Abstract：Objective:To provide the new quality control means for Alumen by investigating the elemental differences between calcined Alumen and its counterfeit processed products of ammonium alum, and establishing their characteristic chromatogram.Method:The contents of 22 inorganic elements both in calcined Alumen and processed products of ammonium alum were determined by means of inductively coupled plasma (ICP)-optical emission spectrometer-mass spectrometry (ICP-OES/ICP-MS), SPSS 16.0 was used for cluster analysis (CA) while SIMCA-P 13.0 with t-test and Rank-Sum test was used to identify the differential inorganic elements. In addition, the characteristic spectrum of the inorganic elements for calcined Alumen and counterfeit calcined alumen were established.Result:Calcined Alumen had highest contents of K and Al while counterfeit calcined Alumen has highest contents of Al and Fe; Cr, Sr, and Mn contents in calcined Alumen were relatively higher, while Mn, Ti, and Ga contents in processed products of ammonium alum were relatively higher. The content of K in calcined Alumen was about 205 times of that of counterfeit products. On the contrary, the average contents of Fe, Ti, Mn and Ga in counterfeit products of ammonium alum were much higher than those in calcined Alumen, 33, 46, 38, 27 times, respectively. A total of 18 samples were clustered into two categories in CA: calcined Alumen and processed products of ammonium alum. 18 inorganic elements showed significant difference in contents(P<0.05), Be, Mg, Al, K, Ti, V, Mn, Fe, Co, Ni, Cu, Ga, As, Sn, Sb, Hg, Tl, and Pb respectively. Characteristic spectrum of 21 inorganic elements in calcined Alumen and counterfeit calcined Alumen were established.Conclusion:This method can be used for quality control of calcined Alumen.
Keywords：mineral Chinese medicine;calcined Alumen;ammonium alum;ICP-OES;ICP-MS;inorganic elements;quality evaluation;characteristic spectrum
Abstract：Objective:Ammonium alum is a common counterfeit of Alumen, and the processed product of ammonium alum is a common counterfeits of calcined Alumen. This paper aims to establish a method for identifying Alumen, calcined Alumen, ammonium alum and their processed products.Method:The samples were analyzed by scanning electron microscope (SEM) and X ray diffraction (XRD) in this paper.Result:Ammonium alum and Alumen showed obvious changes in morphology after processing. Both Alumen and ammonium alum showed obvious differences in morphology at ×250 and ×1 000 times microscope. Alumen presented irregular fragments, clear edge corners, smooth surface, scattered irregular small particles, occasional holes and longitudinal edges. Ammonium alum presented irregular clumps, blunt edges, not obvious edges and corners, uneven surface, scattered smaller and round-like particles. The difference in morphology was not obvious at ×250 times microscope between Alumen and ammonium alum processed products. While at ×1 000 times, the surface of calcined Alumen was uneven with coarse particles; the surface of counterfeit calcined Alumen was flat, and the coarse particle characteristics were not obvious. XRD can be used to rapidly and accurately identify the primary phase of Alumen, calcined Alumen, ammonium alum and ammonium alum processed products: KAl(SO4)2·12H2O, NH4Al(SO4)2·12H2O, KAl(SO4)2, and NH4Al(SO4)2 respectively, with 2θ angle characteristic value of 23, 12, 22 and 5 respectively for XRD peak.Conclusion:SEM and XRD techniques can be used for the identification of Alumen, calcined Alumen, ammonium alum and their counterfeit products.
Keywords：mineral Chinese medicine;Alumen;calcined Alumen;ammonium alum;scanning electron microscope(SEM);X-ray diffraction(XRD);authentication of Chinese medicine
Abstract：Objective:To evaluate the quality of Limonitum from differentmineral genesis and select its high qualitymineral resources by analyzing itsmineral compositions and content.Method:Themineral compositions and content of different samples were analyzed by polarizing microscope and X-ray diffraction, and goethite content was taken as the evaluation index. SPSS 16.0 statistical software was used for hierarchical cluster analysis of themineral compositions in different samples.Result:Limonite, quartz, illite and mica kaolinite were the main components of Limonitum from leaching disseminated type. Sedimentary type limonite was mainly composed of limonite, quartz and albite. Residual-alteration type limonite was mainly composed of limonite, quartz, diopside and carnallite. The content of goethite in limonite was highest (46.4%) in sedimentary type limonite from Jiangxi province, followed by that in leaching-disseminated type from Jiangsu province (17.9%) and leaching-disseminated type from Shandong province (0.5%). The content of sedimentary goethite was higher than 12.0%and the content of leaching-disseminated type goethite was between 0.5%and 18.0%, with significant difference; and the content of residual-alteration type was below 3.0%. Cluster analysis ofmineral components in different samples showed that 6 samples of leaching-dissemination type were clustered into one group, 3 samples of sedimentation type were clustered into one group, and 2 samples of residual-alteration type were clustered into one group.Conclusion:The quality of sedimentary type Limonitum in Jiangxi is the best. Generally, the quality of sedimentary type is better than that of leaching-disseminated type, and the quality of leaching disseminated type is better than that of residual-alteration type.
Keywords：mineral Chinese medicine;Limonitum;mineral genesis;mineral resources;quality evaluation
Abstract：Objective:To investigate the antidiarrheal effect and the gastrointestinal motility of two kinds of Limonitum with differentmineral genesis, so as to select the optimalmineral genesis, and provide a theoretical basis for clinical use.Method:The selected research samples were leaching type and sedimentary type Limonitum with differentmineral genesis, including powder, water decoction and dregs. In the experiment, castor oil (0.2mL/10 g) was used to induce diarrhea, loperamide (4mg·kg-1) was used for positive control, and then the antidiarrheal activity and the gastrointestinal motility were observed.Result:In the castor oil-induced diarrhea experiment, the diarrhea index of the water decoction (2.5, 5, 10g·kg-1) and the dregs (2.5g·kg-1) of leaching type Limonitum significantly reduced (P<0.05), and so did the degree of diarrhea in mice; the powder of sedimentary type Limonitum, the powder (10g·kg-1) of leaching type limonitum and the dregs (2.5, 5, 10g·kg-1) of the leaching type Limonitum were significant different at the first defecation (P<0.05); the powder (5g·kg-1) and the water decoction (2.5g·kg-1) of leaching type Limonitum, and the total feces weight of the mice were significantly decreased (P<0.05). In the gastrointes motility experiment, the water decoction (5g·kg-1), the dregs (2.5, 5, 10g·kg-1) of the sedimentary, the powder (2.5, 5g·kg-1), the water decoction (2.5, 5, 10g·kg-1) and the dregs (10g·kg-1) of leaching type Limonitum were significantly lower than those of the model group on the intestinal motility, with charcoal as a marker (P<0.05); the dregs (10g·kg-1) of leaching type Limonitum was significantly higher than that of the model group on the content of small intestine contents (P<0.05); the water decoction (10g·kg-1) of sedimentary type limonitum, and the water decoction (2.5, 5g·kg-1) of leaching type Limonitum were significantly lower than those of the model group on gastric residual rate (P<0.05); the powder (5g·kg-1), the dregs (5g·kg-1) of sedimentary type Limonitum were significantly reduced on the water content of large intestine compared with the model group (P<0.05). The 0.003 3, 0.006 7, 0.013 3g·mL-1 of Limonitum can inhibit the contraction of isolated ileum induced by acetylcholine in rabbits except 0.003 3g·mL-1 water decoction of leaching type.Conclusion:Mineral medicine Limonitum has a certain therapeutic effect in castor oil-induced diarrhea. Leaching type limonitum has the best antidiarrheal effect.
Keywords：mineral Chinese medicine;Limonitum;diarrhea;antidiarrheal;castor oil;contraction of ileum
Abstract：Mineral Chinese medicine is an indispensable part in traditional Chinese medicine, with a history of several thousand years in research and utilization of medicinalmineral resources. It is a summary of the medical experience accumulated in numerous trials, observations and practices during the survival process of people of all nationalities, showing distinctive characteristics. At present, the scope ofmineral medicine treatment involves internal medicine, surgery, gynecology, pediatrics, facial features and so on. Themineral Chinese medicine has broad clinical application in many kinds of hemorrhagic diseases, with rich experience in medication and remarkable curative effects. However, there are few papers in modern research, mainly concentrated in the 1970s—1990s. In this paper, the pharmacology and clinical application ofmineral Chinese medicine with hemostatic effects in the last 40 years were reviewed, aiming to provide references for its clinical rational use in bleeding diseases, further development and utilization ofmineral Chinese medicine resources of traditional Chinese medicine, in-depth study of material basis and action mechanism, and further exploration ofmineral Chinese medicine resources. According to the statistics of the major Chinese medicine works, 27 kinds ofmineral drugs have hemostatic effects. The researches on hemostatic pharmacology ofmineral Chinese medicine mainly focus on the analysis ofmineral medicine elements and the effect of related elements on hemostasis, as well as the pharmacological experimental study on hemostasis, but the research is not deep-going and comprehensive. The clinical application ofmineral Chinese medicine for hemostasis mainly includes digestive tract hemorrhage, hemoptysis, epistaxis, gingival hemorrhage, cerebral hemorrhage, post-abortion hemorrhage, metrorrhagia, uterine bleeding, hematochezia and external hemostasis.
Keywords：mineral Chinese medicine;hemostasis;pharmacological research;clinical application;research progress
Abstract：Objective:To observe the effect of Hei Xiaoyaosan on expressions of β-amyloid 1-42 peptide(Aβ1-42), glycogen synthase kinase-3β(GSK-3β), neprilysin(NEP), insulin-degrading enzyme(IDE) in the hippocampus area of Alzheimer's dementia mice.Method:After weighing, 42 APP/PSI bivalent transgenic mice were randomly divided into 4 groups: 10 mice in the model group, 10 mice in the positive drug control group, 11 mice in the high-dose Hei Xiaoyaosan group, and 11 mice in the low-dose Hei Xiaoyaosan group; 10 wild C57BL/6J mice of the same age and strain were used for negative control group. Drugs were administered to mice by gavage once a day for 12 weeks. Then the behavior of all the mice were detected by Morris water maze, the morphological changes in hippocampal neurons were observed by hematoxylineosin(HE) staining, the expressions of Aβ1-42, GSK-3β, NEP and IDE proteins in hippocampus were detected by immunohistochemistry.Result:After 3 months of treatment, compared with negative control groups, the average escaping latency periods prolonged significantly, and the number of cross-platform was decreased significantly in model group (P<0.01), the expressions of Aβ1-42 and GSK-3β proteins in model mice hippocampus were significantly increased (P<0.01), the expressions of NEP and IDE proteins in model mice hippocampus were significantly decreased (P<0.01), suggesting serious damage of hippocampal nerve cells in model group mice according to HE staining; compared with the model group, the escape latency of drug groups were significantly shortened, and the number of crossing platforms was significantly increased (P<0.05), the expressions of Aβ1-42 and GSK-3β proteins in the hippocampus of drug groups were significantly decreased (P<0.01), the expressions of NEP and IDE proteins in the hippocampus of drug groups were significantly increased (P<0.05, P<0.01), suggesting the alleviation in the damage of hippocampal nerve cells in drug groups.Conclusion:Hei Xiaoyaosan can significantly improve the learning and memory abilities of AD mice, which may be related to the reduction of cognitive impairment in AD mice by regulating abnormal deposition and degradating Aβ in the hippocampus.
Abstract：Objective:To investigate the mechanism of Shenling Baizhu San (SLBZS) in treating dextra sulfate sodium (DSS)-induced inflammatory bowel disease (IBD) mice and its relationship with autophagy.Method:SPF BALB/c mice were randomly divided into control group, model group, mesalazine group, low, medium and high-dose SLBZS groups, and autophagy inducer rapamycin group.The IBD mice were fed with 5% DSS in their drinking water for 7 days, and the control mice received only water.SLBZS groups were given SLBZS at doses of 3, 6, 12g·kg-1·d-1, positive group was given mesalazine sustained release granules at the dose of 2g·kg-1·d-1, rapamycin group was given rapamycin at the dose of 4mg·kg-1·d-1, and control mice was given the same volume of normal saline by gavage.The mice weight, stool occult blood in stool, score of disease activity (DAI), pathological examination of intestinal mucosal lesions integral were observed after 7 days. interleukin(IL)-8 and IL-10 in serum were detected by enzyme-linked immuno sorbent assay(ELISA), vascular tissue samples were prepared for the detection of tumor neorosis factor-(TNF-α) and IL-1β, and transmission electron microscope and Western blot were used to detect the formation of autophagosomes and the level of autophagy.Result:The body mass decrease, the colon length, disease activity scoring, and histological scoring of SLBZS group were better than those of DSS group. Compared with control group, the level of IL-10 decreased, while the level of IL-8 increased obviously (P<0.01), IL-1 and TNF-α expressions significantly up-regulated(P<0.01), the formation of autophagosome decreased, and LC3-Ⅱ/Ⅰ level down-regulated. Compared with IBD group, SLBZS group showed increase in formation of autophagosome and LC3-Ⅱ/Ⅰ and IL-10, decrease in IL-8(P<0.05, P<0.01), and down-regulation in IL-1 and TNF-α expressions(P<0.01). These results were observed in mesalazine group and rapamycin group.Conclusion:Shenling Baizhu San can significantly inhibit the IBD by regulating autophagy and suppressing inflammation.
Abstract：Objective:To investigate the effect and mechanism of Xiaoyaosan on lipopolysaccharide(LPS)-induced nerve injury.Method:The 56 rats were randomly divided into control group, sham group, model group, amitriptyline group (10mg·kg-1), fluoxetine group (10mg·kg-1), Xiaoyaosan group high and low-dose (30, 15g·kg-1). The nerve injury model rat were established by LPS injection into lateral ventride, rats were administrated for 14 days by gavage. The levels of brain-derived neurotrophic factor (BDNF) and β-nerve growth factor (β-NGF) in serum were detected by enzyme linked immunosorbent assay(ELISA), and the expressions of BDNF, nerve growth factor (NGF), tropomyosin receptor kinase B (TrkB), tropomyosin receptor kinase A (TrkA), cAMP response element-binding protein (CREB) mRNA in hippocampus and cortex were detected by Real-time PCR.Protoin expression of BDNF, TrkB, CREB, p-CREB, postsynaptic density protein 95 (PSD95), synaptophysin (SYP) in hippocampus and cortex were detected by Western blot.Result:Compared with control group, LPS decreased the level of BDNF and β-NGF in serum(P<0.01), reduced the expression of BDNF, NGF, TrkB, TrkA, CREB mRNA and the expression of BDNF, TrkB, CREB, Phospho-CREB (p-CREB), PSD95, SYP in hippocampus and cortex(P<0.05, P<0.01). Compared with model group, levels of BDNF and β-NGF in serum in Xiaoyaosan high and low-dose group were increased significantly (P<0.05, P<0.01), and expressions of BDNF, NGF, TrkB, TrkA, CREB, PSD95, SYP in hippocampus and cortex were increased significantly (P<0.05, P<0.01).Conclusion:Xiaoyaosan has a certain antagonistic effect on LPS inducednerve injury, which suggests that the effect is related to activate BDNF/NGF-TrkB/TrkA-CREB pathway and upregulated the expression of synaptic protein.
Abstract：Objective:To evaluate the effect of Houpu Sanwu Tang on the postoperative ileus (POI), and observe its underlying mechanisms of action on interstitial cells of cajal (ICC) and inducible nitric oxide synthase(iNOS) regulation of POI.Method:Totally 87 healthy adult male SD rats were randomly divided into sham operation group, saline control group and Houpu Sanwu Tang group at low, medium and high doses. Houpu Sanwu Tang low, middle and high dose groups received orally Houpu Sanwu Tang(2.25, 4.5, 9g·kg-1); Sham operation group (Sham operation) and saline control group (Saline control) received orally normal saline. Surgical procedure was used to induce the postoperative ileus. Changes in intestinal propulsion rate, intestinal mucosal injury and small intestine expression of c-kit and iNOS among these groups were detected.Result:Intestinal propulsion rate was significantly higher in Houpo Sanwu Tang group than that in Saline control group (P<0.01). Rats in Houpo Sanwu Tang group showed reduced intestinal mucosal injury compared with those in Saline control group. C-kit in Houpo Sanwu Tang groups was significantly increased (P<0.01) at 24 h and 48 h, and iNOS in low-dose group and middle-dose group was significantly reduced (P<0.05, P<0.01) at 12 h and 24 h compared with those in the Saline control group.Conclusion:Houpu Sanwu Tang can improve the intestinal propulsion rate and the recovery in POI rats. The mechanisms are related to the inhibition of the generation of iNOS, the alleviation of inflammatory response, and increase of the number of ICC, so as to ensure its normal function, and improve the intestinal dynamic disorder.
Abstract：Objective:To observe the clinical efficacy modified Shenling Baizhu San in treating patients with spleen deficiency wetness type subacute eczema, and its effect on the inflammatory factors and peripheral blood T lymphocyte subsets, so as to provide a theoretical basis for clinical treatment.Method:Totally 176 cases of spleen deficiency wetness type subacute eczema treated at our hospital from Jan. 2016 to Dec. 2017 were selected and randomly divided into control group (88 cases) and observation group (88 cases). Control group was treated with levocetirizine hydrochloride tablets and compound econazole nitrate gel, and observation group was given modified Shenling Baizhu San in addition to therapy of control group for 4 weeks. The clinical efficacy, pruritus score, severity index(EASI)score, dermatology quality of life scales(DQOLS) score and recurrence rate were observed, and inflammatory factor and peripheral blood T lymphocyte subgroup level were detected.Result:After treatment, the pruritus score and EASI score in observation group were lower than those before treatment (P<0.05), and those of observation group were significantly lower than those of control group (P<0.05). However, the DQOLS score was higher than that before treatment, and significantly higher than that of control group (P<0.05). The clinical effective rate of observation group was significantly higher than that of control group (P<0.05). After treatment, the levels of interleukins(IL)-1β, IL-2, IL-4, IL-5 and tumor necrosis factor(TNF)-α in observation group were lower than those before treatment (P<0.05), and the levels of CD4+ and CD4+ /CD8+ were higher than those of before treatment, and significantly higher than those of control group (P<0.05). During the 6-month follow-up, the recurrence rate of observation group was significantly lower than that of control group (P<0.05).Conclusion:Modified Shenling Baizhu San could enhance the clinical efficacy by inhibiting the inflammatory reaction and regulating the immune function of patients.
Abstract：Objective:To investigate the effect of modified Renseng Yangrong Tang combined with chemotherapy on serum tumor markers and Th1/Th2 type cytokines expression in cervical cancer patients.Method:A total of 114 patients with cervical cancer admitted in our hospital from December 2014 to December 2017 were selected as the study subjects. According to the random number table method, the patients were divided into two groups. The control group (57 cases) was treated with conventional chemotherapy. The treatment group (57 cases) was treated with modified Renseng Yangrong Tang in addition to the conventional chemotherapy. Traditional Chinese medicine (TCM) symptom scores, serum tumor markers [carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCCA), carbohydrate antigen 125 (CA125)]and Th1/Th2 type cytokine [interferon gamma-interferon (IFN-γ), interleukin-4 (IL-4), interleukin-2 (IL-2), interleukin-6 (IL-6) levels], clinical efficacy after treatment 3 months, and adverse reaction rate of two groups were observed.Result:There was no significant difference in TCM symptom scores, serum tumor markers and Th1/Th2 cytokines between the two groups before treatment. After treatment, the TCM symptom scores, CEA, SCCA, CA125, IL-6, and IL-4 levels in the treatment group were lower than those in control group and before treatment (P<0.05). The levels of IL-2 and IFN-γ were higher than those in the control group and before treatment (P<0.05). The patients in treatment group were significantly better than those in control group in terms of physical function, mental health, overall health, body pain, social function and vitality (P<0.05), but there was no significant difference in physical role and emotional role. The total effective rate of treatment group (82.46%) was higher than that of control group (64.91%), and the adverse reaction rate (14.04%) was lower than control group (29.82%). The difference was statistically significant (χ2=4.524, 4.150, P<0.05).Conclusion:Modified Renseng Yangrong Tang can significantly relieve clinical symptoms, reduce the level of serum tumor markers, improve cellular immunity, improve the recent therapeutic effect, and reduce adverse reactions in patients with cervical cancer chemotherapy.
Abstract：Objective:To investigate the efficacy of Shensuyin in treating viralmyocarditis (VMC) with syndrome of insufficiency of lung-qi and the effect on levels of Th17 and Treg cells and relevant factors.Method:One hundred-four VMC cases were regaded as object of study and randomly divided into control group and observation group, with 52 cases in each group. Control group was treated with routine therapy by reference to 'Chinese Guidelines for Diagnosis and Treatment of Heart Failure 2014’. In addition to the therapy of control group, observation group on the basis of treatment in the control group with Shensuyin, 1 dose/d, bid. One course of treatment was 8 weeks for both groups. Scores of Shensuyin, serum levels of Troponin I (cTnI) and cardiac free fatty acid binding protein (H-FABP), heart function and total efficacy were compared for both groups. Flow cytometry was used to detect peripheral blood levels of Th17 and Treg cells for the two groups. Serum levels of interleukin (IL)-17, IL-21, IL-10 were detected in both groups.Result:After treatment, scores of syndrome of Yin and Yang deficiency (shortness of breath, fatigue, dull chest pain, bad breath, cough) of observation group were obviously lower than those of control group (P<0.01). Serum levels of H-FABP and cTnI of observation group were remarkably lower, while LVEF and E/A were higher than those of control group after treatment (P<0.01). Total efficacy of observation group was 92.16%, which was higher than 74% of control group (P<0.05). After treatment, peripheral blood levels of Th17 cell and Th17/Treg were obviously lower, while Treg cell was higher thancontrol group (P<0.01). Serum level of IL-17, IL-21 of observation group was remarkably lower, while IL-10 was higher than control group after treatment (P<0.01).Conclusion:In addition to the routine therapy of VMC, the efficacy of Shensuyin has a significant effect in treating VMC with syndrome of lung qi deficiency, and the regulatory effect on levels of Th17 and Treg cells and relevant factors may be one of the effective ways.
Keywords：Shensuyin;viral myocarditis;syndrome of insufficiency of lung-qi;Th17 cell;Treg cell
Abstract：Objective:To observe the efficacy of umbilical moist-heat application of modified Da Chengqitang in treating ventosity after noninvasive ventilation, and the regulatory effect on level of gastrointestinal motility hormone.Method:One hundred and sixty patients were randomly divided into control group and observation group by random number table. Patients in control group got mosapride citrate tablets, 5mg/time, 3 times/days. In addition tothe therapy in control group, patients in observation group was also given mbilical moist-heat application of modified Da Chengqitang, 30min/time, 2 times/day. And a course of treatment was 7 days. Before and after treatment, degree of abdominal distention was measured by a tape and subjective evaluation. Time of recovery of bowel sounds, remission of abdominal distention, exhausting and defecation, and the total time of using respirator, pressure of using respirator, and the total days of using respirator were recorded. And levels of gastrin (GAS), motilin (MTL), vasoactive intestinal peptide (VIP) and somatostatin (SS) were detected.Result:According to the rank sum test, the clinical effect in observation group was superior to that in control group (Z=2.484, P<0.05). Degree of abdominal distention was lower than that in control group (Z=2.456, P<0.05). Difference of abdominal circumference in control group before and after treatment was (14.63±3.76)cm, the observation group is (16.33±4.11)cm, the observation group was more than the control group (P<0.05). The mean time and the total time of using respirator were shorter than those in control group (P<0.05), and pressure was lower than that in control group (P<0.05, P<0.01). And SaO2, PaCO2, GAS, MTL were higher than those in control group, whereas PaCO2, VIP and SS were lower than those in control group (P<0.05, P<0.01).Conclusion:Umbilical moist-heat application of modified Da Chengqitang can relieve abdominal distention and dyspnea, reduce intraperitoneal pressure, improve the recovery of gastrointestinal function, and regulate gastrointestinal hormone level.
Abstract：Objective:To compare the effects between crude and processed Epimedii Folium on the rats of kidney-Yang deficiency and edema, in order to discuss the mechanism.Method:The rat model of kidney-Yang edema was duplicated with hydrocortisone and doxorubicin hydrochloride. At day 1 and day 8 of modeling, the rats were injected with doxorubicin hydrochloride (3.5mg·kg-1) via tail vein, while being intraperitoneally injected with hydrocortisone (3.75mg·kg-1·d-1) for 15days.After modeling, the rats were divided into model group, crude Epimedii Folium group(204.86mg·kg-1)and processed Epimedii Folium group(204.86mg·kg-1), and a normal control group was set up additionally.Rats in each treatment group were given the corresponding drugs, and those in normal and model groups were given the same volume of normal saline by gavage for continuous 14days.At the end of the administration, 24h urine was measured by rat metabolic cage method; 24h urinary protein was detected by ultraviolet spectrophotometry; the content of plasma albumin (ALB) and total serum protein (TP) were measured by an automatic biochemical analyzer; and the content of adenosine monophosphate was measured by enzyme-linked immunosorbent assay(ELISA). Cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), triiodothyronine (T3), thyroxine (T4), serum creatinine (SCr), serum urea(BUN), estradiol (E2), testosterone (T) indicator content were determined by enzyme-linked immunosorbent assay(ELISA). Kidney tissues were collected to make pathological section by htoxylin eosin(HE) staining.Result:Compared with the normal group, all the indicators of appearance, biochemistry and pathological section in the model group indicated kidney Yang deficiency edema in the rats (P<0.05, P<0.01). Compared with the model group, both crude Epimedii Folium group and processed Epimedii Folium group showed a regulatory effect on indicators of appearance and pathological section kidney Yang deficiency edema; and in biochemical indicators, both crude Epimedii Folium group and processed Epimedii Folium group can regulate urine, urine protein, SCr, BUN, cAMP/cGMP, E2, T indicators to varying degrees(P<0.05, P<0.01). Crude Epimedii Folium group showed a significantly regulatory effect on urine, urine protein, SCr, BUN (P<0.01); whereas processed Epimedii Folium group showed a significantly regulatory effect on cAMP/cGMP, E2, T (P<0.01). In addition, crude Epimedii Folium group can also regulate ALB, TP (P<0.05, P<0.01), while processed Epimedii Folium group had no regulatory effect on these two indicators; however, processed Epimedii Folium group could regulate T3, T4 and anal temperature (P<0.05, P<0.01), while crude Epimedii Folium group had no regulatory effect on these two indicators.Conclusion:Both crude Epimedii Folium group and processed Epimedii Folium group have certain effect in treating kidney Yang deficiency edema, and the possible mechanism is to alleviate glomerular podocyte injury induced by doxorubicin. Their emphasis in the treatment of kidney Yang deficiency edema is different. Crude Epimedii Folium has a cold property, and can treat kidney Yang deficiency edema by strengthening renal excretion function of rats with kidney Yang deficiency edema. Processed Epimedii Folium is processed with sheep oil to produce icariin and other substances, and its property changed from cold to warm, with a focus on alleviating the kidney Yang deficiency status of rats with kidney Yang deficiency edema.
Keywords：processed Epimedii Folium;crude Epimedii Folium;kidney Yang deficiency edema;electron microscopy observation
Abstract：Objective:To observe the effect of Weichang'an on the tumor growth and lymph node metastasis, and RUN and FYVE domain-containing protein 3(RUFY3), Zinc finger protein 1(SNAI1), vascular endothelial growth factor(VEGF), epithelial-mesenchymal transition(EMT)-related proteins in nude mice with subcutaneous xenograft tumor human gastric carcinoma MKN45, so as to discuss the mechanism of Weichang'an on MKN45 human gastric metastasis.Method:The nude mice model of human gastric carcinoma MKN45 cells was established and randomly divided into normal saline group (0.5mL/mouse), Weichang'an granule group (3.54g·kg-1) and Weichang'an decoction group (35.49g·kg-1). The tumor weight and volume of axillary lymph nodes in each group were observed. The morphology of lymph nodes in each group was detected by hematoxylin-eosin (HE) staining. The expressions of related proteins were detected by immunohistochemistry, including RUFY3, SNAI1, VEGF, E-cadherin, N-cadherin, Vimentin.Result:Compared with the normal saline group, the tumor weight and volume of axillary lymph node were decreased (P<0.01). HE staining revealed metastatic tumor cells in all lymph nodes. The positive expression rates of RUFY3, SNAI1, VEGF, N-cadherin, Vimentin in the Weichang'an granule group and Weichang'an decoction group were lower than those in normal saline group (P<0.01), while the positive expression of E-cadherin increased significantly (P<0.01).Conclusion:Both Weichang'an granule and Weichang'an decoction can inhibit the tumor growth and metastasis of axillary lymph nodes, obviously down-regulate the expressions of RUFY3, SNAI1, VEGF, N-cadherin, Vimentin and up-regulate the expression of E-cadherin in human gastric MKN45 subcutaneous transplanted tumor in nude mice. This suggested that Weichang'an may inhibit the tumor metastasis through regulation expressions of RUFY3, SNAI1, VEGF and EMT-related proteins.
Keywords：Weichang'an granule;human gastric cancer cell;subcutaneous transplanted tumor;lymph node metastasis;RUN and FYVE domain-containing protein 3(RUFY3);Zinc finger protein1(SNAI1);vascular endothelial growth factor(VEGF);epithelial-mesenchymal transition(EMT)
Abstract：Objective:To investigate effect of Yanghe Huayan Tang and phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 in intervening expressions of human epidermal growth factor receptor human epidermal growth factor receptor-2(HER-2), PI3K and phosphorylated serine/threonine kinase (p-Akt)in PI3K/Akt pathway of breast cancer cell line SK-BR-3 (HER-2 high expression) in vitro, and study intervention mechanism of Yanghe Huayan Tang.Method:Yanghe Huayan Tang intestinal absorption fluid was prepared, breast cancer cell strain SK-BR-3 was divided into blank group, Yanghe Huayan Tang group, LY294002 group, LY294002+ Yanghe Huayan Tang group. The concentration was respectively 125 g·L-1 for Yanghe Huayan Tang, 0.05 g·L-1 for LY294002, 125 g·L-1 for Yanghe Huayan Tang+ LY294002.The expressions of HER-2, PI3K and p-Akt protein were detected by immunohistochemistry and Western blot after 24 h. The most appropriate and effective concentration was obtained through extensive experiments. The expressions of HER-2, PI3K and p-Akt were detected by reverse transcription polymerase chain reaction (RT-PCR).Result:Compared with blank group, LY294002 group, and LY294002+ Yanghe Huayan Tang group could inhibit protein and mRNA expressions of HER-2, PI3K, p-Akt in breast cancer cell lines(P<0.05). Yanghe Huayan Tang group could inhibit expressions of HER-2, p-Akt and mRNA (P<0.05) in breast cancer cell lines, but inhibitory effect on PI3K was not obvious; combination with LY294002 could further inhibit PI3K expression compared with single administration with LY294002 (P<0.05). Compared with LY294002 group, LY294002+ Yanghe Huayan Tang group had a more obvious inhibitory effect. The combined administration of Yanghe Huayan Tang and LY294002 could inhibit protein and mRNA expressions of HER-2, PI3K, p-Akt significantly (P<0.05).Conclusion:Yanghe Huayan Tang can inhibit angiogenesis and invasion of breast cancer. Its main mechanism is expressed by intervening PI3K/Akt pathway, reducing expressions of HER-2, PI3K and p-Akt in pathway.
Keywords：Yanghe Huayan Tang;human epidermal growth factor receptor-2(HER-2) high-expression breast cancer;phosphatidylinositol 3-kinase (PI3K) inhibitor;vasculogenesis and invasion;PI3K/serine/threonine kinase (Akt) pathway
Abstract：Objective:To observe effect of Tangshenping capsule on lipopolysaccharide (LPS)-stimulated podocytes apoptosis under high glucose conditions of podocyte in rats with diabetic nephropathy (DN), and discuss possible mechanism.Method:With cultured rat podocytes as object of study, the podocytes model was established with high glucose and LPS, and divided into 7 groups: control group, high glucose group, high glucose plus LPS group, irbesartan group, and low, moderate and high-dose Tangshenping capsule groups.Protein and mRNA expressions of CD2-associated protein (CD2AP), nephrin, phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and B-cell lymphoma-2 associated death protein (Bad protein) of podocyte in each group were detected by Western blot and reverse transcription polymerase chain reaction (RT-PCR).Result:Compared with control group, the protein and mRNA expressions of podocyte CD2AP, nephrin, PI3K, Akt decreased definitely (P<0.05, P<0.01), while Bad protein and mRNA expressions increased definitely (P<0.05, P<0.01). Compared with high glucose plus LPS group, the protein and mRNA expressions of podocyte CD2AP, PI3K, Akt increased definitely (P<0.05, P<0.01), whereas Bad protein and mRNA expressions decreased definitely (P<0.05, P<0.01) in irbesartan group. The protein and mRNA expressions of podocyte nephrin, CD2AP, PI3K, Akt increased definitely (P<0.05, P<0.01), and Bad protein and mRNA expressions decreased definitely (P<0.05, P<0.01) in three Tangshenping capsule groups.Conclusion:Tangshenping capsule can maintain a stable protein expression of Slit diaphragm (SD) in podocyte, inhibit podocyte apoptosis by activating PI3K/Akt signaling pathway. This is one of mechanisms for preventing and treating diabetic nephropathy.
Keywords：diabetic nephropathy;podocyte;Tangshenping capsule;phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway
Abstract：Objective:To observe the effect of phosphoinositide-3-kinase (PI3K)-mediated nuclear factor E2-related factor 2 (Nrf2) phosphorylated activation in quercetin (Quer) in prohibiting clivorine (Cliv)-induced cytotoxicity in L-02 cells.Method:Human normal liver L-02 cells were pre-incubated with Quer (1, 10, 25, 50 μmol·L-1) for 15min, and then incubated with Cliv (50 μmol·L-1) for 48 h. The cell viability was evaluated by thiazolam blue(MTT) assay. Cellular reactive oxygen species (ROS) and reduced glutathione (GSH) were detected. The transcriptional activation of Nrf2 was measured by dual-luciferase reporter assay. The phosphorylation of Nrf2 and PI3K was measured by Western blot, and downstream genes, including heme oxygenase 1 (Hmox1), NADPH: quinone oxidoreductase 1 (Nqo1), catalytic subunit of glutamate-cysteine ligase (Gclc), modifier subunit of glutamate-cysteine ligase (Gclm), were measured by Real-time PCR.Result:Quer (10, 25, 50 μmol·L-1) reversed Cliv-induced decreased cell viability (P<0.05), GSH (P<0.05) and ROS levels (P<0.01). Quer induced phosphorylation of PI3K (P<0.05). Quer induced phosphorylation and subsequent transcriptional activation of Nrf2 (P<0.05), and enhanced mRNA expressions of Nrf2 downstream genes (P<0.05). The increased Hmox1 mRNA expression was reduced by PI3K inhibitor LY294002 (LY) (P<0.01).Conclusion:Quer alleviates Cliv-induced hepatotoxicity by inducing Nrf2 phosphorylated activation, and PI3K plays an important role in regulating Quer-induced Nrf2 phosphorylated activation.
Abstract：Objective:To observe the effect of diosgenin on aplastic anemia (AA) mice and peroxisome proliferator activated receptor γ (PPARγ), CCAAT-enhancer binding protein α (C/EBPα), Adiponectin, Leptin, in order to discuss the potential mechanism of bone marrow mesenchymal stem cells in the process of adipemia.Method:BALB/c mice were randomly divided into control group and model group. The model group was established by 60Coy irradiation combined with tail vein infusion with lymphatic suspension cells of DBA/2 mice. After successful evaluation of the model, the mice were randomly divided into 6groups: model group, low, medium and high-dose diosgenin groups (37.44, 74.88, 149.76mg·kg-1·d-1), cyclosporine group (23.5mg·kg-1·d-1), and tripterygium glycoside group (9.36mg·kg-1·d-1), and given corresponding drugs by gavage for 14days. After the intervention, the peripheral blood of mice in each group was detected, and bone marrow smears were collected to evaluate the proliferation of bone marrow. Bone marrow mesenchymal stem cells (BMMSCs) were isolated and cultured by adherent method and induced by adipogenesis. ThemRNA and protein expressions of PPARγ, C/EBPα, Adiponectin, Leptin in BMMSCs were detected by quantitative real-time fluorescent quantitative polymerase chain reaction(Real-time PCR) and Western blot.Result:The white blood cell (WBC), hemoglobin (HGB) and blood platelet (PLT) in peripheral blood of model group were significantly lower than those of normal group (P<0.05). The hemogram of each treatment group was higher than that of model group to varying degrees. The middle-dose Dioscorea nipponica group had the most obvious hemogram, with statistically significant differences compared with the model group (P<0.05). Compared with the control group, the mRNA and protein expressions of PPARγ, C/EBPα, Adiponectin and Leptin in BMMSCs of the model group increased significantly (P<0.05). Compared with the model group, the above indexes in the treatment groups were all decreased, the mRNA and protein expressions of PPARγ, C/EBPα, Adiponectin and Leptin in the middle-dose group diosgenin decreased obviously, which was better than those of Tripterygium glycoside group (P<0.05).Conclusion:Diosgenin can promote the recovery of peripheral blood in aplastic anemia mice and improve the hematopoiesis of bone marrow. Diosgenin can reduce the expressions of PPARγ and C/EBPα, the formation of adipocytes and the secretion of Adiponectin and Leptin in adipocytes, and effectively inhibit the process of adipose tissue derived from bone BMMSCs.
Abstract：Objective:To explore the effect of Tongbiantang on protein kinase A(PKA) and mitogen-activated protein kinase(MAPK) signal pathway in colon tissue of slow transit constipation(STC) rats and its related mechanism.Method:Eighty SD rats were randomly divided into blank group and model group, 20 rats in blank group, 60 rats in model group, half male and half female; blank group was fed with common diet, model group was fed with compound phenylethylpiperidine, after 120days of modeling, 10 rats in blank group and 20 rats in model group were randomly selected, and 2 rats were determined. Four-hour stool volume, water content and small intestinal charcoal powder propelling rate were observed to observe the number of stool particles retained in colon and evaluate the success of STC rat modeling. After 1 week of drug withdrawal, 40 rats in model group were randomly divided into model group(33g·kg-1), Tongbiantang group, Tongbiantang+ H89group (PKA signaling pathway blocker, 5mg·kg-1), Tongbiantang+ U0126group (MPKA signaling pathway blocker, 0.1mg·kg-1) each. After 4 weeks of intervention with Tongbiantang, the amount of stool excretion, water content and small intestinal charcoal powder propelling rate were measured in 10 rats, and the number of stool grains in colon was observed. The protein content andmRNA expression in aquaporins 3(AQP3), AQP4, PKA and MAPKs signaling pathways in colon was determined by immunohistochemical staining (IHC), Western blot and Real-time fluorescence quantitative PCR (Real-time PCR).Result:Compared with the blank group, the 24-hour stool volume, fecal water content, small intestinal charcoal propelling rate and the number of fecal particles in colon of rats in the model group were significantly decreased (P<0.01); compared with the model group, the fecal volume, water content and small intestinal charcoal propelling rate of Tongbiantang group were increased, the number of fecal particles in colon decreased (P<0.01), AQP3 and AQP4 were significantly decreased (P<0.01). Compared with the Tongbiantang group, the expressions of AQP3, AQP4, PKA andmRNA in the Tongbiantang+ H89group and the Tongbiantang+ U0126group were significantly lower (P<0.01); compared with the Tongbiantang+ H89group, the stool volume, water content, small intestine charcoal propelling rate and the number of stool particles in colon, AQP3, AQP4, PKA, MPKAs protein table were significantly lower (P<0.01). There was no significant difference between the amount ofmRNA and the content of protein.Conclusion:Tongbiantang can inhibit the PKA and MPKA signal pathways, thus down-regulate the expression of AQP3 and AQP4, increase intestinal peristalsis and intestinal water, and effectively treat STC.
Keywords：Tongbiantang;slow transit constipation;protein kinase A(PKA)/mitogen-activated protein kinase(MAPK) signal pathways;aquaporins 3, 4
Abstract：Objective:To investigate the effect of dodder total flavone on polycystic ovary syndrome (PCOS) rat models induced by dehydroepiandrosterone (DHEA) combined with human chorionic gonadotropin (HCG).Method:Except the blank group, the remaining rats were injected with DHEA 0.06mg·g-1 in the morning on the nucha and 1.5 U HCG in the afternoon for 21 consecutive days. On the 16th day after the modeling, the vaginal smear was performed to monitor the estrus cycle. Sixty rats with successful modeling were selected and divided into model group, dacin-35 group, and high, middle and low-dose dodder total flavonoids groups, with 10 rats in each group. On the day of grouping, drugs were given respectively to the drug treatment groups, and the blank group and the model group were given equal volume solvent. The drugs were given continuously for 21 days. Blood was collected from abdominal aorta 2 h later after the final administration, serum levels of testosterone (T), estrogen (E2), luteinizing hormone (LH) and follicle stimulating hormone(FSH) were measured; rats were put to death, and the ovaries at the same part of each rat were fixed in 10% formalin solution. htoxylin eosin (HE) staining was performed, and the morphological changes in the ovaries were observed by light microscopy; the same part of the ovary was taken, and androgen receptor (AR), B-cell lymphoma-2(Bcl-2) and Bcl-2-associated X protein(Bax) were measured in the ovary by immunohistochemistry to observe the expressions of relevant proteins in the ovary; the hypothalamus and pituitary were taken, and the expressions of gonadotropin-releasing hormone (GnRH) in hypothalamus and gonadorelin releasing hormone receptor (GnRHR) in pituitary were detected by immunohistochemistry.Result:PCOS rat model was successfully replicated. Serum levels of T, E2 and LH were significantly reduced, and FSH level of PCOS was significantly increased in each drug treatment group (P<0.01). The pituitary GnRHR level was increased to different degrees (P<0.05, P<0.01); AR and Bax levels of the ovary were decreased to different degrees (P<0.05, P<0.01); hypothalamus GnRH level was significantly decreased, and pathological changes in ovarian tissue of PCOS rats were obvious (P<0.01) in each drug treatment group.Conclusion:The PCOS rat model was successfully replicated. Total flavonoids of dodder may play a protective role in PCOS model rats by regulating the secretion of androgen, inhibiting the expression of ovarian apoptotic protein and impacting the hypothalamic-pituitary-ovary axis pathway.
Keywords：polycystic ovary syndrome;apoptosis proteins;dodder total flavone;biochemical indicators
Abstract：Objective:In this paper, the effect of microemulsion in Chuanqi ophthalmic microemulsion in situ gel was investigated.Method:The effect of microemulsion was confirmed by the parallel comparison between the Chuanqi microemulsion in situ gel and normal in situ gel, including study of pharmaceutical characterization and tissue distribution.Result:The average particle sizes of Chuanqi microemulsion in situ gel and normal in situ gel were (38.20±0.13) nm and (985±37) nm, respectively.Microemulsion could maintain the properties of nanocarrier in a microemulsion in situ gel composite system.The result of tissue distribution study showed that only ligustilide could be detected.This was related to the nature of these three indicator components(ligustrazine, ligustilide and astragaloside A). The common logarithm of oil and water partition coefficient of ligustilide(lgP) was 2.87, which was consistent with the range of lgP of ideal ophthalmic drugs(lgP=2.0-3.0). The ligustilide from Chuanqi microemulsion in situ gel could be detected in the cornea, vitreous body and retina, and this compound from normal in situ gel could only be detected in the cornea with low content.At the same time, microemulsion could increase the content of ligustilide in corneal tissues.Conclusion:The characteristics of microemulsion nanocarriers can increase the solubility of ligustilide, compared with normal in situ gel, it can be better distributed in the tears outside the corneal, it reaches the cornea with a higher concentration, and forms a corneal concentration gradient, and ligustilide is transported from the anterior ocular region to the posterior ocular region through the transocular barrier.
Keywords：microemulsion;tissue distribution;cornea;ligustilide;particle size;Chuanqi microemulsion in situ gel;normal in situ gel
Abstract：Objective:To establish a method for determining the content of total polysaccharides in decoction pieces of Polyporus, analyze the content of total polysaccharides in samples with different sources and grades.Method:The relative molecular weight and the polydispersity index of polysaccharides in decoction pieces of Polyporus were measured by a high performance gel chromatography coupled with a multi-angle laser light scattering and refractive index system.Dextran with similar molecular weight as polysaccharides was selected as the reference substance.Orthogonal experiment and single factor tests were used to optimize the pretreatment conditions for the determination of total polysaccharides in Polyporus.Polysaccharides in Polyporus with different areas and grades were determined by anthrone-sulfuric acid colorimetric method at 630nm.Result:The linearity, stability, precision, repeatability and recovery rate of the established method all reached the standards, respectively.The content of total polysaccharides in samples from different areas ranged from 0.87% to 1.39%.The content of total polysaccharides in samples with different grades was 1.40% for first-grade pieces, 1.21% for second-grade pieces, and 1.03% for third-grade pieces.Conclusion:The established method is simple, accurate and reproducible, and it can be used for the determination of polysaccharides in decoction pieces of Polyporus.The content of polysaccharides in samples from different origins varies greatly.The content of polysaccharides in samples with different grades shows a certain regularity.The content of polysaccharides is the highest in the first-grade pieces, followed by the content in the second-grade, and the lowest in the third-grade.The results can provide a reference for formulating limits for the content of total polysaccharides and the grade standard of decoction pieces of Polyporus.
Keywords：Polyporus;total polysaccharides;dextran;anthrone-sulfuric acid colorimetry;relative molecular weight;high performance gel chromatography;commercial grade
Abstract：Realgar is toxic and belongs to drug of poison attack, with anti-cancer, anti-pest, dry wet and expectorant effects. Ancient doctors often used realgar to treat carbuncle, boil, abdominal pain and other diseases. Modern doctors use it to treat malignant tumors and blood diseases. The toxicity of realgar results in a small range of safety in its drug use. Modern scholars combine traditional Chinese medicine with nano-technology means to grind realgar into nano-realgar. Compared with realgar, nano-realgar has an improved bioavailability and reduced toxicity in vivo. Modern pharmacological researches use nano-realgar to interven lung cancer cells, skin cancer cells, cervical cancer cells, ovarian cancer cells and leukemia cells in experiments, which confirmed that nano-realgar has effects in inhibiting cell proliferation, inducing apoptosis and cell differentiation, inhibiting nucleic acid synthesis and angiogenesis, but with antiviral, sterilization of analgesic effects. Modern toxicological studies have been conducted in mice through intragastric treatment with different concentrations of nano-realgar preparation. The symptoms and signs of mice and various auxiliary examination indexes were recorded at different time periods. It was concluded that nano-realgar has regular effects on blood biochemical indexes at safe doses. In clinical trials, nano-realgar was applied to treat damaged wound surface of malignant tumor. It was found that nano-realgar can promote healing of wound surface of tumor, alleviate pain, relieve clinical symptoms, such as bleeding, purulence and odor, and improve quality of life of patients. In addition, with a simple usage and good patient compliance, administration with nano-realgar requires no hospitalization, can save medical resources, and is worthy of clinical promotion and application.
Abstract：Diabetes mellitus (DM) is a metabolic disease characterized by hyperglycemia, with a high incidence and many complications. It has become an increasingly serious public health problem in the world, and has seriously affected the quality of life. Phosphatidylinositol-3-kinases (PI3K)/protein kinase B (Akt) signaling pathway is the main pathway of insulin signal transmission and the main signal channel for regulating blood glucose. The abnormal signal molecule of PI3K/Akt may cause abnormal signal transduction pathway, so as to impact the proliferation, apoptosis, metastasis and invasion of the corresponding tissues and organs, and lead to the occurrence of disease. Study of PI3K/Akt signal channel has a positive significance for investigating whether traditional Chinese medicine(TCM) has a definite and stable hypoglycemic effect. Currently, there are many TCM and Western medicines to treat diabetes, however, most drugs, especially Western medicines, have a relatively poor effect in controlling complications. To understand the progress of TCM in treatment of diabetes, in expectation of better studying the comprehensive therapeutic effect and mechanism of TCM on diabetes, and further developing the multi-target, multi-way and multi-channel advantages and features of TCM in the treatment of diabetes, this paper focuses on a systematic analysis on the progress of in vivo and in vitro studies on DM based on PI3K/Akt signaling channel in recent years, including the effect of the signaling channel on insulin secretion, the three main target organs of insulin (liver, skeletal muscle and fat), and its effect on the four main complications of diabetes (brain, kidney, heart, testis), and also provides certain ideas and guidance for the study of hypoglycemic mechanism of TCM monomer, TCM and compound medicine.
Keywords：phosphatidylinositol-3-kinases(PI3K)/protein kinase B(Akt)pathway;diabetes mellitus;traditional Chinese medicine;target organ;complication;insulin
Abstract：Cinobufacini is an extract from skin of Bufo gargarizans, and composed of bufadienolides, peptides, alkaloids and cholesterol. With the deepening of research of its pharmacological effect, it has been proved that cinobufacini has important application value in anti-inflammation, anti-virus, anti-tumor, as well as swelling and pain relief. In recent years, more attention has been given to its anti-tumor activity. The molecular pharmacology studies showed that cinobufacini plays an important role on anti-tumor by enhancing the immunity, inhibiting the proliferation of tumor cells, inducing the apoptosis of tumor cells, reversing multidrug resistance of tumor cells, and inhibiting the tumor angiogenesis. Both intrinsic pathway mediated by mitochondria and extrinsic pathway mediated by death receptor are involved in cinobufacini-induced apoptosis. Clinical studies have shown that cinobufacini is broadly used as an anti-tumor traditional Chinese medicine, due to its advantages of low toxicity, less side effects, and wide anticancer spectrum. It is broadly used in the treatment of liver cancer, gastric carcinoma, lung cancer, colon cancer, esophageal carcinoma, carcinoma of gallbladder, and non-Hodgkinlymphoma. Either single or combined application plays an active role in improving the efficacy and reducing side effects. To promote the research of anti-tumor activity of cinobufacini, and expand its application prospects, this review focuses on anti-tumor mechanism and clinical application of cinobufacini.
Abstract：Objective:To establish the quality control methods for the standard decoction of Zingiberis Rhizoma.Method:DNA barcode primitives were identified for the medicinal materials from different origins; according to the standard of Chinese herbal medicine decoction preparation principle, the identified Zingiberis Rhizoma was prepared into standard decoction for analysis. Meanwhile, the extraction method and analysis method were validated from methodologies, and the transfer rate of 6-gingerol as well as the extraction rate of standard decoction of Zingiberis Rhizoma were calculated. In addition, the quality standard of standard decoction of Zingiberis Rhizoma was also established based. The structures of main chromatographic peaks were identified by ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) to clarify the main chemical constituents in the standard decoction of Zingiberis Rhizoma.Result:All the samples were identified as Zingiberis Rhizoma. Under the conditions established in this paper, the standard curve of 6-gingerol was Y=661.56X+ 2.493 3(r=0.999 3), and the RSD was 0.5%in precision test, indicating that the instrument precision was good. The repeatability test showed that the RSD was 0.3%, indicating that the method had good repeatability. The stability test showed that the RSD was 0.4%, indicating that the test solution had good stability within 24h. The recovery rate was 97.2%and the RSD was 0.6%, indicating that the method was accurate and reliable. 6-gingerol's transfer rate ranged from 31.8%to 57.4%and the extraction rate was within the range of 9.6%-23.1%. The fingerprint similarity of 12 batches of Zingiberis Rhizoma standard decoction was>90%.Conclusion:The established quality control method for Zingiberis Rhizoma was stable and feasible; meanwhile, the standard preparation method for Zingiberis Rhizoma and its quality evaluation system were also established in this study.
Abstract：Objective:To obtain the glycosyltransferase gene involved in modification reaction of phytoalexin from Sorbus pohuashanensis suspension cell, and conduct sequence analysis and prokaryotic expression analysis.Method:Based on the transcriptome data, specific primers were designed to obtain 2 cDNA sequences of SaUGTs genes, construct prokaryotic expression vector HIS-MBP-pET28a-SaUGTs and induce the expression of recombinant SaUGTs protein.Result:SaUGT1 and SaUGT2 sequences were cloned and obtained from glycosyltransferases, then bioinformatic analysis of the sequence and prokaryotic expression analysis were conducted. SaUGT1gene contained 1 458bp open reading frame (ORF), encoding a polypeptide of 485 amino acids, with a relativemolecular weight of 54.27kDa and theoretical isoelectric point (pI) of 5.50.SaUGT2gene contained 1 431bp ORF, encoding a polypeptide of 476 amino acids, with a relativemolecular weight of 53.49kDa and theoretical pI of 5.63. Bioinformatics analysis indicated that SaUGT1 and SaUGT2 protein had no signal peptide, and the conserved domains of glycosyltransferase family were detected. Phylogenetic results showed that SaUGT1 and SaUGT2 proteins had the closest relationship with the UGT85 family of A. thaliana. Differential expression analysis revealed that the relative expression levels of SaUGT1 and SaUGT2 were increased significantly after being induced by yeast extract (YE), with the highest expression level found at 24h and 12h. The recombinant SaUGT1 and SaUGT2 proteins were successfully expressed in Escherichia coli DE3 cells and finally, the recombinant SaUGT1 and SaUGT2 proteins were purified through Ni2+ affinity chromatography.Conclusion:The glycosyltransferase gene was cloned from the S. aucuparia for the first time, and the prokaryotic expression vector was successfully constructed, laying foundation for further study of the function of this gene.
Abstract：Objective:To rapidly identify Cyathula officinalis and its adulterant C. capitata and C. officinalis*C. capitata and doped adulterant.Method:Properties combined with foam test method were used for identifying C. officinalis, its adulterant and doped adulterant. In the aspect of properties, 6 aspects including shape, size, texture, color, smell and taste, were observed, smelled and tasted. In the aspect of foam test, the volume of foam produced was used as the determination index to investigate the sample amount, water amount, shaking time, particle size, water temperature, repeatability, adulteration ratio and its stability.Result:In the aspect of properties, C. officinalis and its adulterant showed obvious difference in the shape, size, color, texture, smell and taste, especially the red color and bitter taste of its adulterant. In the aspect of foam test, the optimum parameters were as follows: sample particle (screened with 3sieves) 0.3g, a test tube with plug and scale, water 10mL and airtight, forced shaking up and down for 1min, settling for 5min. Such method can be used to identify C. officinalis, its adulterant and doped adulterant. The volume of foams produced by C. officinalis and its adulterant and different ratio of doped adulterant showed no change within 5-30min, slightly decreases after 9h; the higher adulteration ratio; the larger volume of foam and better stability. The 8 batches of C. officinalis and 8 batch of adulterants proved that the volume of the foams produced was all less than 2mL in the C. officinalis, more than 13mL in the adulterant is, and more than 5mL in 5% doped adulterant, showing statistical difference. From the properties combined with foam test, 5specific identification elements were obtained for identifying C. officinalis, its adulterant and doped adulterant.Conclusion:Through the 5specific identification elements, the properties combined with foam test can be used to distinguish the C. officinalis from its adulterant C. officinalis and C. officinalis*C. capitata and doped adulterant, characterized by accuracy, simpleness, short time, low cost and feasibility. It can provide a new method and reference for identifying C. officinalis from its adulterant and doped adulterant.
Abstract：Objective:To study the chemical constituents of Bletillae Formosanae Rhizoma and the distribution characteristics of active components in the fingerprint by establishing its high performance liquid chromatography (HPLC) fingerprints.Method:HPLC was used to establish the fingerprint of Bletillae Formosanae Rhizoma. Four reference substances, i.e. militarine, coelonin, 4-methoxy-9, 10-dihydrophenanthrene-1, 2, 7-triol and batatasin Ⅲ were used to identify chromatographic peaks. The fingerprints of 17 batches of Bletillae Formosanae Rhizoma fingerprints were analyzed and compared by "Computer-aided-similarity evaluation soft" and stoichiometry, and then compared with the fingerprint of Bletillae Rhizoma.Result:The established HPLC fingerprint method of Bletillae Formosanae Rhizoma showed good repeatability and stability. 20 common peaks were marked, four of which were identified by reference substances; militarine was No.8 common peak, and others corresponded to No. 10, No. 14 and No. 18 common peaks. Results showed that the similarities of samples except S4 were higher than 0.85, but the relative peak area of common peaks was quite different. Within the cluster distance 10, the samples are clustered into 5 categories, reflecting certain origin correlation. Principal component analysis (PCA) showed that the difference in samples was mainly caused by the common peaks located after No. 9 peak, where chemical constituents such as bibenzyl and dihydrophenanthrene were distributed. Bletillae Formosanae Rhizoma and Bletillae Rhizoma showed similar chemical constituents.Conclusion:The method provided a theoretical basis for the further clinical application and quality control of Bletillae Formosanae Rhizoma, as a substitute for Bletillae Rhizoma.
Abstract：Objective:To establish the HPLC fingerprint of wild and cultivated Notholirion bulbuliferum, and recognize them according to the chemical pattern, in the expectation of providing the basis for the quality control and domestication cultivation of N. bulbuliferum of origins.Method:Twenty samples of wild and cultivated N. bulbuliferum collected from different origins were detected by HPLC, and a common mode of fingerprint was established. The Similarity Evaluation System for Chromatographic Fingerprint of TCM (2012A edition) was used to evaluate the similarity of the samples. The differences among the samples were identified by chemical pattern recognition methods, including principal component analysis (PCA), cluster analysis (HCA) and partial least squares discriminate analysis (PLS-DA).Result:The HPLC fingerprint of N. bulbuliferum was obtained, and 26 common peaks were found in the chromatograph. Similarities of all samples were over 0.9, PCA, and HCA and PLS-DA results demonstrated obvious distinctions between wild and cultivated N. bulbuliferum. Eight constituents, such as pcoumaric acid were identified as biomarkers, representing major differences between the two varieties.Conclusion:The HPLC chromatogram of N. bulbuliferum developed in this paper has strong characteristics and repeatability. After being combined with the pattern recognition mode, it can be used as an effective method for evaluating the quality of N. bulbuliferum and distinguishing wild and cultivated N. bulbuliferum, and provide a reference for the quality control and domestication introduction of N. bulbuliferum.
Abstract：Objective:To study on the molecular mechanism of anti-platelet aggregation and anti-thrombosis of Sparganii Rhizoma.Method:Based on the data information and analysis functions of traditional Chinese medicine(TCM) database, TCM composition database and target database in TCM platform for integration of pharmacology, information of chemical compositions in Sparganii Rhizoma was retrieved, interaction network between potential targets of Sparganii Rhizoma and disease targets was built, and the key targets were enriched and calculated, the gene functions and pathways were analyzed, multidimensional relationship network of " herbal medicines-chemical components-key targets-key pathways" was built.Result:Active ingredients of Sparganii Rhizoma mainly included flavonoids and phenolic acids, such as mountain kaempferol, sanleng acid, linoleic acid, etc.Anti-thrombosis involved 202 key targets, including protein kinase Cδ(PRKCD), glucose kinase(GCK), (PRKAA2), adenosine ribonucleotide activated protein kinase α-2 catalytic subunit, etc; and it was related to the endocrine system, circulatory system, neurodegenerative diseases and other related biological processes and signal pathways.Anti-platelet aggregation involved 136 key targets, including PRKCD, cytochrome C oxidase protein 7C(COX7C), GCK, etc; and it was involved in Parkinson's disease, circulatory system, neurodegenerative diseases and other related biological processes and signal pathways.Conclusion:Sparganii Rhizoma regulates vascular endothelial cell adhesion molecule expression and platelet mitochondrial function mediated apoptosis of platelets mainly through regulating neurotransmitter activity in the central nervous system, in order to exert antithrombotic effect and anti-platelet aggregation effect.
Abstract：Ro Nus ZhurJes theory is the core content of Tibetan medicine, it can be divided into "taste performance" (Ro Yi Nus Pa) and "essential performance" (Ngo Bovi Nus Pa), and "taste performance" is divided into six tastes, eight properties, three tastes after digestion, seventeen effects and so on. "Essential performance" is divided into eight kinds, including odor efficacy, sources efficacy, color efficacy, et al.Form bearing relationships that "taste performance" and "essential performance" , the butt welding joins the three-factors and twenty characteristics of disease, it is the theoretical basis of Tibetan medicine from collection and processing, from formula to finished product. It is also the main method for the evaluation of drug performance by Tibetan medicine.Many classic Tibetan medicine prescriptions are compatible under the guidance of Ro Nus ZhurJes.However, at present, the research of Tibetan medicine mainly concentrates on the chemical composition and pharmacological mechanism.It is seldom studied from the Ro Nus ZhurJes theory of Tibetan medicine.It is difficult to find the core rules and effect of Tibetan medicine.Therefore, based on the ancient literature of Tibetan medicine, this paper combs the theory of Tibetan medicine property, explains the six tastes, three tastes after digestion, and seventeen effects of Tibetan medicine function in three-factors, twenty characteristics and other diseases attributes, in order to provide theoretical reference for the study of drug properties of Tibetan medicine.
Keywords：Tibetan medicine;The Four Medical Tantras;Ro Nus ZhurJes;taste performance;essential performance;six tastes;three tastes after digestion
Abstract：Objective:To explain the "multi-components, multi-targets, multi-pathways" mechanism of Erzhiwan in treating benign prostatic hyperplasia(BPH) based the network pharmacology.Method:Ingenuity pathway analysis(IPA) was used to construct components-targets-diseases network and PPI network, then the classified enrichment analysis of gene ontology (GO) and pathway enrichment analysis (KEGG) were carried out on the main function of its gene sets, so as to discuss the mechanism of Erzhiwan in the treatment of BPH.Result:Erzhiwan has 19 components in IPA; and apigenin, luteolin, oleanolic acid and quercitrin were common components of Ligustri Lucidi Fructus and Echiptae Herba and the main component of Erzhiwan. Muscarinic acetylcholine receptor M3 (CHRM3), muscarinic acetylcholine receptor M2 (CHRM2), urokinase plasminogen activator receptor (PLAUR), kinin releasing enzyme 3 (KLK3), cadherin 1 (CDH1), chemokines 3 (CCL3) and metalloproteinase-1 (MMP-1) were important targets for Erzhiwan to treat BPH. The target proteins in PPI network were enriched with 20 GO functions and 5 main KEGG pathways, and Docking was verified for relevant targets.Conclusion:Erzhiwan may play a role in treating BPH by activating MMP-1 and inhibiting KLK3 and CCL3 protein expressions, inducing apoptosis, inhibiting cell proliferation and intervening relevant pathways of mitogen-activated protein kinase/extracellular signal-regulated kinase(MAPK/ERK) and nuclear factor(NF)-κB(NF-κB).