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Artemia franciscana, native to America, has recently colonized as non-indigenous population in Asia, Europe, North Africa, and Australia. We evaluated the effects of the colonization of A. franciscana on genetic differentiation in new environments in the United Arab Emirates (UAE). We used the COI marker to determine the genetic structure and origins of exotic populations in the UAE. Results confirmed the colonization of A. franciscana in two localities. Invasive populations of A. franciscana had significantly lower genetic variation than native populations in the Great Salt Lake and San Francisco Bay. Results showed that the studied populations could not have colonized directly from natural American habitats, and they possibly were from secondary introduction events of other non-indigenous populations. Genetic analysis yielded different demographic patterns for the studied invasive populations. The population in Al Wathba Wetland Reserve (AWWR) demonstrated demographic expansion, whereas in Godolphin Lakes (GL), it reached a demographic equilibrium. Neutrality tests showed an excess of recent and historical mutations in the COI gene pool of invasive AWWR Artemia in the new environment. The results suggest that different ecological conditions in new environments can exert selective pressures during the introduction of an exotic population, which can affect genetic variation.

To facilitate sustainable agriculture in the acidic soil in the southern region of China, reducing inorganic fertiliser in combination with an alkaline humic acid fertiliser amendment (HAFA) was in...

Artemia is the most common live food that is used in aquaculture worldwide. This study reports on biometrical variation in the cysts of the introduced, originally American Artemia franciscana from 24 non-native localities and two native habitats, in Asia and the U.S.A., respectively. The results show, that the largest diameter of untreated cysts, the largest diameter of decapsulated cysts, and a thicker chorion usually are found in invasive populations. Because of the small cysts, which have an effect on an increasing quantity per unit weight and thus could be the cause of increased hatching efficiency, commercial productions of A. franciscana cysts from native sources should potentially be considered higher quality than productions from non-indigenous environments. Principal Component Analysis revealed that all cyst batches from San Francisco Bay were classified in one group and most of the invasive populations could be arranged in another, separate group. Although the diameter of the decapsulated cyst and chorion thickness showed a negative and significant correlation among invasive populations, there was no significant relationship within native populations. These observations contrast with biometrical patterns of parthenogenetic populations.

The complete mitochondrial genome of Sinularia penghuensis was sequenced and analyzed using next-generation sequencing. The present mitochondrial genome was 18730 bp in length, containing 14 protein-coding genes (PCGs) (cox1-cox3.nad1-nad6, nad4L, atp6, atp8, cytb, and MutS), two ribosomal RNA genes (rRNAs) (12S and 16S), and one transfer RNA gene (Met-tRNA). The phylogenetic analysis of family Alcyoniidae revealed that S. penghuensis and Sinularia maxima cluster together. Five species in Sinularia reveals high identity in mitogenome sequences that the lowest variable sites (SNPs) were found between S. penghuensis and S. maxima.

The transdermal route of administration provides numerous advantages over conventional routes i.e., oral or injectable for the treatment of different diseases and cosmetics applications. The skin also works as a reservoir, thus deliver the penetrated drug for more extended periods in a sustained manner. It reduces toxicity and local irritation due to multiple sites for absorption and owes the option of avoiding systemic side effects. However, the transdermal route of delivery for many drugs is limited since very few drugs can be delivered at a viable rate using this route. The stratum corneum of skin works as an effective barrier, limiting most drugs’ penetration posing difficulty to cross through the skin. Fortunately, some non-invasive methods can significantly enhance the penetration of drugs through this barrier. The use of nanocarriers for increasing the range of available drugs for the transdermal delivery has emerged as a valuable and exciting alternative. Both the lipophilic and hydrophilic drugs can be delivered via a range of nanocarriers through the stratum corneum with the possibility of having local or systemic effects to treat various diseases. In this review, the skin structure and major obstacle for transdermal drug delivery, different nanocarriers used for transdermal delivery, i.e., nanoparticles, ethosomes, dendrimers, liposomes, etc., have been discussed. Some recent examples of the combination of nanocarrier and physical methods, including iontophoresis, ultrasound, laser, and microneedles, have also been discussed for improving the therapeutic efficacy of transdermal drugs. Limitations and future perspectives of nanocarriers for transdermal drug delivery have been summarized at the end of this manuscript.

In order to study the Sr content and hydrochemical characteristics of shallow groundwater in Tongren area of Guizhou province, we investigated 168 groups of shallow groundwater. We take the wall rocks are divided into sedimentary environment units. The difference environmental units distribution and evolution of Sr2+ in groundwater were studied by means of mathematical statistics, graphical analysis, hydrochemical analysis and ion ratio. The conclusion is as follows : (1) the main source of Sr2+ in groundwater in the study area is rock mineral weathering dissolution or dissolution, and its concentration is significantly affected by rock composition, which occurs in the platform strata and is controlled by carbonate rock weathering, and the clastic rock sedimentary environment is significantly affected by silicate weathering. (2) The concentration of strontium varies greatly in each environmental unit, among which the concentration is the highest and relatively uniform in the semi-confined platform environment, with a range of 0.196~0.362 mg/L, and the background range is only 0.015~0.059 mg/L in the restricted platform environment. However, due to the presence of a large number of rock salt mineralization points, the local maximum value is abnormal.

To investigate whether miRNA-30a is involved in the pathogenesis of ITP by affecting the differentiation of Th17 cells, and to explore its possible mechanism of miRNA-30a involved in the pathogenesis of ITP through the verification of the target gene SOCS3 for the prediction of miRNA-30a.Firstly, a chronic ITP mouse model was established. The expression of miRNA-30a and RORγt in the spleen mononuclear cells were detected and their correlation were analyzed. Secondly, the luciferase vector containing 3'UTR of the target gene and green fluorescent vector containing miRNA were constructed. Luciferase fluorescence detection, real-time fluorescent quantitative PCR (qPCR) and Western blot were used to verify whether SOCS3 is the target gene of miRNA-30a.The platelet count of mice in experimental group decreased to below 20% of normal ones after 48 hours of injection of anti-mouse platelet serum (APS), which was maintained for 14 days at least; the expression of miRNA-30a and RORγt in the spleen mononuclear cells in experimental group were higher than those in the control group(P＜0.05), moreover, there was a positive correlation between them (r=0.54); the activity of luciferase in PMDH-GFP-miRNA-30a and pMIR-report-UTR was significantly lower than that in PMDH-GFP empty plasmid and pMIR-report-UTR(P＜0.05); The expression of SOCS3 at mRNA and protein level was not different from that in the control group.Chronic ITP mouse model has been established successfully; miRNA-30a expression in spleen mononuclear cells of ITP mouse increase, and positively correlated with the expression of RORγt, which contribute to the pathogenesis of ITP by affecting the differentiation of Th17 cells; SOCS3 is able to bind to the target site of miRNA-30a, but might not be its functional target gene.miRNA-30a通过介导Th17细胞分化影响免疫性血小板减少性紫癜发病的初步探讨.探讨miRNA-30a是否通过影响Th17细胞分化参与ITP发病；通过对miRNA-30a预测靶基因SOCS3的验证，进一步探讨miRNA-30a参与ITP发病的可能机制.建立慢性ITP小鼠模型，检测其脾脏单个核细胞中miRNA-30a、 RoRγt的表达并分析两者相关性；构建含靶基因mRNA 3′UTR的荧光素酶表达载体及含有miRNA的绿色荧光载体，通过Luciferase荧光检测、实时荧光定量PCR（qPCR）、Western blot验证SOCS3是否为miRNA-30a的靶基因.实验组小鼠血小板计数在注射抗小鼠血小板血清（APS）48 h后下降至正常的20%，且至少可维持14 d。实验组小鼠脾脏单个核细胞中miRNA-30a及RoRγt表达较对照组升高（P＜0.05），且两者存在正性相关（r=0.54）。pMDH-GFP-miRNA-30a与pMIR-report-UTR实验组的荧光素酶的活性明显低于pMDH-GFP空质粒与pMIR-report-UTR的对照组（P＜0.05）。转染miRNA-30a质粒的EL4细胞中SOCS3在mRNA及蛋白水平与对照组均无差异.miRNA-30a在ITP小鼠脾脏单个核细胞中表达升高，且与RORγt表达呈正相关性，它可能通过影响Th17细胞分化参与ITP发病；SOCS3能与miRNA-30a靶位点结合，却不是其功能靶基因.

BACKGROUND: Immune imbalance of regulatory T cells (Treg)/T helper 17 cells (Th17) contributes to the development of immune thrombocytopenic purpura (ITP). The dysregulation of miRNAs is important in the development of ITP. However, the role of miR-106b-5p in Treg/Th17 imbalance remains unknown in ITP. MATERIALS AND METHODS: Peripheral blood was collected from patients with ITP and healthy controls, and CD4 + T cells were further isolated. miR-106b-5p, nuclear receptor subfamily 4 group A member 3 (NR4A3), forkhead box protein 3 (Foxp3), IL-17A, and TGF-β expressions were detected by qRT-PCR, western blot, or ELISA. The effect of miR-106b-5p on NR4A3 was detected by dual-luciferase reporter gene assay. RESULTS: Compared with healthy controls, miR-106b-5p was elevated in peripheral blood of patients with ITP, and NR4A3 expression was decreased. sh-NR4A3 significantly decreased Foxp3 and TGF-β expressions, indicating that NR4A3 may regulate Treg differentiation via Foxp3. Additionally, NR4A3 was identified to be a target of miR-106b-5p, and miR-106b-5p was able to negatively modulate NR4A3 expression. Moreover, we found miR-106b-5p induced immune imbalance of Treg/Th17 through NR4A3. In vivo experiments revealed that silencing miR-106b-5p promoted Treg differentiation and increased the number of platelets, suggesting the relief of ITP. CONCLUSION: miR-106b-5p regulated immune imbalance of Treg/Th17 in ITP through the NR4A3/Foxp3 pathway.

OBJECTIVE: To investigate the protective effects of paeoniflorin against PM2.5-induced damage in BEAS-2B cells and explore the possible mechanism. METHODS: With a factorial design, this study was performed to observe the protective effects of different doses of paeoniflorin against PM2.5-induced BEAS-2B cell growth inhibition and the effects of paeoniflorin on the contents of malondialdehyde (MDA) and intracellular reactive oxygen species (ROS) in the cell cultures. RESULTS: Exposure to increased PM2.5 concentrations caused significant decrease in the cell survival rate (P < 0.05) with a clear dose-response relationship (r=-0.759, P < 0.05). Treatment of the cells with paeoniflorin significantly attenuated PM2.5-induced inhibition of BEAS-2B cell survival (P < 0.05), but the effect of paeoniflorin was not dose-dependent (P > 0.05). PM2.5 exposure also significantly increased the contents of MDA and intracellular ROS (P < 0.05), and paeoniflorin obviously antagonized these effects of PM2.5. CONCLUSION: Paeoniflorin can protect BEAS-2B cells from PM2.5-induced growth inhibition, and the mechanism might be related to the anti-oxidant effects of paeoniflorin.

A bioactive chromone, 8-C-glucosyl-7-O-methyl-(S)-aloesol (compound 1), was prepared by the acid-catalyzed hydrolysis of isoaloeresin D in Aloe barbadensis Miller. The hydrolysis conditions were successfully optimized by response surface methodology (RSM). The results indicated that RSM in the present study was adequate and effective. Then, reversed-phase flash chromatography was utilized for the isolation. In a one-step operation within 45 min, 26 mg of compound 1 was isolated from 153 mg of hydrolysis product, with a purity of 99.3%. These achievements would afford sufficient amount of high pure compound for chromatographic purpose and pharmacological activity screening.

Methamphetamine (METH) belongs to Amphetamine-type stimulants, METH abusers are at high risk of neurodegenerative disorders, including Parkinson's disease (PD). However, there are still no effective treatments to METH-induced neurodegeneration because its mechanism remains unknown. In order to investigate METH’s neurotoxic mechanism, we established an in vitro PD pathology model by exposing PC12 cells to METH. We found the expression of nitric oxide synthase (NOS), nitric oxide (NO) and α-synuclein (α-syn) was significantly increased after METH treatment for 24 h, in addition, the aggregattion of α-syn and the S-nitrosylation of protein disulphideisomerase(PDI) were also obviously enhanced. When we exposed PC12 cells to the NOS inhibitor N-nitro-L-arginine(L-NNA) with METH together, the L-NNA obviously inhibited these changes induced by METH. While when we exposed PC12 cells to the precursor of NO L-Arginine together with METH, the L-Arginine resulted in the opposite effect compared to L-NNA. And when we knocked down the PDI gene, the L-NNA did not have this effect. Therefore, PDI plays a significant role in neurological disorders related to α-syn aggregation, and it suggests that PDI could be as a potential target to prevent METH-induced neurodegeneration.

ObjectiveTo study the effect of Qilongtoutong granule (QLTT) on plasma calcitonin gene-related peptide (CGRP), beta-endorphin (β-EP), 5-HT, dopamine (DA), noradrenalin (NE), and blood viscosity in migraine model rats and mice.MethodsBoth the acute blood stasis model group and nitroglycerin-induced migraine model group included 60 Sprague-Dawley rats. The reserpine-reduced model group had 60 Kunming mice. Rats from each test were grouped into normal control group, model group, Zhengtian pill (ZTP) group, and high, moderate, or low-dose QLTT groups. In the acute blood stasis model test, after gavage for 7 days, rats were given 0.8 mL/kg adrenaline hydrochloride subcutaneously twice, and kept in ice water for 5 min. After fasting for 12 h, rats were anesthetized and blood samples were collected for detection of blood viscosity. In the nitroglycerin-induced migraine group, after gavage for 7 days, rats were intraperitoneally injected nitroglycerin (10 mg/kg), and 4 h later, blood samples were collected from postcava for measuring the plasma CGRP and β-EP levels. In the reserpine-reduced model test, except the normal control group, mice were administered reserpine (0.25 mg/kg, i.h.) for 9 days. Mice received intragastric administration from the third day to the ninth day. One hour after the last gavage, blood and brain tissue samples were obtained. Then, blood clotting time and the contents of neurotransmitters were determined.ResultsQLTT- (3.6, 1.8, and 0.9 g/kg) and ZTP-treated rats had lower blood viscosity than that in model rats under different shear rates (P< 0.01). QLTT- (3.6, 1.8 g/kg) and ZTP-treated rats had significantly lower plasma CGRP levels and higher plasma β-EP levels than those in model rats (P< 0.01). QLTT treatment at dose of 0.9 g/kg had lower plasma CGRP levels as well (P

Introduction Chromones and pyrones are the major secondary metabolites of Aloe barbadensis Miller. As they are minor components of the plant, an efficient purification procedure for them is of great importance for promoting their pharmacological studies. Objective To develop efficient methods for one-step separation and purification of two chromones (5-((S)-2′-oxo-4′-hydroxypentyl)-2-hydroxymethylchromone (1) and 5-((4E)-2′-oxo-pentenyl)-2-hydroxymethylchromone (3)) and one pyrone (aloenin aglycone (2)) from A. barbadensis via reversed-phase flash chromatography (RP-FC) and high-speed counter current chromatography (HSCCC). Methods The RP-FC separation was performed using methanol:water (26:74, v/v) as the mobile phase at a flow rate of 20 mL/min. A solvent system composed of dichloromethane:methanol:water (3:1.5:1, v/v/v) was used for the HSCCC separation, at a flow rate of 2.0 mL/min. Results A one-step RP-FC operation within 110 min was successfully used for the purification of compounds 1 (27.9 mg, 96.5%), 2 (32.4 mg, 98.2%) and 3 (4.1 mg, 99.0%) from 129 mg of crude sample, and a one-step HSCCC separation within 95 min was successfully implemented for the purification of compounds 1 (31.1 mg, 97.6%), 2 (35.8 mg, 96.7%) and 3 (2.7 mg, 98.1%) from 134 mg of crude sample. Conclusion The developed procedures were efficient, with low cost and high yield, which would afford sufficient amounts of high-purity compounds for chromatographic purposes and pharmacological activity screening. Copyright © 2014 John Wiley & Sons, Ltd.

Summary This study explored the stability and degradation products of aloin A under varying pH, temperature and light conditions usually encountered in processing. The stability of aloin A was significantly affected by temperature and pH. The content of aloin A decreased by more than 90% within 12 h at 50 °C and within 6 h at 70 °C, respectively. A significant decrease in stability was also observed at higher pHs. At pH 8.0, less than 2% of aloin A remained within 12 h. However, aloin A exhibited good stability at acidic pH levels with 94% remaining at pH 2.0 for 14 days. Light exerted no influence on the stability during the experimental period (14 days). Aloe-emodin, elgonica-dimers A and B were characterised as major degradation products of aloin A at pH 5.0 or below, and elgonica-dimers were mainly formed at 4 °C as well. 10-hydroxyaloins A and B were found under any condition except at pH 2.0 and 3.0, and they were mainly formed under high temperature, neutral-basic and any light conditions.

Aloe barbadensis Mill has been used as food and medicine for a long time. In order to investigate the chemical constituents of A. barbadensis and their inhibitory activities towards phosphodiesterase-4D (PDE4D), 70% methanol extract of the dried A. barbadensis powder was employed. Phytochemical investigation has led to the isolation of three new chromones, 5-(hydroxymethyl)-7-methoxy-2-methylchromone (4), 5-((4E)-2′-oxo-pentenyl)-2-hydroxymethylchromone (6), and 7-hydroxy-5-(hydroxymethyl)-2-methylchromone (7), together with eighteen known compounds. Their chemical structures were determined based on spectroscopic methods including UV, IR, 1D and 2D NMR, and HRMS spectrometry. In addition, their inhibition against PDE4D was evaluated using tritium-labeled adenosine 3′,5′-cyclic monophosphate (3H-cAMP) as the substrate. Inhibition was calculated by the variation of radioactivity after the reaction, and compounds 1–4, 10, and 21 exhibited certain inhibitory activities towards PDE4D, which can provide an explanation why A. barbadensis can serve as anti-inflammatory agents.

In the last three decades, China has experienced dual decentralization in transforming its economy, resulting in decentralization and deconcentration in public administration and social service delivery. Under the policy framework of decentralization, particularly when welfare financing has been decentralized to local governments, the emergence of welfare regionalism is evident in China. This paper sets out against this policy context to examine how three local governments in eastern coastal areas in China handle changing labour welfare needs by institutionalizing social and labour protection measures to meet the local needs instead of implementing central policy. With particular reference to examine why regional variations exist in welfare provision even though the socio-economic development status is similar in these areas, this paper shows how different forms of capitals, government's governance style and diversity of industries have affected welfare arrangements for labour in China.

To investigate the effect of low-selenium diet on the liver and kidneys of rats and explore the role of macrophage polarization into M1 and M2 phenotypes in liver and kidney injuries.Twenty-four rats (12 female and 12 male) were randomly divided into control group and low-selenium group and fed with normal chow (dietary selenium of 0.18 mg/kg) and low-selenium diet (dietary selenium of 0.02 mg/kg) for 109 days. After the feeding, the rats were sacrificed for HE staining to observe liver and kidney pathologies, and immunohistochemistry was performed for analyzing CCR7, CD206, CD163-positive cell numbers in the liver and kidneys.The rats in low-selenium group showed severer fibrosis in the liver and kidney than the control group. In either male or female rats in low-selenium group, CCR7 and CD206 expressions in the liver were comparable with those in control group, but CD163 expression was lower than that in the control group (P0.05 for both female and male rats). In the kidney, the proximal tubule showed a slightly higher while the distal tubule showed a slightly lower CCR7 expression in low selenium group than in the control group (P0.05). In low-selenium group, a significantly lower CD163 expression in the distal tubule and a significantly higher CD206 expression in the proximal tubule were noted as compared with the control group (P0.05 in both female and male rats). Compared with the control rats, the male rats in low-selenium group, but not the female rats, showed a significantly lower CD163 expression in the proximal tubule of the kidney (P0.05); the female but not the male rats in low-selenium group show a higher CD206 expression in the distal tubule (P0.05).Low-selenium diet can cause liver and kidney fibrosis in rats and may inhibit macrophage activation into the M2 phenotype.

Background The imbalance of Treg/Th17 cells is an important pathogenic factor for immune thrombocytopenic purpura (ITP). We previously reported miR-125a-5p targeted CXCL13 and participated in the process of ITP. In the present study, the role of miR-125a-5p in regulating Treg/Th17 ratio and its potential molecular mechanism were investigated. Method A total of 30 adults with ITP and 30 healthy subjects were included. MEG3 expression in peripheral blood derived CD4+ T cells from ITP patients and healthy subjects were detected by real-time PCR. In vitro experiments, the effects of inhibiting or overexpressing MEG3 on the expression of miR-125a-5p, Foxp3 and ROTγt in CD4+ T cells were investigated. Results MEG3 expression was increased in CD4+ T cells of patients with ITP. Dexamethasone decreased MEG3 expression level of CD4+ T cells in vitro . MEG3 directly interacted with miR-125a-5p and MEG3 overexpression inhibited miR-125a-5p expression in CD4+ T cells exposed to dexamethasone. MEG3 down-regulation or miR-125a-5p overexpression promoted Foxp3 expression and inhibited RORγt expression. Conclusion MEG3 interacted with miR-125a-5p and inhibited its expression, and MEG3/miR-125a-5p contributed to induce immune imbalance of Treg/Th17 in ITP.

Process capability assessment is important to statistical quality control. The existing univariate or multivariate process capability indices cannot be directly applied to assess the process for the quality characterized by a simple linear profile. There has been little attempt to study the process capability assessment in simple linear profile. This paper proposes a method to assess process capability for simple linear profile. Since two-dimensional predictions of the slope and the intercept of simple linear profile can represent the prediction profile, the process capability analysis can be treated as a two-dimensional correlated variables problem. Then, a multivariate process capability index based on a vector of three components is applied to assess the process capability in simple linear profile. The proposed method was proved by a simulation study.

Carbon emission permit trading is widely considered an effective means to achieve optimal distribution of regional environment. The initial allocation of carbon emissions is the bases of the successful implementation of industry carbon trading. Developing low carbon economy and achieving the state’s energy saving targets are effective ways to the adjustment of economic structure, the improvement of energy efficiency and the development of new industries. The initial allocation of carbon emissions trading has a significant impact on the carbon emissions trading policy. The paper provides a theoretical basis for the choice of the allocation methods of carbon emission permit trading through the research on the fairness of carbon emission trading.

The ethanolic extract of Aloe barbadensis Miller leaf skin showed inhibitory activity against phosphodiesterase-4D (PDE4D), which is a therapeutic target of inflammatory disease. Subsequent bioassay-guided fractionation led to the isolation of two new anthrones, 6'-O-acetyl-aloin B (9) and 6'-O-acetyl-aloin A (11), one new chromone, aloeresin K (8), together with thirteen known compounds. Their chemical structures were elucidated by spectroscopic methods including UV, IR, 1D and 2D NMR, and HRMS. All of the isolates were screened for their inhibitory activity against PDE4D using tritium-labeled adenosine 3',5'-cyclic monophosphate ((3)H-cAMP) as substrate. Compounds 13 and 14 were identified as PDE4D inhibitors, with their IC50 values of 9.25 and 4.42 μM, respectively. These achievements can provide evidences for the use of A. barbadensis leaf skin as functional feed additives for anti-inflammatory purpose.

A quality control strategy using high-performance liquid chromatography-diode array detector-electrospray ionization-tandem mass spectrometry (HPLC-DAD-ESI-MS/MS) coupled with chemometrics analysis was proposed for Aloe barbadensis Miller. Firstly, the extraction conditions including methanol concentration, extraction time and solvent-to-material ratio were optimized by multi-responses optimization based on response surface methodology (RSM). The optimum conditions were achieved by Derringer's desirability function and experimental validation implied that the established model exhibited favorable prediction ability. Then, HPLC fingerprint consisting of 27 common peaks was developed among 15 batches of A. barbadensis samples. 25 common peaks were identified using HPLC-DAD-ESI-MS/MS method by their spectral characteristics or comparison with the authentic standards. Chemometrics techniques including similarity analysis (SA), principal components analysis (PCA) and hierarchical clustering analysis (HCA) were implemented to classify A. barbadensis samples. The results demonstrated that all A. barbadensis samples shared similar chromatographic patterns as well as differences. These achievements provided an effective, reliable and comprehensive quality control method for A. barbadensis.

Objective To evaluate the analgesic and sedative effects of Qilong Toutong Granule (芪龙头痛颗粒, QTG) and explore its possible mechanisms.

The purpose of this study was to develop RT- PCR assay for Rapidly detect and distinguish between Norovirus genogroup I and genogroup II with a pair of primers.A pairs of primers specific to capsid prote in ORF2 gene of G I and G II Norovirus were dsigned according to the published complete genome sequence, with which the RNA of Norovirus was extracted and RT-PCR amplification. The sensitivity, specificity of the RT- PCR assay was estimated and apply it to the detection of Norovirus in clinical specimens.The results showed that the assay possessed high specificity for Norovirus detection and without any evident cross-reaction with other viruses, including rotavirus, enteric adenovirus and hepatitis A virus. The detection limit of RT-PCR assay for Norovirus G I and G II were up to 100 pg/ml and 10 pg/ml respectively.The RT- PCR assay provide rapid and sensitive detection of Norovirus G I and G II and should prove to be useful for Norovirus diagnosis in the outbreaks of acute gastroenteritis.

Chromones and pyrones are the major secondary metabolites of Aloe barbadensis Miller. As they are minor components of the plant, an efficient purification procedure for them is of great importance for promoting their pharmacological studies.To develop efficient methods for one-step separation and purification of two chromones (5-((S)-2'-oxo-4'-hydroxypentyl)-2-hydroxymethylchromone (1) and 5-((4E)-2'-oxo-pentenyl)-2-hydroxymethylchromone (3)) and one pyrone (aloenin aglycone (2)) from A. barbadensis via reversed-phase flash chromatography (RP-FC) and high-speed counter current chromatography (HSCCC).The RP-FC separation was performed using methanol:water (26:74, v/v) as the mobile phase at a flow rate of 20 mL/min. A solvent system composed of dichloromethane:methanol:water (3:1.5:1, v/v/v) was used for the HSCCC separation, at a flow rate of 2.0 mL/min.A one-step RP-FC operation within 110 min was successfully used for the purification of compounds 1 (27.9 mg, 96.5%), 2 (32.4 mg, 98.2%) and 3 (4.1 mg, 99.0%) from 129 mg of crude sample, and a one-step HSCCC separation within 95 min was successfully implemented for the purification of compounds 1 (31.1 mg, 97.6%), 2 (35.8 mg, 96.7%) and 3 (2.7 mg, 98.1%) from 134 mg of crude sample.The developed procedures were efficient, with low cost and high yield, which would afford sufficient amounts of high-purity compounds for chromatographic purposes and pharmacological activity screening.

To investigate the chemical constituents of A. barbadensis, aqueous extract of the plant was subjected to preparative medium pressure liquid chromatography (MPLC). The chemical structures were mainly determined by spectroscopic evidences (UV, IR, HR-MS, 1H NMR, 13C NMR, HSQC, 1H-1H COSY and HMBC) and chemical methods. A new O, O, O-triglucosylated naphthalene derivative, together with two known 6-phenyl-2-pyrone derivatives and four 5-methylchromones, were isolated and identified as 1-((3-((4- O-beta-D-glucopyranosyl)-beta-D-xylopyranosyloxymethyl)-1-hydroxy-8-alpha-L-rhamnopyranosyloxy)naphthalene-2-y])-ethanone (1), 10-O-beta-D-glucopyranosyl aloenin (2), aloenin B (3), aloesin (4), 8-C-glucosyl-(R)-aloesol (5), 8-C-glucosyl-7-O-methyl-(S)-aloesol (6), and isoaloeresin D (7). Compound 1 is a novel naphthalene derivative and named as aloveroside B, compounds 2-3 are isolated from this Aloe species for the first time.

To evaluate the analgesic and sedative effects of Qilong Toutong Granule (, QTG) and explore its possible mechanisms.Kunming mice were randomly divided into 6 groups: normal control group, Zhengtian Pill (, ZTP) group, Western medicine group, and high-dose (5.2 g/kg), medium-dose (2.6 g/kg) and low-dose (1.3 g/kg) of QTG groups. After completing the prophylactic treatment for 3 days, hot-plate test and acetic acid-induced writhing test were used to assess the analgesic effect, and spontaneous locomotor test and sodium pentobarbital-induced hypnosis activity were adopted to estimate the sedative effect. Sprague-Dawley rats were grouped into normal control group, model group, ZTP group, rizatriptan group, and high-dose (3.6 g/kg), medium-dose (1.8 g/kg), and low-dose (0.9 g/kg) of QTG groups. After gavage for continuous 7 days, rats were intraperitoneally injected nitroglycerin, and 4 h later, blood samples were collected from postcava for measuring the levels of plasma calcitonin gene-related peptide (CGRP) and beta-endorphin (β-EP) by radioimmunoassay. Subsequently, rats were perfused transcardially and the brain tissues containing the trigeminal nucleus caudalis (TNC) were achieved for detecting the number of Fos-immunoreactive cells by immunohistochemical method.In the mice experiments, compared with the normal control group, high- and medium-dose of QTG groups significantly raised the pain threshold (P0.01), reduced the number of writhing response (P0.01) and spontaneous activity (P0.01), but had no influence on the sleeping rate of mice (P0.05), and low-dose of QTG group also raised the pain threshold at 120 min (P=0.007), as well as lowered locomotor activity of mice at 2 h (P=0.003). On the study of migraine model rats, high- and medium-dose of QTG groups remarkably down-regulated the levels of plasma CGRP (P0.01), up-regulated the levels of plasma β-EP (P0.01) and inhibited the expression of Fos protein in TNC (P0.01), compared with the model group.QTG has obvious analgesic and sedative action and its mechanism on relieving migraine may be through regulating the levels of neurotransmitters and/or neuropeptides, and inhibiting the activation of Fos pathway.

To study the molecular characteristics of Noroviruses causing outbreaks of acute gastroenteritis in Huzhou.From 2008 to 2010, total 119 fecal specimens collected from outbreaks of acute gastroenteritis were tested for Norovirus. Partial sequence of RNA dependent RNA polymerase (RdRp) of the positive samples were amplified by RT-PCR, then the PCR production were purified, sequenced and put into phylogenetic analysis.50 of 119 specimens were positive for Norovirus by real-time RT-PCR. Out of those 50 Norovirus positive specimens, 9 were Norovirus Genogroup I (GI) positive, 35 were Norovirus Genogroup II (GII) positive, 6 was both Norovirus GI and GII positive. 12 PCR products for RdRp were selected for further studies on sequencing. Phylogenetic analysis revealed that the 5 GI norovirus isolates were belonged to genotype GI/2 and GI/3. Of the 7 GII norovirus isolates, 6 were belonged to genotype GII/4, 1 was belonged to genotype Glib.Norovirus is a major cause of outbreaks of acute gastroenteritis in Huzhou and the epidemic strains of norovirus isolated from Huzhou had a high degree of genetic diversity.

To study the molecular characteristics of Noroviruses causing outbreaks of acute gastroenteritis in Huzhou. During April 2008 and February 2009, fecal specimens of patients collected from 2 outbreaks of acute gastroenteritis were tested for Norovirus by real-time RT-PCR. Partial sequence of RNA dependent RNA polymerase(RdRp) of the positive samples were amplified by RT-PCR, the PCR products were then purified, sequenced and phylogenetic analysis was conducted. Both genogroup II (GII) and genogroup I (GI) noroviruses were detected in 2 outbreaks. Phylogenetic analysis revealed that two of the GI norovirus strains isolated from 2008 belonged to genotype GI/2 and one of the GI Norovirus strain isolated from 2009 belonged to genotype GI/3. The other GIIú norovirus strains isolated from 2009 had high nucleotide identity with GIIb genotype that had been reported frequently in European countries during 2000 and 2001 and in Asian countries recently. These results suggested that the epidemic strains of norovirus isolated in Huzhou had a high degree of genetic diversity and prevalent genotypes at different times were also different. To our knowledge this is the first report of detecting GIIb variant in outbreaks of acute gastroenteritis in China.

Taguchi method is the most popular approach for multiresponse optimization. However, the weighting values of responses are ignored in this approach. Thus improved Taguchi method is presented, which applies the principal component analysis to evaluate the weights so that their relative importance can be properly described. The implementation and effectiveness of the proposed method are illustrated by an example from the literature. The results show that improved Taguchi method can optimize the multiresponses.

To investigate the expression of rat protamine (RP) gene in MEL cells and the effect on cell growth.Eukaryotic expression plasmid pCR-3.1-RP was constructed and transfected into MEL cells. The changes of cell growth rate, mitotic index and colony-forming rate in semi-solid medium were investigated.Transfected MEL cells showed lower growth rate, mitotic index and colony-forming rate. Volumes of cells were reduced and reduction of RNA transcription was observed.These results suggest that expression of RP in MEL cell may inhibit the cell growth and proliferation.

Multi-component analysis has been considered to be one of the key techniques for quality control of traditional Chinese medicines. However, the shortage and high cost of reference standards have become the greatest obstacles in implementing this goal. An alternative method using only an easily available reference standard as the internal standard to determine multiple analogues has gained increasing attention. In the present study, a practical HPLC method was established for the simultaneous quantification of seven chromones in Aloe barbadensis Miller using a single reference standard. An easily available compound in the plant named 8-C-glucosyl-7-O-methyl-(S)-aloesol (3) was selected as the internal standard, and the other six chromones were determined by their relative response factors. The results indicated that the established method was accurate and reliable for the routine quality control of A. barbadensis. 6 batches of A. barbadensis samples were analyzed using this method and no significant difference was observed in the results obtained using the established method and external standard method. The newly developed method can serve as an alternative approach when reference standards are difficult to obtain. This achievement would promote the quality control of A. barbadensis.