Transcriptomic analysis in Anemone flaccida rhizomes reveals ancillary pathway for triterpene saponins biosynthesis and differential responsiveness to phytohormones / 中国天然药物

Anemone flaccida Fr. Schmidt is a perennial medicinal herb that contains pentacyclic triterpenoid saponins as the major bioactive constituents. In China, the rhizomes are used as treatments for a variety of ailments including arthritis. However, yields of the saponins are low, and little is known about the plant's genetic background or phytohormonal responsiveness. Using one-quarter of the 454 pyrosequencing information from the Roche GS FLX Titanium platform, we performed a transcriptomic analysis to identify 157 genes putatively encoding 26 enzymes involved in the synthesis of the bioactive compounds. It was revealed that there are two biosynthetic pathways of triterpene saponins in A. flaccida. One pathway depends on β-amyrin synthase and is similar to that found in other plants. The second, subsidiary ("backburner") pathway is catalyzed by camelliol C synthase and yields β-amyrin as minor byproduct. Both pathways used cytochrome P450-dependent monooxygenases (CYPs) and family 1 uridine diphosphate glycosyltransferases (UGTs) to modify the triterpenoid backbone. The expression of CYPs and UGTs were quite different in roots treated with the phytohormones methyl jasmonate, salicylic acid and indole-3-acetic acid. This study provides the first large-scale transcriptional dataset for the biosynthetic pathways of triterpene saponins and their phytohormonal responsiveness in the genus Anemone.

Reversible ON-OFF switching of single-molecule-magnetism associated with single-crystal-to-single-crystal structural transformation of a decanuclear dysprosium phosphonate.

{Dy5(EDDC)2(µ3-AcO)2(µ5-C15H11PO3)(µ4-C15H11PO3)(µ2-AcO)2(AcO)2(H2O)(CH3OH)2}2(µ4-C2O4)·xH2O (I), where H2EDDC is N',N'',E,N',N'',E-N',N''-(ethane-1,2-diylidene)dipyrazine-2-carbohydrazide and C15H11PO3H2 is 9-anthrylmethylphosphonic acid, is found to undergo two consecutive single-crystal-to-single-crystal transformations. The first is under UV irradiation (λ = 365 nm for 3 d in air) to {Dy5(EDDC)2(µ3-AcO)2(µ5-C15H11PO3)2(µ2-AcO)2(AcO)2(H2O)3}2(µ4-C2O4)·xH2O (I-UV) where the two CH3OH are replaced by two H2O and the second by annealing under N2 at 100 °C on a diffractometer or under Ar in a glovebox to {Dy5(EDDC)2(µ3-AcO)2(µ5-C15H11PO3)2(µ2-AcO)4(H2O)}2(µ4-C2O4) (I-A-N2 or I-A-Ar) where it has lost two H2O molecules. The second transformation is reversible by exposure to air at room temperature (I-A-N2-cool). While the overall structures are the same (retaining the space group P21/c), there is a considerable expansion of the unit cell from I (8171 Å3) to I-UV (8609 Å3) and I-A-N2 (8610 Å3) and the coordinations of the Dy atoms undergo major reconstructions. This is associated with switching the single-molecule-magnetism (SMM) from OFF for I to ON for I-UV and to OFF again for I-A-Ar in air. Such a switching mechanism associated with the retention of crystallinity is unique in the chemistry of dysprosium. The structure of the molecule is formed from two symmetry related pentamers joined by an oxalate. A related compound containing two isolated neutral pentamers {Dy5(EDDC)2(µ3-AcO)2(µ5-C15H11PO3)2(µ2-AcO)3(AcO)2(H2O)2}{Dy5(EDDC)2(µ3-AcO)2(µ5-C15H11PO3)(µ4-C15H11PO3)(µ2-AcO)3(AcO)2(CH3OH)2}·2CHCl3 (II) has also been isolated with closely related Dy coordination and it exhibits similar SMM behaviour in zero field.

Single-Molecule Magnet Behavior of 1D Coordination Polymers Based on DyZn2(salen)2 Units and Pyridin- N-Oxide-4-Carboxylate: Structural Divergence and Magnetic Regulation.

Two 1D coordination polymers composed of DyZn2(salen)2 units and pyridin- N-oxide-4-carboxylate have been prepared by solvothermal reactions. Complex 1 with the formula {[DyZn2(La)2(POC)](OH)(ClO4)}·H2O·MeOH [H2La] = N, N'-bis(3-methoxysalicylidene)-1,3-diaminopropane, POC- = pyridin- N-oxide-4-carboxylate] is composed of a zigzag chain cation [DyZn2(La)2(POC)] n2 n+ as well as isolated hydroxide and perchlorate anions. Complex 2 with the formula {[Dy3Zn7(Lb)6(POC)6](OH)3(ClO4)2}·9H2O [H2Lb] = N, N'-bis(3-methoxysalicylidene)-1,2-diaminoethane] is also an ionic compound containing isolated hydroxide and perchlorate anions, but its cation shows a novel nanowire structure, in which six-coordinate zinc(II) ions with C3 symmetry act as the axis and [DyZn2(Lb)2(POC)]2+ structural units as spiral leaves. Complex 1 shows good single-molecule magnet performance with an Ueff/ k value of 235.3(3.1) K ( Hdc = 0 Oe), one of the largest values for coordination polymers. A butterfly-shaped magnetic hysteresis loop can be monitored at as high as 3.8 K for 1, while a dc field is necessary for complex 2 to display slow magnet relaxation owing to the quantum-tunneling effect, with a much smaller Ueff/ k value of 14.6(0.2) K ( Hdc = 1000 Oe). The difference of magnetic properties has been explained using detailed ab initio calculations.

Bone marrow mesenchymal stem cell transplantation inhibits apoptosis in the rat spinal cord injured by acrylamide / 中国组织工程研究

BACKGROUND: Until now, there is no effective treatment for peripheral neuropathy caused by acrylamide. Therefore, it is necessary to explore new treatment methods. OBJECTIVE: To explore the protection role and its mechanism of bone marrow mesenchymal stem cells (BMSCs) against acrylamide-induced intoxication in the spinal cords of rats. METHODS: BMSCs were cultured by the whole bone marrow adherence method and identified by morphological observation and flow cytometry detection. Thirty Sprague-Dawley rats, clean grade, were randomly divided into three groups (n=10 for each group): normal control group, acrylamide group and BMSCs transplantation group. The latter two groups received acrylamide by gavage, 50 mg/(kg?d), 5 days per week, for 2 weeks with an interval of 2 days. Then, in the BMSCs transplantation group, 3×106BMSCs were transplanted by the caudal vein, 5 days per week, for 3 consecutive weeks. Hematoxylin-eosin staining was utilized to observe the morphological changes of the spinal cord. Tunel assay was used to detect cell apoptosis. Western blot assay was adopted to detect the expression levels of Bcl-2 and Bax. RESULTS AND CONCLUSION: In the acrylamide-exposed rats, the damage to the structure was found in the spinal cords by morphological observation, which was significantly alleviated after BMSCs transplantation. The disturbed expression levels of Bax and Bcl-2 were also significantly inversed after BMSCs transplantation (P < 0.05). These results suggest that BMSCs transplantation can inhibit cell apoptosis in the spinal cords of acrylamide-intoxicated rats, probably by up-regulating expression of Bcl-2 and down-regulating expression of Bax.

Side chain carbon losssalidroside analogues (N04) protects EAhy926 human endothelial cells against hypoxia injury by regulating the expressions of HIF-1α-related factors / 医学研究生学报

Objective Salidroside is a major active component of integripetal rhodiola herbal medicine, which has a significant activity against hypoxia and ischemia.This study was to investigate the effects of side chain carbon losssalidroside analogues (N04) on the expressions of HIF-1α-related factors in the hypoxia-injured EAhy926 human vascular endothelial cells.Methods EAhy926 human umbilical vein endothelial cells in the logarithmic growth phase were randomly divided into a normal control, a hypoxia model control, a salidroside, a high-dose N04, a medium-dose N04, and a low-dose N04 group.The hypoxia model was established by depriving the culture medium of sugar and serum and culturing the EAhy926 cells in an environment of 95%N2+5%CO2 for 2 hours, followed by intervention with salidroside at 1×10-6 mol/L and N04 at 1×10-6, 1×10-7, and 1×10-8 mol/L, respectively.Then, the activity of the cells was detected by MTT assay, their LDH activity examined by spectrophotometry, the mRNA expressions of HIF-1α and VEGF measured by RT-PCR, the protein expressions of HIF-1α, VEGF and pVHL determined by Western blot, and the activity of eNOS measured by ELISA.Results Compared with the normal control group, the hypoxia model cells showed significantly reduced activity (0.51±0.05 vs 0.27±0.02, P<0.01), an elevated LDH level ([6.65±1.43] vs [78.82±2.33] U/L, P<0.01), and decreased eNOS activity ([1.56±0.23] vs [1.16±0.20] U/100 mL, P<0.01).In comparison with the hypoxia model group, the cells treated with high-, medium-, and low-dose N04 exhibited remarkably increased activity (0.27±0.02 vs 0.0.42±0.05, 0.40±0.03 and 0.37±0.04, P<0.01), a reduced LDH level ([78.82±2.33] vs [53.05±3.90], [58.42±4.45] and [62.73±3.63] U/L, P<0.01), and increased eNOS activity ([1.16±0.20] vs [3.01±0.47], [2.60±0.26] and [2.32±0.29] U/100 mL, P<0.01).The activity of eNOS was also increased in the salidroside group ([2.32±0.29] U/100 mL, P<0.01).The cell activity in the high-and medium-dose N04 groups was markedly higher than that in the salidroside group (P<0.05), and so was the eNOS activity in the high-dose N04 group and the LDH level in the medium-and low-dose N04 groups (P<0.05).In comparison with the normal control group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were significantly up-regulated in the hypoxia model group (P<0.01) while that of the pVHL protein markedly down-regulated (P<0.01).Compared with the hypoxia model group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were remarkably reduced (P<0.05), while that of the pVHL protein markedly elevated (P<0.05).Both the expressions of VEGF mRNA and HIF-1α protein were significantly lower in the medium-and low-dose N04 groups than in the salidroside group (P<0.05).Conclusion N04 can protect vascular endothelial cells against hypoxia-induced injury by regulating the expression of HIF-1α-related factors and eNOS.

Medicine packaging improvement based on PAP shelter hospital requirements / 医疗卫生装备

Objecive To explore the medicine packaging improvement based on PAP shelter hospital requirements to fulfill emergency medicine support,Methods The medicine support during rescue was discussed from the aspects of considerations in medicine plan,requirement for emergency medicine,special medicine purchasing and medicine support,and some countermeasures were put forward for improving medicine packaging.Results Its suggested that the optimization of medicine packaging be performed with considerations on medicine property,package capacity,convenience,transport and environmental suitability.Conclusion Emergency medicine support is of importance for disaster medical rescue,and medicine packaging improvement based on PAP shelter hospital requirements contributes to enhancing the efficiency during disaster rescue.

An experimental study on inhibitory effects and mechanisms of N-butanol extract of Potentilla anserine L. against hypoxia-induced nitric oxide on hippocampus neurons of rats / 中国中西医结合急救杂志

Objective To study in vitro the inhibitory effects and mechanisms of N-butanol extract of Potentilla anserine L.(NP)against hypoxia-induced nitric oxide(NO)in hippocampus neuron of rats. Methods The models of hippocampus neurons hypoxia injury of Sprague-Dawley(SD)neonatal rats were cultured in vitro. The cultured hippocampus neurons were divided randomly into blank control group, hypoxia injury model group, nimodipine group(2 μmol/L)and NP high(250.0 mg/L),middle(62.5 mg/L),low(15.6 mg/L)dose groups. The activities of hippocampus neurons were examined by methyl thiazolyl tetrazolium(MTT)assay,and meanwhile their contents of nitrogen monoxidum(NO)were detected. Half quantity reverse transcription-polymerase chain reaction(RT-PCR)and Western blotting were used to detect neuronal nitric oxide synthetase(nNOS)mRNA and protein expression levels respectively in each group,immunocytochemistry stain was used to detect protein positive rate. Results Compared with blank control group,the activity of neuron〔absorbance(A)value〕was significantly decreased(0.0826±0.0095 vs. 0.3315±0.0105),content of NO(μmol/g:0.0509±0.0027 vs. 0.0291±0.0032), the expression levels of nNOS mRNA (0.1463±0.0081 vs. 0.0801±0.0058), the positive rate of nNOS〔(74.4238±3.9423)%vs.(28.3714±4.1361)%〕,the expression levels of nNOS protein(A value:1.9130±0.0471 vs. 0.5068±0.0368)were all significantly increased in the hypoxia injury model group(all P0.05). Conclusions NP can ameliorate the injury of rat hippocampus neurons induced by hypoxia in vitro. The possible mechanisms might be related to the effective inhibition of the synthesis of nNOS and NO excessive generation.

Oxido[N-(2-oxidobenzyl-idene-κO)leucinato-κ²N,O](1,10-phenanthroline-κ²N,N')vanadium(IV).

In the title V(IV) complex, [VO(C13H15NO3)(C12H8N2)], the oxidovanadium cation is N,N'-chelated by a 1-10-phenanthroline ligand and N,O,O'-chelated by a Schiff base anion in a distorted octa-hedral geometry. Weak inter-molecular C-H⋯O hydrogen bonds occur in the crystal structure which contains solvent-accessible voids of 81 ų.

Genetic diversity of Atractylodes macrocephala by ISSR / 中国中药杂志

<p><b>OBJECTIVE</b>To assess the genetic diversity of Atractylodes macrocephala in 12 cultivated populations and 3 wild populations.</p><p><b>METHOD</b>The molecular marker technique ISSR was used to investigate the genetic diversity of 365 samples from 15 populations in Zhejiang, Anhui and Hebei Provinces. Data was analyzed by POPGEN 32, and a cluster diagram was presented by UPGMA.</p><p><b>RESULT</b>One hundred and two amplified fragments were obtained using 13 ISSR primers. 94 polymorphic loci were identified as 92.16%. Nei's genetic diversity index (He) was 0.406 5, Shannon diversity index (I) was 0.590 3, and the coefficient of gene differentiation (Gst) was 0.202 5. The genetic similarity coefficient among the populations ranged from 0.690 7 to 0.960 5 in an average of 0. 825 6. No significant difference was found among the populations based on the clustering analysis.</p><p><b>CONCLUSION</b>There is a low level of genetic differentiation in higher genetic diversity of the A. macrocephala populations. No obvious regional or species diversities were identified. This is consistent with current status of cultivation and gerplasm circulation in A. macrocephala.</p>

3-Phenyl-2-(pyrrolidin-1-yl)-5,6-dihydro-8H-thio-pyrano[4',3':4,5]thieno[2,3-d]pyrimidin-4(3H)-one.

In the title compound, C(19)H(19)N(3)OS(2), the thio-pyran ring adopts a twist-chair conformation and the pyrimidinone unit is essentially planar, with a mean deviation of 0.0497 Å. The thio-phene ring is essentially planar with a maximum deviation of 0.024 (2) Å, while the pyrrolidine ring exhibits an envelope conformation. The pyrimidinone and thio-phene rings are almost coplanar, forming a dihedral angle of 6.31 (15)°, while the dihedral angle between the mean planes of the phenyl ring and the pyrimidinone ring is 68.13 (10)°. In the crystal structure, adjacent mol-ecules are linked by C-H⋯O hydrogen bonds, forming a two-dimensional network in the ac plane.

(Amino-acetato-κO,N)bis-(quinolin-8-olato-κO,N)cobalt(III) methanol solvate.

In the crystal structure of the title compound, [Co(C(2)H(4)NO(2))(C(9)H(6)NO)(2)]·CH(3)OH, the Co(III) atom is chelated by two quinolin-8-olate and one glycinate anions in a distorted octa-hedral coordination geometry. The five-membered chelating glycinate ring assumes an envelope conformation. The complex mol-ecules are assembled by inter-molecular N-H⋯O hydrogen bonding.