduced (P<0.01), meanwhile the levels of ACH and GLUT1 were significantly improved ( P<0.05, P<0.01).Conclusion:These results indicate that Zhenxin Xingshui Yizhi Fang and its essential oil extract could ameliorate cognitive deficits and GLUT1 and IRS-1 could be a possible therapeutic target for AD. It may be an interesting approach to the treatment of Alzheimer's disease.
Abstract：Objective:To observe the effect of Prunellae Spica extracts (PS) on the lipid metabolism in Zuker Diabetes Fatty (ZDF) rats based on AMP-activated protein kinase/acetyl CoA carboxylase(AMPK/ACC) signaling pathway.Method:The 32 male ZDF (fa/fa) type 2 diabetic rats were randomly divided into model group, metformin group (180 mg·kg-1·d-1), and low and high-dose PS groups (12.25, 24.5 mg·kg-1·d-1), with 8 in each group. 8 male Zuker Lean (ZL) rats were selected as normal group. Body weight and fasting blood glucose were monitored at the 0th, 4th and 8th weeks after administration. After 8 weeks, abdominal aorta blood was collected, serum was frozen at -20 ℃ by centrifugation, liver tissue was frozen at -80 ℃, fixed with 4%paraformaldehyde and embedded in paraffin. Serum triglyceride (TG), cholesterol (CHO), low density lipoprotein cholesterol (LDL-C) and free fatty acid (FFA) levels were measured by radioimmunoassay. Fat droplets in hepatocytes were measured by oil red O staining. Gene expressions of AMP-activated protein kinase-alpha 2 (AMPKα2), Acetyl CoA carboxylase (ACC) in liver were detected by real-time polymerase chain reaction (Real-time PCR). Protein expressions of p-AMPKα were observed by immuno-histochemical (IHC) method.Result:Compared with the normal group, the T2DM model group showed significant increases in serum levels of TG, CHO, LDL-C, FFA and lipid droplets in hepatocytes. AMPKα2 mRNA expression was decreased, while ACC1 and ACC2 mRNA expressions were increased significantly. p-AMPKα protein expression in liver was decreased significantly (P<0.05, P<0.01). Compared with the model group, low and high-dose Ps groups showed significant decreases in serum levels of TG, CHO, LDL-C, FFA and lipid droplets in hepatocytes in ZDF rats, up-regulation in mRNA expression of AMPKα2, down-regulation in mRNA expressions of ACC1 and ACC2, and up-regulation in protein expression of p-AMPKα (P<0.05, P<0.01).Conclusion:PS can effectively improve liver lipid metabolism in ZDF rats. Its mechanism may be related to the regulation of AMPK/ACC signaling pathway in liver.
Keywords：Prunellae Spica extracts;type 2 diabetes mellitus;lipid metabolism;insulin resistance;AMP-activated protein kinase/acetyl CoA carboxylase(AMPK/ACC)
Abstract：Objective:To investigate the inhibitory effect and mechanism of the extracts from fresh Lycii Fructus(LBL) on hepatoma HepG2 cell-induced cachexia in mouse.Method:The human hepatoma cell line HepG2 was injected into BALB/C mice to establish the cachexia model. Then the LBL was fed to the models respectively in low dose (5 mg·kg-1) or high dose (25 mg·kg-1). After 28 days of continuous feeding, the mice's body weight was detected. The expression levels of creatine kinase (CK), interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were evaluated by enzyme linked immunosorbent assay (ELISA). The muscle degradation and the expression of p38 mitogen-activated protein kinase (p38 MAPK) were detected by immunohistochemistry staining. The expression levels of phospho-p38 mitogen-activated protein kinase (p-p38 MAPK) and nuclear factor-κB (NF-κB) were determined by Western blot.Result:Compared with cachexia group, the loss of body weight and muscle decomposition were significantly inhibited both in the low dose LBL group and high dose LBL group(P<0.05, P<0.01). The level of plasma CK decreased significantly both in the low-dose LBL group and the high-dose LBL group (P<0.01). ELISA tests revealed lower expression levels of IL-1β, IL-6 and TNF-α in both the low-dose LBL group and the high-dose LBL group (P<0.05, P<0.01). Immunohistochemistry staining showed that the expression of p38 MAPK was inhibited both in the low-dose LBL group and the high-dose LBL group (P<0.05). Western blot indicated that the expressions of p-p38 MAPK and NF-κB were inhibited both in the low-dose LBL group and the high-dose LBL group (P<0.01). We found that the high-dose LBL group shows a higher inhibitory capability than the low-dose LBL group.Conclusion:LBL could inhibit the cachexia induced by hepatoma HepG2 cell line in mouse, suggesting LBL could reduce major cytokine and plasma inflammatory factors through p-p38 MAPK pathway.
Abstract：Objective:To evaluate the safety of allantoin extract from Cistanches Herba, so as to provide scientific support for subsequent utilization and development of allantoin extract from Cistanches Herba.Method:According to the national standard procedures and methods in food safety and toxicological evaluation, a series of toxicological studies on allantoin extract from Cistanches Herba were conducted, including genetic toxicity and subacute toxicity tests.Result:In the Ames test, with or without mammalina liver microsomal enzymes (S9), allantoin extract from Cistanches Herba in 40, 200, 1 000, 5 000 μg·dish-1 dose range for four bacteria showed no dose-dependent increase. In the micronucleus test, there was no statistically significant difference among each dose group and the negative control group. In the test of mouse sperm aberration, there was no significant difference in the sperm aberration rate among the allantoin extract group and the negative control group. The results of three genotoxicity tests were all negative, indicating no genotoxicity in allantoin extract. The results of 30 days test showed no death and abnormal clinical sign in rats of control group and each dose group (1.100, 0.550, 0.275 g·kg-1). The body weight, food intake, weekly and total food utilization, weight increment, organ/body ratios, blood biochemical indexes and blood routine indexes had no significant difference among the control group and dose groups. There was no abnormal pathological change in heart, liver, spleen, lung, kidney and testicle of rats in treatment group.Conclusion:Allantoin extract from Cistanches Herba is a non-toxic substance without any genetic toxicity but with a high edible safety. This study provides scientific experimental basis for its safety.
Abstract：Objective:To observe the clinical efficacy of Peiyuan Tongnao capsule combined with " Zhisanzhen" for post-stroke dementia (PSD) and investigate its mechanism.Method:Ninety-eight eligible patients were randomly divided into control group (49 cases) and observation group (49 cases) by random number table. Both groups received donepezil tablet, 10 mg/time, qd, and nimodipine tablet, 30 mg/time, qd. Patients in control group additionally took " Zhisanzhen" , qd, 5 times/week. Based on the treatment in control group, patients in observation group additionally took Peiyuan Tongnao capsule, 3 capsules/time, tid. The treatment course was 12 weeks. Before and at the 4th, 8th and 12th week after treatment, mini-mental state examination (MMSE), montreal cognitive assessment (MoCA), activity of daily living scale (ADL), neuropsychiatric inventory (NPI) and symptoms of traditional Chinese medicine(TCM)were evaluated. Before and after treatment, levels of serum hyperhomocysteinemia (Hcy), hepatocyte growth factor (HGF), oxidized low density lipoprotein (Ox-LDL) and acetylcholinesterase (AchE) were detected.Result:The total clinical effective rate was 93.88% in observation group, better than 77.55% in control group (χ2=5.333, P<0.05). Repeat measurement ANOVA after treatment showed that scores of MoCA, MMSE were gradually increased (Fcontrol=3.947, Fobservation=5.833, P<0.05 for MoCA. Fcontrol=3.876, Fobservation=6.011, P<0.05 for MMSE). At the 8th and 12th after treatment, scores of MMSE in the observation group were all higher than those in control group (P<0.05). At the 12th week after treatment, score of MoCA in observation group was higher than that in control group (P<0.05). Scores of directing force, attention, capacity of calculation in observation group were higher than those in control group after treatment (P<0.05). The changes in scores of MMSE and MoCA in observation group were larger than those in control group after treatment (P<0.05). Scores of ADL, NPI-1, NPI-2 and symptom of TCM in observation group were lower than those in control group (P<0.01). Levels of Hcy, Ox-LDL and AchE were also lower than those in control group, while level of HGF was higher than that in control group (P<0.01).Conclusion:Peiyuan Tongnao capsule combined with " Zhisanzhen" can improve cognitive and behavioral abilities, improve clinical efficacy, ameliorate abnormal mental behavior, relieve clinical symptoms, and regulate levels of Hcy, Ox-LDL, AchE and HGF in the treatment of post-stroke dementia.
Abstract：Objective:To compare the chemical components in raw products and characteristic processed products with porcine cardiac blood of Salviae Miltiorrhizae Radix et Rhizoma from Menghe medical school.Method:The ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF/MS) was performed on an ACQUITY UPLC® C18 column(2.1 mm×100 mm, 1.7 μm), mobile phase was 0.1% formic acid aqueous solution(A)-0.1% formic acid acetonitrile solution(B) for gradient elution(0-1 min, 8%B, 1-1.5 min, 8%-20%B, 1.5-4 min, 20%B, 4-5 min, 20%-60%B, 5-9 min, 60%-70%B, 9-10 min, 70%-95%B, 10-13 min, 95%B). The column temperature was 40 ℃ and the flow rate was 0.3 mL·min-1.Q-TOF/MS with electrospray ionization(ESI) and scanning range of m/z 50-1 200 were applied for analysis under positive and negative ion mode, respectively.All ionic peaks were assigned by comparison of reference substances, mass spectra data, database matching and literature reference, changes of chemical components in Salviae Miltiorrhizae Radix et Rhizoma were investigated by comparing number and area of ionic peaks before and after processing.Result:A total of 59 components were identified from raw products and characteristic processed products, and peak areas of 25 components showed obvious change.However, there were no new compound was found in characteristic processed products.After being processed with porcine cardiac blood, the contents of water-soluble ingredients(protocatechuic aldehyde, salvianolic acid C, F, G), fat-soluble ingredients(tanshinaldehyde, tanshindiol A, tanshinone Ⅰ) and amino acid (L-phenylalanine) in Salviae Miltiorrhizae Radix et Rhizoma were significantly increased.Conclusion:Changes of contents of chemical components in raw products and processed products of Salviae Miltiorrhizae Radix et Rhizoma are remarkable, part of water-soluble ingredients, fat-soluble ingredients and amino acids have quantitative change, which may be related to promoting treatment of Salviae Miltiorrhizae Radix et Rhizoma on cerebral ischemia after being processed with porcine cardiac blood.
Keywords：Menghe medical school;characteristic processing;Salviae Miltiorrhizae Radix et Rhizoma processed with porcine cardiac blood;water-soluble ingredients;fat-soluble ingredients;amino acids;ultra performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF/MS)
Abstract：Objective:To prepare standard decoction of Chuanxiong Rhizoma and conduct its quality analysis.Method:According to the requirements of standard decoction, 15 batches of standard decoction of Chuanxiong Rhizoma from three producing areas were prepared, the HPLC fingerprint was established and the quality analysis was carried out by cluster analysis; under common pattern of fingerprint, the simultaneous determination of four index components(ferulic acid, senkyunolide I, senkyunolide A and ligustilide) was established by HPLC.The transfer rates of main components, dry extract yield, pH value of samples were measured.Result:A total of 15 batches of standard decoctions of Chuanxiong Rhizoma were fingerprinted by Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(edition of 2012A). Twenty-two common peaks were identified, and their similarities were all greater than 0.92, and peak 11, 13, 17, 18, 19 and 20 were identified qualitatively as ferulic acid, senkyunolide I, senkyunolide A, n-butylphthalide, coniferyl ferulate and ligustilide, respectively.The quality overview of standard decoction of Chuanxiong Rhizoma from three producing areas could be distinguished through cluster analysis, which showed that there were differences in quality of Chuanxiong Rhizoma from different producing areas, but the quality was relatively stable in different batches of samples from the same origin.Under common pattern, there were four major components in 15 batches of standard decoction of Chuanxiong Rhizoma, including ferulic acid, senkyunolide I, senkyunolide A and ligustilide.Contents of senkyunolide A(0.176 3-0.249 8 g·L-1) and senkyunolide I (0.065 2-0.103 4 g·L-1) was high in the standard decoction, content of ligustilide(0.040 0-0.089 8 g·L-1) followed, and content of ferulic acid(0.022 0-0.042 3 g·L-1) was the lowest, transfer rates of the above four components were 6.63%-11.82%, 33.32%-55.98%, 1.26%-3.73%and 16.39%-33.05%, respectively.Dry extract yield of the standard decoction was 12.69%to 19.78%, and the pH was 4.54 to 4.82.Conclusion:This study establishes the fingerprint and multi-component determination methods of standard decoctions of Chuanxiong Rhizoma from various producing areas, which is suitable for quality control of this standard decoction.
Abstract：Objective:To compare in vitro dissolution behaviors of active ingredients(gastrodin, parishin A, p-hydroxybenzyl alcohol, parishin B and parishin C) in Gastrodiae Rhizoma powder with different particle size.Method:In vitro dissolution of Gastrodiae Rhizoma powder in different dissolution media(water, artificial gastric juice and artificial intestinal juice) were detected by stirring paddle method.Dissolution of these five components in Gastrodiae Rhizoma powder with different particle size was determined by HPLC, mobile phase was acetonitrile-0.1%phosphoric acid solution for gradient elution, column temperature was set at 40 ℃ and detection wavelength was 220 nm.Result:In water and artificial intestinal juice, the dissolution rates of five active components in three kinds of Gastrodiae Rhizoma ultrafine powders were higher than that of the fine powder and the finest powder; in artificial gastric juice, the dissolution rates of gastrodin and p-hydroxybenzyl alcohol in Gastrodiae Rhizoma ultrafine powders were higher than that of the other powders, and the dissolution rate of parishin A in Gastrodiae Rhizoma ultrafine powders was lower than that of the other powders.Conclusion:An appropriate degree of superfine grinding can promote the dissolution of active ingredients in Gastrodiae Rhizoma, but not as fine as possible.The dissolution medium has an obvious influence on the dissolution behaviors of active components, which provides a reference for screening optical particle size of Gastrodiae Rhizoma powder in clinical application.
Abstract：Objective:To study rhizosphere soil fungal community composition and diversity of Salvia miltiorrhiza, in order to explain the changes of the rhizosphere microenvironment of S. miltiorrhiza, and provide theoretical basis for revealing the reasons for the formation of continuous obstacles.Method:Based on the high-throughput sequencing technology, three kinds of soil samples were collected, namely soil of non-plant (CK), soil for cultivating for one year (Y1) and soil for cultivating for two years (Y2), respectively. According to the sequencing of the internal transcribed space of nuclear ribosomal DNA(ITS) region of fungal rRNA gene, 14 153, 19 024 and 7 622 valid sequences were obtained and annotated as 1 027, 1 095 and 712 varieties of OTUs. The diversity of fungal communities showed a trend of decreasing first and then increasing. Agaricomycetes, Leotiomycetes and Tremellomycetes were the dominant bacterial communities at the class level, their orders in three treatments were Y1>CK>Y2, Y2>Y1>CK, CK>Y2>Y1.At the genus level, Guehomyces was dominant species in CK, and both Y1 and Y2 had a decrease of 96.13%and 90.21%, respectively. Wickerhamomyces had a significantly increase in Y2 (6.64%), 250.59%and 564.00%compared with CK and Y1, respectively.Result:The results of cluster analysis showed that CK and Y1 had a higher similarity at the genus level in soil fungal community composition compared with Y2.Conclusion:Cryptosporiopsis and other pathogenic fungal communities were dominant genuses according to the heatmap analysis in Y2, suggesting a correlation with the incidence of S. miltiorrhiza. The findings provide theoretical basis for the formation of continuous cropping obstacle.
Abstract：Objective:To clone CDS sequence of Dioscorea opposita SUS gene and analyze the protein structure, in order to provide a theoretical basis for the regulation mechanism of SUS gene and the synthesis mechanism of D. opposita polysaccharides.Method:Total RNA in D. opposita was extracted and reverse-transcribed into first strand of cDNA. Specific primers were designed according to an annotated SUS gene sequence obtained from the laboratory D. opposita genome database , and the coding region of the SUS gene was obtained by gene cloning technique and the protein sequence characteristics were analyzed by protein prediction analysis software.Result:A 2 448 bp gene sequence was cloned with a complete open reading frame (ORF). The gene was named DoSUS1.The formula of protein encoded by DoSUS1 gene in D. opposita was C4209H6534N1115O1205S23, and the molecular weight was 9 788.32, the total number of amino acids was 815, the theory isoelectric point(PI) was 6.10, the extinction coefficient was 110 505, the aliphatic index(AI) was 94.15, the instability index was 32.18, and the grand average of hydropathicity(GRAVY) was -0.225.It was a stable and soluble acidic protein. There were phosphorylation sites in the DoSUS1 amino acid sequence, with no transmembrane region and signal peptide. The secondary structure and the tertiary structure showed that DoSUS1 was an α class protein. Functional domain predictions showed that DoSUS1 had sucrose synthesis domain and glycosyl transfer domain. The homology alignment showed that the amino acid sequence of DoSUS1 was more than 80% similar to the amino acid sequence of the aligned monocots. The phylogenetic tree showed that DoSUS1 was closely related to SUS of monocotyledon evolution.Conclusion:The coding sequence of SUS gene was cloned from D. opposita for the first time, and its protein structure was analyzed to lay a foundation for further studying the roles of SUS in the growth and polysaccharide synthesis of D. opposita.
Abstract：Objective:To establish the quality control methods for the standard decoction of Puerariae Thomsonii Radix.Method:According to the preparation principles for traditional Chinese medicine(TCM) standard decoction, 13 batches of Puerariae Thomsonii Radix from different origins were analyzed under the chromatography conditions established in this study and verified with methodology. By referring to Chinese Pharmacopoeia of 2015, puerarin was used as a quantitative indicator to calculate the transfer rate. In this study, the structures of main chromatographic peaks were also identified to clarify the main chemical constituents in the standard decoction.Result:The 13 batches of medicinal herbs were identified as Puerariae Thomsonii Radix, with a recovery rate of 98.0%, and RSD of 1.1%, indicating that the method was accurate and reliable. The transfer rate ranged from 41.4%to 60.0%, and the extraction rate was within the range of 15.7%-34.3%. The corresponding fingerprints were prepared for 13 batches of the standard decoction, and their similarities were all greater than 90.0%. The chemical constituents from Puerariae Thomsonii Radix were identified by mass spectrometry analysis, including citric acid, 4′-O-glucosyl puerarin/daidzein-4′, 7-diglucoside, 3′-hydroxy puerarin/genistein puerarin, puerarin apioside, daidzin puerossid A and daidzein, etc.Conclusion:The 13 batches of Puerariae Thomsonii Radix decoction in different origins had consistent properties with the basic properties of medicinal decoction pieces. The established method of quality evaluation can be used to systematically evaluate the standard decoction, providing reference for quality control of related decoction preparations of Puerariae Thomsonii Radix.
Abstract：Objective:To estimate the overall quality characteristics of Zhenqi Fuzheng granules (ZQFZ), which were composed of Ligustri Lucidi Fructus and Astragali Radix and collected from different manufacturers (their final preparations included two types, contained sugar and sugar free) by established HPLC methods, in order to propose an appropriate quality-control strategy for promoting the quality control specification of ZQFZ.Method:The quantification of the 6 components (rhodioloside, calycosin-7-O-β-D-glucoside, specnuezhenide, ononin, calycosin and astragaloside IV) were performed on a C18 column with two chromatographic systems. Chromatographic system Ⅰ: methanol and water were adopted as mobile phase with gradient elution, the flow rate was 1.0 mL·min-1, and optimum detection waves were at 224, 250 and 275 nm respectively. Chromatographic system Ⅱ: methanol and water (80∶20) were adopted as mobile phase with gradient elution at the flow rate of 1.0 mL·min-1, and the detector parameters were set as follows: the drift tube temperature was 75 ℃, and the carrier gas flow rate was 1.5 L·min-1. Both column temperatures were at 30 ℃. All of the 80 batches of ZQFZ from different manufacturers were determined and analyzed.Result:All of the six markers could be detected in 80 batches of ZQFZ, but their contents were quite different. The results of the one-way ANOVA showed significant differences between manufacturer 4 and other three manufacturers in sugar-containing preparations (P <0.05). And for sugar-free preparations, manufacturer 3 was significant different from other manufacturers (P<0.05). The differences were concentrated in components of rhodioloside and specnuezhenide derived from crude maternal of Ligustri Lucidi Fructus.Conclusion:It is of great significance to increase relevant quality control markers of Ligustri Lucidi Fructus in ZQFZ, such as rhodioloside and specnuezhenide, for standardizing production and improving quality level.
Abstract：Objective:To analyze the change rule of main chemical components in Asarum heterotropoides var. mandshuricum seedling during the growing process.Method:Whole seedling samples(one week and two weeks) and the mature plant(three months) of A. heterotropoides var. mandshuricum were collected and every sample was divided to aerial part(stems and leaves) and underground part(roots). The secondary metabolites were qualitatively identified by HPLC-TOF-MS and the quantitative identification was carried out at the same time with asarinin as index component.Result:A total of 6 known compounds were identified from the underground part of A. heterotropoides var. mandshuricum as α-asarone(1), N-isobutyl-2, 4, 8, 10-dodecatetraenamide(2), 9-methoxyaristolactam Ⅳ(3), asarinin(4), caulesnarinside(6) and chalcononaringenin 2′, 4′-di-O-β-D-glucopyranoside(7), respectively, the peak area values showed that the contents of these compounds increased gradually with the growth time.A total of 4 known compounds were identified from the aerial part of this herb as N-isobutyl-2, 4, 8, 10-dodecatetraenamide(2), caulesauroneside(5), caulesnarinside(6) or chalcononaringenin 2′, 4′-di-O-β-D-glucopyranoside(7) and peonidin 3-caffeoylgentiobioside(8). Asarinin was identified only in the underground part of mature plant, its content was 155.4 μg·g-1.Conclusion:The species and contents of secondary metabolites are quite different in the aerial and underground parts of A. heterotropoides var. mandshuricum.At different growth stages of A. heterotropoides var. mandshuricum seedling, the types and contents of secondary metabolites in the same site are also different, while the contents of main components show an increasing trend with the growth time.
Keywords：Asarum heterotropoides var. mandshuricum;seedling;growth stages;aerial part;underground part;secondary metabolites;asarinin
Abstract：Objective:In order to explore natural active ingredients of Scutellariae Radix and alleviate the pressure of traditional medicinal resources, this research aimmed to screen endophytic fungus strains from Scutellariae Radix which could transform baicalin into some active ingredients.Method:Taking fresh plants of Scutellariae Radix as strain resource, endophytic fungus strains were isolated by potato dextrose agar(PDA) plate separation, Scutellariae Radix powder selective culture and HPLC detection.Result:An endophytic fungus strain R3 was isolated and it can transform baicalin to baicalein and oroxylin A simultaneously in potato dextrose broth(PDB) contained 0.1% baicalin and the conversion rate reached 61.09% in 5 d cultured at 28 ℃ and 150 r·min-1, molar ratio of baicalein and oroxylin A was 3∶5.With the method of morphological analysis, microscopic identification and 18SrDNA sequence analysis, the bacterium was identified as Penicillium sp. R3.Conclusion:Penicillium sp. R3 can transform baicalin to baicalein and oroxylin A with certain research value and application value.
Abstract：As part of systematic research of Corydalis hendersonii, a typical traditional Tibetan herbal medicine with clearing heat, relieving pain, and lowering blood pressure effects, a novel isoquinoline alkaloid, named hendersine G was isolated from the ethanol extract of the whole plant by various chromatographic techniques, including silica gel column, reverse phase column(ODS), Sephadex LH-20, and semi-preparative HPLC. Its structure was elucidated by MS, NMR and other spectroscopic data analysis. Hendersine G can be regarded as a condensation product of a tetrahydroberberine and a succinic acid, however, its absolute configuration has not been determined due to its structural complexity and less obtained amount. This present study provides an inspiration for further exploration of novel molecules from C. hendersonii.
Abstract：Objective:To establish a better near infrared quantitative model for quality control of Glycyrrhizae Radix et Rhizoma of components (moisture, total ash, liquiritin and glycyrrhizic acid) in liquorice, in order to realize rapid detection.Method:The contents of moisture, total ash, liquiritin and glycyrrhizic acid were determined in 97 samples based on the methods set forth in Chinese Pharmacopoeia. Meanwhile, the near infrared spectrum was scanned using near infrared spectroscope. R software was used to screen out the spectral pretreatment and build the quantitative models.Result:The optimum spectral pretreatment method for establishing the near infrared quantitative model of moisture and liquiritin was the first order derivative. For moisture, the correlation coefficients of test and validation were 0.930 0 and 0.929 9, and the root mean square errors were 0.243 2 and 0.203 8, respectively. For liquiritin, the correlation coefficients of test and validation were 0.930 3 and 0.907 6, and the root mean square errors were 0.093 9 and 0.128 9, respectively. The optimum spectral pretreatment method for establishing the near infrared quantitative model of total ash was MSC. The correlation coefficients of test and validation were 0.926 5 and 0.917 7, and the root mean square errors were 0.109 6 and 0.103 7, respectively. The optimum spectral pretreatment method for establishing the near infrared quantitative model of glycyrrhizic acid was SNV. The correlation coefficients of test and validation were 0.918 1 and 0.915 7, and the root mean square errors were 0.274 8 and 0.236 0, respectively.Conclusion:In this study, a better near infrared quantitative models for quality control of components of Glycyrrhizae Radix et Rhizoma were established, with a high accuracy, which laid a foundation for rapid detection of the components in Glycyrrhizae Radix et Rhizoma.
Keywords：Glycyrrhizae Radix et Rhizoma;near infrared;partial least squares;spectral pretreatment;quantitative model
Abstract：Objective:To analyze the medication rules of traditional Chinese medicine (TCM) in treating type 2 diabetes mellitus(T2DM), and explore the mechanism of action.Method:The articlesabout TCM treatment for T2DM in CNKI and Wanfang Med Online were retrieved. The data was entered into Excel 2013 to set up a database, and the factor analysis, association rules and cluster analysis were used to analyze core medication combinations by SPSS Statistics 19.0 and SPSS Modeler 14.1.kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis was performed using BATMAN-TCM to reveal the mechanism of action.Result:A total of 92 prescriptions were included, the high frequency medicines were Astragali Radix.The major properties of TCM were cold and plain.The major flavors of TCM were sweet and bitter. The major meridian tropism of TCM was livermeridian. These high frequency medicines were deficiency-nourishing herbs, and blood circulation-invigorating and stasis-eliminating herbs. Totally 6 core medication combinations were mined through association rules. Seven common factors were extracted from factor analysis. Clustering analysis was used to get 6 kinds of herbs.KEGG pathway analysis revealed 24 biological pathways and 6 diseases relating to T2DM.Conclusion:The most commonly used Astragali Radix for treating T2DM. In clinical, it can be combined with Poria, Salviae Miltiorrhizae Radix et Rhizoma and Rehmanniae Radix for treating T2DM. At the same time, it can be combined with such herbs as deficiency-nourishing herbs, and blood circulation-invigorating and stasis-eliminating herbs. The mechanism of action of high-frequency herbs may be related to the targets of exophosphatase/phosphodiesterase(PDEs), adenosine A1 receptor gene 1(ADORA1), adenosine A2A receptor gene(ADORA2A), neuropetide second receptor(NPY2R), and endothelial carbon monoxide synthase(NOS3) by interfering with PI3K/Akt, starch and sucrose metabolism signaling pathways.
Keywords：type 2 diabetes mellitus(T2DM);medication rule;data mining;traditional Chinese medicine (TCM) prescription;network pharmacology
Abstract：Objective:To explore the key genes and potential therapeutic drugs for osteoarthritis(OA) by bioinformatics.Method:The microarray data GSE55235 was downloaded from the data platform of gene expression omnibus(GEO) and the differentially expressed genes were screened by R language software (3.5.0). Then, the differentially expressed genes were subjected to gene ontology(GO) enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG) signaling pathway analysis with David online database.The protein-protein interaction was analyzed by String 10.5 online database and visual editing was analyzed by Cytoscape v3.6.1 software. Subnetwork module analysis was utilized by MCODE plugin to screen the core genes in the process of OA.Finally, small molecule drugs with potential treatment for OA were analyzed by connectivity map(CMap) database.Result:A total of 556 differentially expressed genes were screened, among which 252 were up-regulated and 304 were down-regulated.These genes were mainly involved in extracellular matrix(ECM) organization, inflammatory response, cell adhesion, immune response, collagen binding, etc.The analysis of KEGG pathway showed that differential genes were mainly involved in ECM-receptor interaction, phosphatidylinositol 3 kinase-protein kinase B(PI3K/Akt) signaling pathway and osteoclast differentiation.Some genes, such as interleukin-6(IL-6), JUN, vascular endothelial growth factor α(VEGFA), FOS, MYC and early growth response gene-1(EGR-1), activating transcription factor-3(ATF-3), playing critical role in the process of OA were identified by protein-protein interaction.Some potential small molecular drugs for the treatment of OA have also been screened, such as lycorine and anisomycin.Conclusion:The selected key genes may be targets for the diagnosis of OA or potential targets for the treatment of OA, and the selected small molecular drugs can be developed as the key drugs for the treatment of OA.
Abstract：Volatile oil is widely distributed in Chinese medicinal materials, which contains rich chemical compositions and has a variety of pharmacological activities such as anti-bacterial, anti-inflammatory, anti-oxidant, anti-tumor, transdermal absorption and antalgic activities, etc. In recent years, the research on the Chinese medicines containing volatile oil is increasing, showing a wide prospect of application in clinical medicine, health care, daily products development, etc. Volatile oil of traditional Chinese medicine has poor solubility, volatility and unstable properties. Factors such as species, harvesting seasons, extraction methods, processing technologies and storage conditions would affect the compositions and contents of volatile oil, thus directly affecting its efficacy, safety and quality stability. Hence, it is especially necessary to control the volatile oil content. The current 2015 edition of the Chinese Pharmacopoeia has made a lot of regulations on the quality control of Chinese medicines containing volatile oil, but generally does not provide the lowest and highest concentrations of volatile oil. As the volatile oil of traditional Chinese medicines has good biological activity and the quality standards are not yet perfect, this study would take the Fist Part of Chinese Pharmacopoeia 2015 Edition as the data source to determine the species of Chinese medicines containing volatile oils through literature research and pharmacopoeia description; the functions, indications, and sources of 196 kinds of traditional Chinese medicines containing volatile oil would be classified and the confusable species would be discriminated by using the Chinese pharmacy knowledge. In this study, the quality-influencing factors would be comprehensively analyzed and the content determination methods of volatile oil would be summarized to offer a reference for the development and application of Chinese medicinal materials containing volatile oil, as well as to lay the foundation for further reinforcing and improving their quality standards.
Abstract：Xanthii Fructus (XF) is one of important drugs for the treatment of sinusitis and headache. It is commonly used in the treatment of diseases relating to immune abnormalities, such as rheumatoid arthritis, acute and chronic rhinitis, allergic rhinitis, and skin diseases, with a high medicinal value. Modern pharmacological studies have showed a wide range of pharmacological effect and a high medicinal value in XF. However, due to long-term or excessive intake, and improper processing of medicinal materials, toxic reactions have often occurred. Toxicological studies have shown that XF poisoning can cause substantial damage to organs, such as the liver, heart and kidney, especially to liver. This paper reviews the pharmacological action, toxic substances and hepatotoxicity mechanism of XF by systematically reviewing and summarizing relevant literatures on XF both at home and abroad. It is concluded that XF has anti-hypertension, anti-allergic, anti-bacterial, anti-inflammatory, analgesic, anti-tumor and lipid-decreasing effects. The toxic components are mainly atractyloside, carboxy atractyloside and 4′-desulphate-atractyloside. The mechanism of hepatotoxicity induced by XF is closely related to lipid peroxidation, bile cholestasis and hepatocyte energy metabolism. Meanwhile, the discovery of novel biomarkers of hepatotoxicity, such as miRNA-122, also provides new ideals for medical research. Toxic traditional Chinese medicine (TCM) is an important part of TCM. Compatibility or processing can reduce or eliminate toxicity and preserve or increase efficacy. At present, there are few reports on the principle of attenuating the production of XF. The author suggests further strengthening the study on the principle of attenuation of XF, giving full play to the unique curative effect of XF and developing its greater medicinal value.
Abstract：Sepsis is a kind of systemic inflammatroy response syndrome (SIRS) induced by severe infection, operation, and trauma, with high mortality rate, treatment cost, and high consumption of medical resources. It has caused a great burden to the medical industry and even the national economy. Therefore, it is urgent to find effective treatment methods for sepsis. At present, the sepsis has been treated with certain drugs pointing at its pathogenesis, such as antibiotics, glucocorticoids, and vasoactive drugs., but the therapeutic effect is not ideal, with many side effects, poor prognosis, and high clinical mortality. Based on the overall macro-dialectical thinking mode, and with the unique effect and low side effect, traditional Chinese medicine (TCM) has attracted the attention from researchers and clinicians around the world for treatment of sepsis. In recent years, some traditional Chinese medicine prescriptions, Chinese patent medicines, single Chinese medicines and active ingredients are increasingly used as new drugs to prevent and treat sepsis. Such treatment methods have been widely recognized and have reduced the mortality and inflammatory indexes of patients to a certain extent, playing an important role in the prevention and treatment of sepsis. In this paper, the actions of nuclear factor kappa B(NF-κB) signal pathway in sepsis as well as the advances in research of NF-κB signal pathway-related proteins in Chinese medicine for sepsis were reviewed.
Keywords：nuclear factor kappa B(NF-κB) signal pathway;sepsis;three syndrome patterns and three treatments;compound Chinese materia;single herb;active ingredient
Abstract：Neural stem cells (NSCs) have the potential of self-renewing and differentiation into neurons, astrocyte, as well as oligodentrocyte. Nowadays, neurogenesis and NSCs transplantation have become new strategies for the treatment of nervous system diseases. However, the disadvantages of NSCs transplantation also limit their progress, including low survival rate, difficulty in differentiation, tumorigenesis and difficulty in penetrating scar tissues. It is a novel issue on how to regulate the proliferation and differentiation of endogenous NSCs in adult neurogenesis.Drug-induced endogenous NSCs proliferation and differentiation is the potential strategy for the regenerative medicine, repairing damaged nerves and treating central nervous system diseases. As a traditional medicine in China, Chinese medicine has the characteristics of multi-target, multi-link and multi-channel treatment, and has unique advantages and broad prospects in promoting nerve regeneration. More and more studies have shown that Chinese herbal medicine could indirectly or directly regulate the proliferation and differentiation of NSCs through extracellular signaling pathways (such as Notch, Wnt/ β-catenin, the Sonic Hedgehog (Shh) pathway), intracellular transcription factors, together with changing the microenvironment (neurotrophic factor BNDF). In this review, we focus on the traditional Chinese medicine formulae, Chinese patent medicine, single medicine, as well as their monomers, which regulate the proliferation and differentiation of NSCs, so as to provide a new strategy for the prevention and treatment of central nervous system diseases by traditional Chinese medicine.
Keywords：neural stem cell;proliferation;differentiation;traditional Chinese medicine;central nervous system diseases
Abstract：Medicinal pair is the cornerstone in compatibility of traditional Chinese medicine prescriptions in clinical application. At present, there are many related studies on Chinese medicine for diabetic nephropathy, but they mainly focus on efficacy observation and prescription summary, lacking in-depth study and summary of the mechanism of action of traditional Chinese medicine(TCM). As compared with prescriptions, Chinese medicinal pair has fewer chemical components, a single form, simple mechanism of action, and fewer interference factors. In recent years, TCM has accumulated rich clinical practice experience in the treatment of diabetic nephropathy. Many researchers pay great attention to the application of medicinal pairs in the compatibility of prescriptions and their action rules. Through the compatibility of medicinal pairs, the optimized combinations can be achieved from various aspects to further improve the therapeutic effect of TCM compound. In order to clarify the ways and targets of Chinese medicinal pairs in treatment of diabetic nephropathy, relevant articles at home and abroad were systemically reviewed in this paper. For the clinical observation and animal experiments of the past 10 years, four types of medicines with " invigorating the spleen and the kidney" , " clearing damp and promoting diuresis" " activating blood and removing stasis" and " securing and astringing" with curative effect for diabetic nephropathy, were mentioned in this article. The composition rules and the progress of experimental researches of these medicinal pairs were analyzed, while the compatibility principles, chemical compositions, and pharmacokinetic changesof the medicinal pairs were summarized to provide some references for their further application, researches and development.
Keywords：diabetic nephropathy;medicinal pair;mechanism of action;research progress