Exploration of transcriptome SSR markers and its application in genetic diversity assessment of Asarum sieboldii / 中国中药杂志

To explore the genetic diversity of Asarum sieboldii this study developed SSR markers based on transcriptome sequencing results and five populations of A.sieboldii from different regions were used as samples for genetic diversity assessment using software such as GenALEx 6.5, NTSYS 2.1, and Structure 2.3.4. The results showed that 16 SSR markers with high polymorphism and good repeatability were selected from the A.sieboldii transcriptome. Primers designed based on the flanking sequences of these markers successfully amplified 56 polymorphic fragments from 150 individual samples of the five A.sieboldii populations. On average, each primer amplified 3.5 polymorphic fragments, ranging from 2 to 8. The mean values of expected heterozygosity(H_e), Shannon's diversity index(I), Nei's gene diversity index(H), and the polymorphic information content(PIC) were 0.172, 0.281, 0.429, and 0.382, respectively. The mean population differentiation coefficient(F_(ST)) was 0.588, consistent with the analysis of molecular variance(AMOVA) results, which indicated greater genetic variation among A.sieboldii populations(69%) than that within populations(31%). The percentage of polymorphic loci(PPL) ranged from highest to lowest as SNJ>LN>SY>SZ>TB. Principal coordinate analysis(PCoA) and UPGMA clustering analysis further revealed genetic clustering of A.sieboldii individuals based on their geographical distribution, consistent with the results of the structure clustering analysis. In summary, the SSR markers developed from the transcriptome effectively assessed the genetic differentiation and population structure of natural A.sieboldii populations, revealing a relatively low genetic diversity in A.sieboldii, with genetic variation primarily observed at the population level and a correlation between population differentiation and geographic distance.

Protective effect of Asarum polysaccharide on H1N1 influenza virus infection and expression of inflammatory factors / 中国中药杂志

In this paper, Asarum polysaccharides(AP) were extracted, and its composition was analyzed to study the activity against H1 N1 influenza virus in vitro and its intervention effect on mice with kidney Yang deficiency syndrome. AP was prepared by the strategy of water extraction and alcohol precipitation, the content was determined, and its monosaccharide composition was analyzed. The cell Real-time monitoring system and Reed-Muench model were adopted to evaluate the antiviral activity of AP in vitro. And the mouse model of kidney Yang deficiency syndrome was established in vivo to compare the efficacy of Mahuang Xixin Fuzi Decoction(MXF) and AP. MXF group and AP group were treated with clinical equivalent doses of 1.8 g·kg~(-1)·d~(-1) and 0.077 g·kg~(-1)·d~(-1) respectively, once a day for 6 consecutive days. Real-time PCR was used to detect the relative expression of M gene of H1 N1 influenza virus and cytokines in lung tissue. The content of AP in Asarum was 25.22%, and the protein content was 0.8%. And the monosaccharide composition was identified as L-rhamnose, D-arabinose, D-xylose, D-glucose, D-galactose and D-mannose. TI values of Tamiflu, MXF and AP were 30.00, 8.06 and 10.33, respectively. Three different doses of AP could significantly reduce the concentration of virus in supernatant. Compared with the model mice, lung indexes of MXF group and AP group decreased significantly(P<0.05), and the relative expression of M gene decreased significantly(P<0.05). The relative expressions of IL-10 and IFN-γ were up-regulated to varying degrees, while the relative gene expressions of IL-1β, IL-6 and MCP-1 were down-regulated to different degrees. In addition, AP could significantly enhance the expression of TNF-α(P<0.01). AP had a good anti-influenza virus activity in vitro, and could protect mice with kidney Yang deficiency syndrome by reducing the viral load in lung tissue, decreasing inflammation damage in lung tissue, and regulating the expression of inflammatory cytokines. Compared with the prescription of MXF, AP had a better antiviral activity.

Screening, identification and antagonistic effect evaluation of biocontrol strains to Asarum leaf blight / 中国中药杂志

Leaf blight is the main disease of Asarum. At present, chemical treatment is main measure for disease control, and there is no report on biological control. In order to achieve the biological control of Asarum leaf blight, the biocontrol strains with antagonistic effect on Asarum leaf blight were screened. The rhizosphere bacteria of healthy Asarum plants were isolated by soil dilution method, and the isolated strains were screened by the methods of antagonistic antifungal and fermentation liquid antifungal, then the strains were identified and the control effect in vivo was determined. Abiocontrol bacterial strains S2-31 which with high antagonism to leaf blight was obtained from more than 100 isolated strains. The inhibitory rates of antagonistic antifungal and fermentation liquid antifungal reached 92.47% and 60.56%, respectively. It was identified by morphology and 16 S rDNA sequence analysis, and the strain was identified as Brevibacillus laterosporus. The results of indoor potted experiment showed that the control effect was 79.87%, 71.44% and 66.82% on the 3 rd, 5 th and 7 th day after inoculation, respectively, which indicated that S2-31 could reduce the disease index and control the development of Asarum leaf blight.

Identification based on HPLC and anti-inflammatory targets as well as related constituents analysis of Asarum heterotropoides var. mandshuricum and A. sieboldii / 中国中药杂志

The present work is to establish an HPLC characteristic chromatograms of Asarum heterotropoides var. mandshuricum(AH) and A. sieboldii(AS), combined with cluster analysis for the identification of the two species, and predict their potential anti-inflammatory related targets by network pharmacological method. Eighty-nine samples(12 batches of AS and 77 batches of AH) were analyzed, and 11 characteristic peaks were identified by reference substances, UV spectrum and LC-MS. Cluster analysis showed that AS and AH were divided into two groups, and the ratio of characteristic peak areas can be used to distinguish them. When the ratio of characteristic peak sarisan to kakuol was greater than 5, it was AS, and when the ratio was less than 2, it was AH. The network pharmacological analysis of 119 constituents of Asari Radix et Rhizoma suggested that the anti-inflammatory effect of Asari Radix et Rhizoma might be related to COX-2, COX-1, iNOS, MAPK14, NR3 C1, PPARG and TNF. Among them, COX-2 is a relatively key target, which interacted with the characteristic constituents, asarinin, sesamin, safrole, methyleugenol and sarisan. The characteristic constituents asarinin and sesamin also interacted with the iNOS and MAPK14. Safrole and sarisan can also interact with iNOS, COX-1 and LAT4 H. Methyleugenol also showed interaction with COX-1 and LAT4 H. Since asarinin and sesamin interacted with three targets, COX-2, iNOS and MAPK14, it implied that they were the main active constituents for the anti-inflammatory activity of Asari Radix et Rhizoma. The COX-2 inhibitory activities of asarinin and sesamin were further studied by molecular docking and bioassay. The HPLC method established was simple, feasible and reliable, with predicted anti-inflammatory targets and anti-inflammatory constituents, which could provide a reference for improving the quality evaluation system of Asari Radix et Rhizoma.

Cloning of AhWRKY33 from Asarum heterotropoides and its genetic transformation in Arabidopsis / 中国中药杂志

The WRKY family genes, which play an important role in plant morphogenesis and stress response, were selected based on the data of the full-length transcriptome of Asarum heterotropoides. Using AtWRKY33, which regulates the synthesis of the camalexin in the model plant Arabidopsis to compare homologous genes in A. heterotropoides, primers were designed to amplify the open reading frame(ORF) fragment of AhWRKY33 gene by RT-PCR using total RNA of A. heterotropoides leaves as template. Real-time PCR results showed that there was a significant difference between the aerial part and the underground part of A. heterotropoides, the toxic aristolochic acid content is highly expressed in the leaves higher than the root. After verification, the WRKY33 gene of A. heterotropoides is ORF long 1 686 bp, encoding 561 amino acids.AhWRKY33 had two conserved WRKYGQK domains. According to the classical classification, it belongs to group Ⅰ WRKY transcription factor. A. heterotropoides WRKY33 had some homology with amino acids of other species. The study successfully constructed the plant eukaryotic expression vector PHG-AhWRKY33 and transformed Arabidopsis thaliana, the transgenic Arabidopsis was obtained by PCR detection and hygromycin resistant plate screening. It found that the germination of transgenic Arabidopsis seeds was accelerated and the stress resistance was increased. It laid a foundation for further analysis of WRKY transcription factor in the growth and development of A. heterotropoides and the synthesis of secondary metabolites.

Cytotoxic Constituents from the Roots of Asarum sieboldii in Human Breast Cancer Cells

Bioassay-guided fractionation of the roots of Asarum sieboldii led to the isolation of the six compounds methylkakuol (1), sesamin (2), asarinin (3), xanthoxylol (4), and (2E,4E,8Z,10E/Z)-N-(2-methylpropyl) dodeca-2,4,8,10-tetraenamide (5/6). Among the isolates, xanthoxylol (4) exhibited significant cytotoxicity against human breast cancer cells MCF-7 and MDA-MB-231 in vitro with IC₅₀ values of 9.15 and 13.95 µM, respectively.

Photosynthetic characteristics and active ingredients differences of Asarum heterotropoides var. mandshuricum under different light irradiance / 中国中药杂志

Chlorophyll content,leaf mass to per area,net photosynthetic rate and bioactive ingredients of Asarum heterotropoides var. mandshuricum,a skiophyte grown in four levels of solar irradiance were measured and analyzed in order to investigate the response of photosynthetic capability to light irradiance and other environmental factors. It suggested that the leaf mass to per area of plant was greatest value of four kinds of light irradiance and decreasing intensity of solar irradiance resulted in the decrease of leaf mass to per area at every phenological stage. At expanding leaf stage,the rate of Chla and Chlb was 3. 11 when A. heterotropoides var. mandshuricum grew in full light irradiance which is similar to the rate of heliophytes,however,the rate of Chla and Chlb was below to 3. 0 when they grew in shading environment. The content of Chla,Chlb and Chl( a+b) was the greatest value of four kinds of light irradiance and decreasing intensity of solar irradiance resulted in its decreasing remarkably( P<0. 05). The rate of Chla and Chlb decreased but the content of Chla,Chlb and Chl( a+b) increased gradually with continued shading. The maximum value of photosynthetically active radiation appeared at 10: 00-12: 00 am in a day. The maximum value of net photosynthetic rate appeared at 8: 30-9: 00 am and the minimum value appeared at 14: 00-14: 30 pm at each phenological stage if plants grew in full sunlight. However,when plants grew in shading,the maximum value of net photosynthetic rate appeared at about 10: 30 am and the minimum value appeared at 12: 20-12: 50 pm at each phenological stage. At expanding leaf stage and flowering stage,the average of net photosynthetic rate of leaves in full sunlight was remarkably higher than those in shading and it decreased greatly with decreasing of irradiance gradually( P < 0. 05). However,at fruiting stage,the average of net photosynthetic rate of leaves in full sunlight was lower than those in 50% and 28% full sunlight but higher than those in 12% full sunlight. All photosynthetic diurnal variation parameters of plants measured in four kinds of different irradiance at three stages were used in correlation analysis. The results suggested that no significant correlation was observed between net photosynthetic rate and photosynthetically active radiation,and significant negative correlation was observed between net photosynthetic rate and environmental temperature as well as vapor pressure deficit expect for 12% full sunlight. Positive correlation was observed between net photosynthestic rate and relative humidity expect for 12% full sunlight. Significant positive correlation was observed between net photosynthetic rate and stomatal conductance in the four light treatments. Only,in 12% full sunlight,the net photosynthetic rate was significantly related to photosynthetically active radiation rather than related to environmental temperature,vapor pressure deficit and relative humidity. In each light treatment,a significant positive correlation was observed between environmental temperature and vapor pressure deficit,relative humidity as well as stomatal conductance. Volatile oil content was 1. 46%,2. 16%,1. 56%,1. 30% respectively. ethanol extracts was 23. 44%,22. 45%,22. 18%,21. 12% respectively. Asarinin content was 0. 281%,0. 291%,0. 279% and 0. 252% respectively. The characteristic components of Asarum volatile oil of plant in different light treatments did not change significantly among different groups.

Research on inorganic elements of Asari Radix et Rhizoma from different localities and species / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the character of inorganic elements in Asari Radix et Rhizoma (Xixin).</p><p><b>METHOD</b>The contents of 53 inorganic elements in Xixin samples from different localities and species were determined by ICP-AES and ICP-MS. The statistical data were made using SAS.</p><p><b>RESULT</b>The result demonstrated that Xixin has the high contents of Fe, Cr, Li. It has been observed that the content of Cu and Pb of the samples are much higher than the standard level. The results of hierarchical cluster analysis revealed two groups which correspond with the species of the samples. No correlations between the contents of the inorganic elements and the localities of the samples were found. Some characteristic elements were displayed in some specific areas. The difference of the contents of the 53 inorganic elements between root and rhizome of Xixin was reported for the first time. The primary form of inorganic elements in Xixin has been studied for the first time. The result demonstrated that the extraction rate between different elements varied, with the average extraction rate of (22.25 +/- 24.96)%.</p><p><b>CONCLUSION</b>The inorganic elements analysis of Xixin can provide evidence of its identification, cultivation and application.</p>

Qualitative and quantitative analysis of dodecatetraenamides A, B in Asari Radix et Rhizoma / 中国中药杂志

To develop an analytic method for qualitative and quantitative analysis of dodecatetraenamides A, B in 42 samples of two official species of Asari Radix et Rhizoma( ARR) (37 samples of Asarum heterotropoides var. mandshuricum with different collection time and 5 samples of Asarum sieboldiivar. seoulense). The HPLC-IT-TOF-MS/MS methods for the qualitative and UPLC-PDA methods for the quantitative analysis were established. Dodecatetraenamides A, B were identified by comparing the retention time, UV absorption spectrum and quasi-molecular ion peak [ M + H]+ with the reference compound using HPLC-IT-TOF-MS/MS. The content of dodecatetraenamides A and B in ARR were determined by UPLC-PDA. The separation was successfully carried out on a ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 µm) column eluted with mobile phases of water (A) and acetonitrile (B) in gradient program (0-3 min, 35% B; 3-5 min, 35%-36% B; 5-6 min, 36%-43% B; 6 min-11 min 43% B; 11-12 min, 43%-100% B). The column temperature was 45 °C, and the detection wavelength was set at 254 nm. The flow rate was 0.6 mL · min(-1). On one level mass spectrometry scanning, the results showed that the quasi-molecular ion [M + H] + of both dodecatetraenamides A and B were m/z 248.20. The quantitative method with UPLC-PDA has made the baseline separation of the constituents, which were reported as mixtures in the most literatures. The average recovery of dodecatetraenamides A and B were 97.90% and 99.86%, the relative standard deviation were 0.4% and 1.1%, respectively. The contents of dodecatetraenamides A, B in all ARR samples was in the range of 0.11-3.89 and 0.24-6.65 mg · g(-1). Their contents reduced with the extension of storage time. Compared with the samples of 2013, the average content of the two constituents in the samples collected in year 2002-2003 reduced 34% and 36%, respectively (P < 0.05). Compared the A. sieboldii var. seoulense and A. heterotropoides var. mandshuricum with the same collective time and production area, the average contents of the two constituents in latter were up to (1.59 ± 0.75) mg · g(-1) and (2.90 ± 1.17) mg · g(-1), respectively, significantly higher than that in A. sieboldii var. seoulense (dodecatetraenamide A were (0.78 ± 0.52) mg · g(-1), dodecatetraenamide B were (1.69 ± 0.83) mg · g(-1)) (P < 0.05). The content of the dodecatetraenamide A in overground part was in the range of 0.11-0.33 mg · g(-1), dodecatetraenamide B was 0. 24-0.60 mg · g(-1), which were much lower than that of the underground part of ARR (dodecatetraenamide A was in the range of 0.73-3.89 mg · g(-1), dodecatetraenamide B was 2.11-6.24 mg · g(-1)). The method was certified to be simple, accurate and reliable and could be used for qualitative and quantitative analysis of dodecatetraenamide A and B in different species of ARR, also can be used for the comprehensive quality control of traditional Chinese medicine, Asari Radix et Rhizoma.

Development and validation of a HPLC-UV-ESI-MS method for the simultaneous quantitation of ten bioactive compounds in Dahuang Fuzi Tang / 中国天然药物

AIM@#To develop a high performance liquid chromatography (HPLC) coupled with electrospray ionization mass spectrometry (ESI-MS) and ultraviolet (UV) detector method for the acid-alkaline simultaneous determination of ten bioactive compounds, and analyze the effect of compatible medicinal plants on the concentration of components in Dahuang Fuzi Tang (DFT).@*METHOD@#The chromatographic separation was performed on a Hypersil BDS C18 analytical column by gradient elution with acetonitrile and formate buffer (containing 0.15% formic acid, V/V) at 25 °C with a flow rate of 1.0 mL·min(-1) and UV detection at 280 nm. Four of the ten compounds in DFT were identified and their MS fragments were elucidated by HPLC-ESI-MS, and the contents of the six compounds were determined by HPLC-UV.@*RESULTS@#All calibration curves showed good linear regression (r(2) ≥ 0.9990). The limits of detection and limits of quantification were 0.021-0.155 -g·mL(-1) and 0.076-0.520 -g·mL(-1), respectively. Overall precision RSD (intra-day and inter-day) were less than 2.96%, and the average recoveries were 98.35%-101.45%, with RSD ranging from 1.54% to 3.01% for the analytes.@*CONCLUSION@#The developed method can be applied for the quality control and provide analytical evidence on the chemical basis and combinational principles of DFT.

Investigation of anodized titanium implants coated with triterpenoids extracted from black cohosh: an animal study

PURPOSE: The purpose of this study was to evaluate bone response to anodized titanium implants coated with the extract of black cohosh, Asarum Sieboldii, and pharbitis semen. MATERIALS AND METHODS: Forty anodized titanium implants were prepared as follows: group 1 was for control; group 2 were implants soaked in a solution containing triterpenoids extracted from black cohosh for 24 hours; group 3 were implants soaked in a solution containing extracts of black cohosh and Asarum Sieboldii for 24 hours; group 4 were implants soaked in a solution containing extracts of pharbitis semen for 24 hours. The implants from these groups were randomly and surgically implanted into the tibiae of ten rabbits. After 1, 2, and 4 weeks of healing, the nondecalcified ground sections were subjected to histological observation, and the percentage of bone-to-implant contact (BIC%) was calculated. RESULTS: All groups exhibited good bone healing with the bone tissue in direct contact with the surface of the implant. Group 2 (52.44 +/- 10.98, 25.54 +/- 5.56) showed a significantly greater BIC% compared to that of group 3 (45.34 +/- 5.00, 22.24 +/- 2.20) with respect to the four consecutive threads and total length, respectively. The BIC% of group 1 (25.22 +/- 6.00) was significantly greater than that of group 3 (22.24 +/- 2.20) only for total length. CONCLUSION: This study did not show any remarkable effects of the extract of black coshosh and the other natural products on osseointegration of anodized titanium implants as coating agents. Further studies about the application method of the natural products on to the surface of implants are required.

Study on toxicity of uniform design-based compatible use of Veratrum nigrum and asarum / 中国中药杂志

The compatible use of Veratrum nigrum with asarum, is one of the eighteen incompatible pairs. To research the toxic regularity of the compatible use of V. nigrum and asarum, this experiment adopted the uniform design combining with acute toxicity test, with the number of died animals as the observation indicator. The results showed that the toxicity came from the common effect of V. nigrum and asarum, and V. nigrum after the compatible use, and V. nigrum made a greater contribution to the toxicity caused by the compatible use. But the toxicity did not absolutely enhance with the increase in use of V. nigrum.

Assessment on anti-nociception and anti-inflammation pharmacodynamics of Asarum heterotropoides var. mandshuricum and Asarum sieboldii / 中国中药杂志

<p><b>OBJECTIVE</b>To assess the anti-nociception and anti-inflammation pharmacodynamics of Asarum heterotropoides var. mandshuricum and A. sieboldii.</p><p><b>METHOD</b>Both the writhing test and hot plate test were conducted to assess the anti-nociceptive effect of Asarum and Xylene-induced mouse ear edema was conducted to assess the anti-inflammatory effect of Asarum.</p><p><b>RESULT</b>Twelve samples of A. heterotropoides var. mandshuricum and A. sieboldii from different producing areas showed anti-nociceptive and anti-inflammatory effects. Specifically, 27% to 61% of the seven samples of A. heterotropoides var. mandshuricum showed anti-nociceptive effect and while 34% to 48% of A. sieboldi showed anti-nociceptive effect. The inflammatory inhibition rate of A. heterotropoides var. mandshuricum produced in six producing areas (38%-57%) is higher than that of A. heterotropoides var. mandshuricum produced in five producing areas (34%-48%). The same kind of Asarum produced in different areas showed significant differences. A. heterotropoides var. mandshuricum produced in Jilin province (38%-57%) showed better anti-nociceptive effect than sample produced in Heilongjiang province (34%) in writhing test. A. heterotropoides var. mandshuricum produced in Heilongjiang (43%) province showed a better anti-nociceptive effect than samples produced in Liaoning province (29%-36%) in hot plate test. A. sieboldii produced in Shaanxi province (47%-49%) showed a better anti-nociceptive effect than samples produced in Hubei province (40%) in writhing test. A. sieboldii produced in Shaanxi province (45%-59%) showed better anti-nociceptive effect than samples produced in Chongqing (40%) in hot plate test. A. heterotropoides var. mandshuricum produced in Jilin province (51%-63%) showed better anti-inflammatory effect than samples produced in Heilongjiang province (50%). In totality, the results from analysis of geoherbalism showed that famous-region A. heterotropoides var. mandshuricum and A. sieboldii had a better anti-nociception effect than Asarum produced in other producing areas, famous-region A. heterotropoides var. mandshuricum had a better effect than those produced in other producing areas in anti-inflammation. But famous-region A. sieboldii showed no obvious difference from those produced in other producing areas in anti-inflammation.</p><p><b>CONCLUSION</b>All samples of Asarum showed anti-nociceptive and anti-inflammatory effects, but with significant differences among Asarum produced in different areas, indicating the eoherbalism to some extent.</p>

A new aristolochic acid derivative from Asarum himalaicum / 药学学报

To study the chemical constituents of Asarum himalaicum, fifteen compounds were isolated from a 70% ethanol extract by using a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, and semi-preparative HPLC. By spectroscopic techniques including 1H NMR, 13C NMR, and HR-ESI-MS, these compounds were identified as 4-demethoxyaristolochic acid BII (1), aristolochic acid I (2), aristolochic acid Ia (3), 7-hydroxyaristolochic acid I (4), aristolochic acid IV (5), aristolic acid II (6), debilic acid (7), aristololactam I (8), 9-hydroxyaristololactam I (9), 7-methoxyaristololactam IV (10), (2S)-narigenin-5, 7-di-O-beta-D-pyranosylglucoside (11), 4-hydroxybenzoic acid (12), 3, 4-dihydroxybenzoic acid (13), 4-hydroxycinnamic acid (14), and beta-sitosterol (15). All of these compounds (1-15) were obtained from A. himalaicum for the first time. Among them, 1 was identified as a new compound, and compounds 3-6, 9, 12-14 were isolated from Asarum genus for the first time. Since the kidney toxicity of aristolochic acids and aristololactams has been reported, the result of this investigation suggests that it should be cautioned to use A. himalaicum as a medicine.

Effect of light intensity on growth and quality of Asarum heterotropoides var. mandshuricum / 中国中药杂志

To study the infection rate of leaf spot disease, the drying rate of root and volatile oil content of Asarum heterotropoides var. mandshuricum at the unwrapping stage, blooming stage, the initial fruit stage, fructescence and wither stage during the growth period under the different sunlight intensity of 100% (I), 50% (II), 28% (III), 12% (IV). The volatile oil content was measured according to the method of Chinese Pharmacopoeia and the oil composition was determined by GC-MS. The unwrapping stage, blooming stage and the early fruit stage postponed about 2 days with decrease of the sunlight intensity. The infection rate of leaf was 88.46%, 70.00%, 0.23%, 0.07% under light intensity of I, II, III and IV, respectively, the drying rate was 25.14%, 28.27%, 30.23%, 31.57% under light intensity of I, II, III and IV, respectively, and the volatile oil content was 18.1, 17.6, 16.3, 15.3 mL x kg(-1) under light intensity of I, II, III and IV, respectively. The composition of the oil determined by GC-MS was different between the groups, but the content did not changed significantly with the decrease of the light intensity.

Anti-inflammatory and anti-nociceptive effects in mice of water and ethanol extracts of roots and rhizomes of Asarum heterotropoides var. mandshuricum / 中国中药杂志

<p><b>OBJECTIVE</b>To compare the anti-inflammatory and anti-nociceptive effects of the different water extracts which were prepared by regular decoction time or removing volatie oil and ethanol extracts which were prepared in different concentration ethanol of Xixin (the roots and rhizomes of Asarum heterotropoides var. mandshuricum), and then to investigate the anti-inflammatory and antinociceptive mechanisms.</p><p><b>METHOD</b>Xylene-induced ICR mouse ear edema and hot plate test were utilized to evaluate the anti-inflammatory and anti-nociceptive effects of Xixin at different dose level (water extracts of regular decoction time or removing volatie oil at the dose of 0.8 g x kg(-1) and 1.6 g x kg(-1); 95% ethanol extracts at the dose of 0.91 g x kg(-1) and 1.82 g x kg(-1); 50% ethanol extracts at the dose of 0.76 g x kg(-1) and 1.52 g x kg(-1)). Xylene-induced adrenalectomized mouse ear edema model was used to study the anti-inflammatory mechanisms. To reveal the anti-nociceptive mechanisms, mice were pretreated with naloxone in the hot plate test.</p><p><b>RESULT</b>The water extracts significantly reduced the weight of ear edema as compared with control group. Inhibition ratios were 43.20% and 63.69% at the higher dose, respectively. The ethanol extracts also significantly reduced the weight of ear edema and the inhibition ratios were 61.86% and 52. 56% at the higher dose, respectively. Mice administered water extracts delayed the latency times in the hot plate test. The anti-nociceptive effects of water extracts peaked at 2.0 h after i.g. administration. The hot plate latency times were increased by 51.27%, 62.78%, 60.08% and 68.00% at peak time, respectively. Regular decoction time group showed more significant effects in both models. The anti-inflammatory effect of 95% ethanol extracts was similar to 50% ethanol extracts. The water extracts were not effective in reducing xylene-induced adrenalectomized mouse ear edema. The anti-nociceptive effect of water extracts was blocked by naloxone.</p><p><b>CONCLUSION</b>Both the water extracts and ethanol extracts of Xixin showed considerable anti-inflammatory potency against xylene induced inflammation. The water extracts produced anti-nociception in thermal model. The water extracts prepared in regular decoction time showed better anti-inflammatory and anti-nociceptive effects. Both the 95% ethanol and 50% ethanol extracts showed similarly anti-inflammatory effects. The anti-inflammatory effect of water extracts related to adrenal gland. The anti-nociceptive effect of water extracts was involved in activating opioid receptor.</p>

Analysis of aristolochic acid A from the aerial and underground parts of Asarum by UPLC-UV / 药学学报

To develop an UPLC-UV method for the determination of aristolochic acid A in the aerial and underground parts of Asarum, an AcquityUPLC HSST3 column (50 mm x 2.1 mm ID, 1.8 microm) was used with the mobile phase of acetonitrile-0.1% FA and eluted in gradient mode. The flow rate was 0.5 mL min(-1), the detection wavelength was 251 nm, and the column temperature was 40 degrees C. The calibration curve was linear in the range of 18 - 1450 ng of aristolochic acid A with correlation coefficient of 0. 9999 and the minimum detective concentration was 18.25 ng mL (-1). The method developed is accurate and reproducible, and can be applied in the determination of aristolochic acid A in Asarum.

Effect of asarinin on the acute heart transplantation rejection and the expression of adhesion molecule / 中国中药杂志

<p><b>OBJECTIVE</b>To test the effect of asarinin, the extract of Herba Asari, on the acute heart transplantation rejection and the expression of adhesion molecule.</p><p><b>METHOD</b>Asarinin was extracted from herba asari. 64 SD rats undergone heart transplantation were divided into four groups: group A (control group), group B (Cyclosporine A treated), group C (Asarinin treated), and group D (1/2 CsA and 1/2 Asarinin). Some rats were used to examine survival time (n = 8) and the others were used to observe the pathological injury and the expression level of interrellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-I (VCAM-1) by using immunohistochemistry.</p><p><b>RESULT</b>Asarinin could prolong the survival time of allografts, which was similar to CsA group (P > 0.05). Asarinin could relieve the damage of cardiomyocytes of the transplanted. Asarinin could also decrease the level of ICAM-1 and VCAM-1 in the allografts.</p><p><b>CONCLUSION</b>Asarinin may play important roles in suppressing the immune rejection, prolong the allografts survival time and protect the donor organ, which was similar to CsA. The expression level of ICAM-1 and VCAM-1 is increased in suppressing the course of acute rejection and asarinin can inhibit their expression level. Asarinin can decrease the dosage of CsA.</p>

Studies on the botanical sources and DNA molecular identification of Herba Asari based on ITS sequence / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the genetic basis of using three species of Asarum as Herba Asari to determine the taxonomic positions of Asarum heterotropoides and A. siebodii; and to apply DNA molecular analysis as a tool for identification of Herba Asari.</p><p><b>METHOD</b>PCR, purification, sequence analysis were prerformed.</p><p><b>RESULT</b>MS sequences of the three Asarum species were obtained. 3 botanical sources of Herba Asari are closely clustered together on the topology tree; one inner branch is composed of A. heterotropoides and A. sieboldii, whereas another branch contains A. sieboldii. Their ITS sequences are different.</p><p><b>CONCLUSION</b>Three plant species of Herba Asari are closely related, and there are genetic reasons that they are used as the sources of the same medicine. The classification placement of A. sieboldii is not certain. The differences of ITS sequences of the botanical sources of Herba Asari can be used as a means of identification.</p>

Influence of Xinqin tablets on guinea-pig nasal hypersensitivity / 中国中药杂志

<p><b>OBJECTIVE</b>To study the influence of Xinqin tablets on guinea-pig nasal hypersensitivity.</p><p><b>METHOD</b>2,4-Toluene Diisocyanate (TDI) was selected as antigen and used in nose to establish guinea-pig allergic rhinitis. The effects of Xinqin tablets on symptoms of nasal hypersensitivity in guinea-pigs, histamine content of nasal mucosa and activity of nitric oxide synthase (NOS) were examined.</p><p><b>RESULT</b>Xinqin tablets could significantly relieve the pathological symptoms of nasal hypersensitivity in guinea-pig, reduce histamine content of nasal mucosa and inhibit the activity of nitric oxide synthase.</p><p><b>CONCLUSION</b>Xinqin tablets have significant effect on nasal hypersensitivity, and prevent the occurrence of allergic rhinitis.</p>