Two new sesterterpenoids from Atractylodes japonica Koidz. ex Kitam.

Two new sesterterpenoids, atractylodes japonica terpenoid acid I (1) and atractylodes japonica terpenoid aldehyde I (2), were isolated from the rhizomes of Atractylodes japonica Koidz. ex Kitam together with ten known compounds (3-12). Their structures were elucidated on the basis of comprehensive spectroscopic analysis (1D/2D NMR, HRESIMS and IR). In addition, all of these isolated compounds were evaluated for their cytotoxic activities against human gastric cancer cell MGC-803 and human hepatocellular cancer cell HepG-2. Most of them exhibited moderate to weak inhibitory effects with IC50 values in the range of 25.15-88.85 µM except for 9-12.

Highly selective butanol production by manipulating electron flow via cathodic electro-fermentation.

Butanol production by solventogenic Clostridia shows great potential to combat the energy crisis, but is still challenged by low butanol selectivity and high downstream cost. In this study, a novel cathodic electro-fermentation (CEF) system mediated by methyl viologen (MV) was proposed and sequentially optimized to obtain highly selective butanol production. Under the optimal conditions (-0.60 V cathode potential, 0.50 mM MV, 30 g/L glucose), 7.17 ± 0.55 g/L butanol production were achieved with the yield of 0.32 ± 0.02 g/g. With the supplement of 4 g/L butyric acid as co-substrate, butanol production further improved to 13.14 ± 1.14 g/L with butanol yield and selectivity as high as 0.43 ± 0.01 g/g and 90.44 ± 1.66%, respectively. The polarized electrode enabled the unbalanced fermentation towards butanol formation and MV further inhibited hydrogen production, both of which contributed to the high-level butanol production and selectivity. The MV-mediated CEF system is a promising approach for cost-effective bio-butanol production.

Extensive crystal fractionation of high-silica magmas revealed by K isotopes.

Fractional crystallization plays a critical role in generating the differentiated continental crust on Earth. However, whether efficient crystal-melt separation can occur in viscous felsic magmas remains a long-standing debate because of the difficulty in discriminating between differentiated melts and complementary cumulates. Here, we found large (~1 per mil) potassium isotopic variation in 54 strongly peraluminous high-silica (silicon dioxide >70 weight %) leucogranites from the Himalayan orogen, with potassium isotopes correlated with trace elemental proxies (e.g., strontium, rubidium/strontium, and europium anomaly) for plagioclase crystallization. Quantitative modeling requires up to ~60 to 90% fractional crystallization to account for the progressively light potassium isotopic composition of the fractionated leucogranites, while plagioclase accumulation results in enrichment of heavy potassium isotopes in cumulate leucogranites. Our findings strongly support fractional crystallization of high-silica magmas and highlight the great potential of potassium isotopes in studying felsic magma differentiation.

Effect of Roucongrong (Herba Cistanches Deserticolae) decoction on the substantia nigra through Wnt/ß-catenin signaling pathway in rats with Parkinson's disease induced by 6-hydroxydopamine hydrochloride.

OBJECTIVE: To investigate the effect of Roucongrong (Herba Cistanches Deserticolae) decoction on the substantia nigra in rats with Parkinson's disease (PD) induced by 6-hydroxydopamine hydrochloride (6-OHDA). To further determine whether the Wnt/ß-catenin signaling pathway is involved in the action. METHODS: A rat model of PD was established by intracranial injection of 6-OHDA. Subsequently, three concentrations of Roucongrong (Herba Cistanches Deserticolae) decoction were prepared and administered to rats by gavage therapy for 14 d. Behavioral changes were measured in PD rats. In vivo tyrosine hydroxylase (TH) levels in the substantia nigra were examined by immunohistochemistry. Additionally, gene and protein expression levels of members of the Wnt/ß-catenin signaling pathway were examined by Western blotting and polymerase chain reaction. Lastly, a Wnt/ß-catenin inhibitor was used to investigate the mechanism of action in 1-methyl-4-phenylpyridinium (MPP + )- treated MES23.5 cells in vitro. RESULTS: Roucongrong (Herba Cistanches Deserticolae) decoction improved performance in the stride and gait adjustment tests in PD rats. It also increased TH in the substantia nigra and altered the expression of genes and proteins in the Wnt/ß-catenin signaling pathway. Wnt/ß-catenin inhibitor reduced the effect of Roucongrong (Herba Cistanches Deserticolae) decoction in MPP +-treated MES23.5 cells. CONCLUSION: Roucongrong (Herba Cistanches Deserticolae) decoction may promote neuronal survival in PD in vivo and in vitro by increasing TH content in the substantia nigra and by activating the Wnt/ß-catenin signaling pathway.

Discovery of a potent and selective inhibitor of histone lysine demethylase KDM4D.

Histone lysine demethylase 4D (KDM4D) plays an important role in the regulation of tumorigenesis, progression and drug resistance and has been considered a potential target for cancer treatment. However, there is still a lack of potent and selective KDM4D inhibitors. In this investigation, we report a new class of KDM4D inhibitors containing the 2-(aryl(pyrrolidine-1-yl)methyl)phenol scaffold, identified through AlphaLisa-based screening, structural optimization, and structure-activity relationship analyses. Among these inhibitors, 24s was the most potent, with an IC50 value of 0.023 ± 0.004 µM. This compound exhibited more than 1500-fold selectivity towards KDM4D versus KDM4A as well as other JMJD subfamily members, indicating good selectivity for KDM4D. Kinetic analysis indicated that 24s did not occupy the 2-oxoglutarate binding pocket. In an in vitro assay, 24s significantly suppressed the proliferation and migration of colorectal cancer (CRC) cells. Overall, this study has identified a good tool compound to explore the biological function of KDM4D and a good lead compound for drug discovery targeting KDM4D.

Neolignan and phenylpropanoid compounds from the resin of Styrax tonkinensis.

A new neolignan compound, tonkinensisin A (1), and two phenylpropanoid compounds, tonkinensisin B (2) and tonkinensisin C (3), were isolated from the resin of Styrax tonkinensis (Pierre) Craib ex Hartw. The structures of these compounds were determined by spectroscopic analysis, and the relative configurations of compounds 1 and 3 were determined by J-based configuration analysis (JBCA) method. All compounds were assayed for cytotoxic activities against five tumor cell lines (HepG-2, A549, Hela, MCF-7, and PC-3) by CCK-8 test in vitro.[Formula: see text].

Dihydromyricetin reverses MRP2-induced multidrug resistance by preventing NF-κB-Nrf2 signaling in colorectal cancer cell.

BACKGROUD: Dihydromyricetin (DMY), a natural flavonoid compound from the leaves of the Chinese medicinal herb Vitis heyneana, has been shown to have the potential to combat chemoresistance by inhibiting Nrf2/MRP2 signaling in colorectal cancer (CRC) cells. However, the precise underlying molecular mechanism and its therapeutic target are not well understood. PURPOSE: Our study aims to investigate the effects of DMY on multidrug resistance (MDR), and elucidate the underlying mechanisms. STUDY DESIGN: In vitro, HCT116/OXA and HCT8/VCR cells were employed as our MDR models. The cells were treated with DMY (50 µM) or MK-571 (50 µM) plus oxaliplatin (OXA) (10 µM) or vincristine (VCR) (10 µM) for 48 h. In vivo, we used BALB/c mice as a CRC xenograft mouse model. BALB/c mice were given DMY (100 mg/kg), OXA (5 mg/kg) and DMY (100 mg/kg) combined with OXA (5 mg/kg) via intraperitoneal route every 2 days per week for 4 weeks. METHODS: We used MTT and colony forming assays to detect DMY's ability to reverse MDR. Flow cytometric analysis was used to detect apoptosis. Immunocytochemistry was used to detect the localization of Nrf2 and NF-κB/p65. Western blot, qRT-PCR and reporter gene assays were employed to measure the protein and gene transcriptional levels (MRP2, Nrf2, NF-κB/p65). Moreover, chromatin immunoprecipitation (ChIP) assay was used to investigate the endogenous promoter occupancy of NF-κB/p65. Finally, immunohistochemistry and TUNEL staining were used to detect protein expression and apoptosis in vivo. RESULTS: DMY restored chemosensitivity (OXA and VCR) by inhibiting both MRP2 expression and its promoter activity in HCT116/OXA and HCT8/VCR cell lines. Furthermore, DMY could inhibit NF-κB/p65 expression, reducing NF-κB/p65 translocation to the nucleus to silence Nrf2 signaling, which is necessary for MRP2 expression. Overexpressing NF-κB/p65 expression reduced the reversal effect of DMY. In addition, NF-κB/p65 regulated Nrf2 expression by directly binding to its specific promoter region and activating its transcription. Finally, we proved that the combination of OXA and DMY has a synergistic tumor suppression effect in vivo. CONCLUSION: Our study provided a novel mechanism of DMY boosted chemosensitivity in human CRC. The downstream signals of DMY, NF-κB or Nrf2 could also be potential targets for the treatment of CRC.

Cysestermerol A, a rare stilbene sestermer with significant hypoglycemic activities from Cynodon dactylon.

Cysestermerol A (1), a rare and new stilbene sestermer, was isolated from the whole herb of Cynodon dactylon. The planar and relative structures of 1 were elucidated based on HRESIMS, one- and two-dimensional NMR analyses, and its absolute configuration was further established by electronic circular dichroism calculations. Compound 1 obviously increased the glucose consumption in HepG2 cells equivalent to the positive control rosiglitazone and markedly inhibited the activity of α-glucosidase in vitro.

Green synthesis of Ag and Pd nanoparticles for water pollutants treatment.

Silver (Ag) and palladium (Pd) nanoparticles were synthesized via a green synthesis route, which was mediated with the extract of Daucus carota leaves. The morphological, crystalline and structural nature of the synthesized nanoparticles was characterized by UV-Vis spectrophotometer, and TEM, XRD and FT-IR analyses. High antibacterial activities of the prepared Ag and Pd nanoparticles were observed towards different water-borne pathogens of Klebsiella pneumonia, Vibrio cholera and Escherichia coli. The catalytic efficiency of the prepared nanoparticles for the removal of rhodamine 6G (Rh-6G) dye was also evaluated. Nearly 98% of the Rh-6G dye was decolorized by the synthesized Pd nanoparticles within 2 min, and the synthesized Ag nanoparticles took 30 min for 89.4% decolorization. This work provided greener nanocatalysts for pollutant treatment and demonstrated the power of green biosynthesis for metallic nanoparticles.

[Effects of repeated immobilization stress on hypothalamic-pituitary- ovarian axis in female rats].

Objective: To explore the effects of repeated immobilization stress on hypothalamic-pituitary-ovarian axis in female rats. Methods: Forty female SD rats were randomly divided into two groups: control group (n=20) and experimental group (n=20). One group was fed normally, the other group was subjected to incremental load restraint stress. Brake stress once a day in the retainer (starting at 9: 00 a.m.), braking for 2 hours on the first day, increasing load by 0.5 hours a day for two weeks. Body weight, estrous cycle, sex hormone, organ coefficient, pathology and expression of related genes were detected to explore the harm of hypothalamic-pituitary-ovarian axis. Results: Repeated immobilization stress caused weight loss, prolonged estrous cycle, and changed the organ coefficient and morphology of ovaries and uterus. QPCR technique was used to detect the related genes. It was found that the expressions of gonadotropin releasing hormone, pituitary gonadotropin releasing hormone receptor, follicle stimulating hormone and luteinizing hormone mRNA were decreased significantly, while the expressions of ovarian follicle stimulating hormone and luteinizing hormone receptor mRNA were increased significantly. The expression of estrogen receptor mRNA in ovary and uterus was decreased significantly. Conclusion: Repeated immobilization stress may disrupt the estrous cycle by interfering with the endocrine regulation of the hypothalamic-pituitary-ovarian axis, thus damaging the gonadal and reproductive endocrine function of female animals.

Comparison and optimization of extraction protocol for intracellular phosphorus and its polyphosphate in enhanced biological phosphorus removal (EBPR) sludge.

Intracellular polyphosphate (poly-P) plays important roles in Enhanced biological phosphorus removal (EBPR) process, but an effective and reliable protocol for extracting intracellular P and its poly-P in EBPR sludge without hydrolysis of poly-P has not been setup yet. In the study, it was revealed that the severe hydrolysis of intracellular poly-P occurred during the different extraction processes, such as acid (i.e., HClO4, H2SO4 and HCl), basic (i.e., NaOH and KOH) and freezing-grind (under different solid-liquid ratios), but it did not occur during ultrasonic extraction process. The optimal extraction process of the ultrasonic protocol was 10 w/mL of ultrasonic power density and 15 min of ultrasonic time, when the extraction efficiency of intracellular P was 88.24 ±â€¯1.56%. In addition, the extraction efficiency of intracellular P could be furtherly improved by that the 0.75 mol/L LiCl solution was used to resuspend the bacterial cell before ultrasonic extraction (i.e., LiCl-ultrasonic protocol). The ultrasonic protocol was more suitable to extract the intracellular P and its poly-P of EBPR sludge than the other 4 protocols (i.e., PCA-NaOH, EDTA-NaOH, freezing-grind and LiCl-ultrasonic), which had the technical characteristics of (i) with relatively high extraction efficiency of intracellular P, (ii) without hydrolysis of intracellular poly-P, (iii) with weak noise signal in 31P NMR spectrum and (iv) with simple extraction process and short extraction time. It was founded by the ultrasonic protocol that there was the high content (82.88%-89.79% of intracellular P content) of intracellular poly-P with long average chain length (376.4-383.2) in the EBPR sludges. Importantly, it was confirmed that the EBPR process was related to the combined action of extracellular and intracellular poly-P using a new fractionation method of P in EBPR sludge, which included the ultrasonic protocol at high power density for extracting the intracellular P and its poly-P.

Novel sesquilignan and lignan glycoside from the twigs and leaves of Illicium majus.

A novel sesquilignan compound (1), possesing an unusual carbon skeleton of aryltetralin unit linked with a C6-C3 unit and a five-membered ring by a C-7″ and C-4 linkage pattern via an oxygen atom, and a new lignan glycoside (2) have been isolated from the twigs and leaves of Illicium majus. Their structures were determined by spectroscopic analysis and chemical methods. The absolute configurations of 1 and 2 are confirmed by observing the circular dichroism. At 10 µM, Compounds 1 and 2 showed in Vitro inhibitory activity against the release of ß-glucuronidase in rat polymorphonuclear leukocytes induced by platelet-activating factor.

Discovery of pyrazolo[1,5-a]pyrimidine-3-carbonitrile derivatives as a new class of histone lysine demethylase 4D (KDM4D) inhibitors.

Herein we report the discovery of a series of new small molecule inhibitors of histone lysine demethylase 4D (KDM4D). Molecular docking was first performed to screen for new KDM4D inhibitors from various chemical databases. Two hit compounds were retrieved. Further structural optimization and structure-activity relationship (SAR) analysis were carried out to the more selective one, compound 2, which led to the discovery of several new KDM4D inhibitors. Among them, compound 10r is the most potent one with an IC50 value of 0.41±0.03µM against KDM4D. Overall, compound 10r could be taken as a good lead compound for further studies.

[Acupuncture combined with western medication for mild cognitive disorder after stroke: a rando- mized controlled trial].

OBJECTIVE: To verify the clinical effect of the penetration acupuncture at the acupoints of governor vessel(GV) and bladder meridian on head for mild cognitive disorder after stroke. METHODS: One hundred and ten patients were randomly divided into an observation group and a control group, 55 cases in each one. Penetration acupuncture was used at the acupoints of GV and bladder meridian on head in the observation group,namely Wuchu (BL 5) to Chengguang (BL 6) and Tongtian (BL 7) to Luoque (BL 8) bilaterally, and Shenting (GV 24) to Xinhui (GV 22), Qianding (GV 21) to Baihui (GV 20), Houding (GV 19) to Qiangjian (GV 18). Body acupuncture and conventional western medication were applied in the both two groups. Acupuncture was adopted once a day, five times a week and eight weeks totally. The changes of Montreal Cognitive Assessment (MoCA), Mini mental state examination scale(MMSE), self-rating depressive scale (SDS) and activities of daily living scale (Barthel index) were compared between the two groups before treatment, in the middle of treatment(at the forth week) after treatment(at the eighth week) and in three weeks after treatment(follow-up). RESULTS: After treatment of four and eight weeks and at the time of follow-up, the results of MoCA, MMSE, SDS and Barthel index were apparently im proved compared with those before treatment in the two groups (all P < 0.01). After treatment of eight weeks and at the time of follow-up, the scores of MoCA and SDS were obviously improved in the observation group compared with those in the control group (P < 0.01, P < 0.05). The improvements of MMSE in the observation group were more marked than those in the control group at all times (P < 0.01, P < 0.05). But there was no statistical significance about the change of Barthel index between the two groups (all P > 0.05). CONCLUSION: Penetration acupuncture at the acupoints of GV and bladder meridian on head can remarkablely improve the cognitive function of the patients with mild cognitive disorder after stroke,and have some efficacy of relieving the bad mood such as depression.

Rhizoma Dioscoreae extract protects against alveolar bone loss by regulating the cell cycle: A predictive study based on the protein­protein interaction network.

Rhizoma Dioscoreae extract (RDE) exhibits a protective effect on alveolar bone loss in ovariectomized (OVX) rats. The aim of this study was to predict the pathways or targets that are regulated by RDE, by re­assessing our previously reported data and conducting a protein­protein interaction (PPI) network analysis. In total, 383 differentially expressed genes (≥3­fold) between alveolar bone samples from the RDE and OVX group rats were identified, and a PPI network was constructed based on these genes. Furthermore, four molecular clusters (A­D) in the PPI network with the smallest P­values were detected by molecular complex detection (MCODE) algorithm. Using Database for Annotation, Visualization and Integrated Discovery (DAVID) and Ingenuity Pathway Analysis (IPA) tools, two molecular clusters (A and B) were enriched for biological process in Gene Ontology (GO). Only cluster A was associated with biological pathways in the IPA database. GO and pathway analysis results showed that cluster A, associated with cell cycle regulation, was the most important molecular cluster in the PPI network. In addition, cyclin­dependent kinase 1 (CDK1) may be a key molecule achieving the cell­cycle­regulatory function of cluster A. From the PPI network analysis, it was predicted that delayed cell cycle progression in excessive alveolar bone remodeling via downregulation of CDK1 may be another mechanism underling the anti­osteopenic effect of RDE on alveolar bone.

Acupuncture treatment for ischaemic stroke in young adults: protocol for a randomised, sham-controlled clinical trial.

INTRODUCTION: Stroke in young adults is not uncommon. Although the overall incidence of stroke has been recently declining, the incidence of stroke in young adults is increasing. Traditional vascular risk factors are the main cause of young ischaemic stroke. Acupuncture has been shown to benefit stroke rehabilitation and ameliorate the risk factors for stroke. The aims of this study were to determine whether acupuncture treatment will be effective in improving the activities of daily living (ADL), motor function and quality of life (QOL) in patients of young ischaemic stroke, and in preventing stroke recurrence by controlling blood pressure, lipids and body weight. METHODS AND ANALYSIS: In this randomised, sham-controlled, participant-blinded and assessor-blinded clinical trial, 120 patients between 18 and 45 years of age with a recent (within 1 month) ischaemic stroke will be randomised for an 8-week acupuncture or sham acupuncture treatment. The primary outcome will be the Barthel Index for ADL. The secondary outcomes will include the Fugl-Meyer Assessment for motor function; the World Health Organization Quality of Life BREF (WHOQOL-BREF) for QOL; and risk factors that are measured by ambulatory blood pressure, the fasting serum lipid, body mass index and waist circumference. Incidence of adverse events and long-term mortality and recurrence rate during a 10-year and 30-year follow-up will also be investigated. ETHICS AND DISSEMINATION: Ethics approval was obtained from the Ethics Committee of The Third Affiliated Hospital of Zhejiang Chinese Medical University. Protocol V.3 was approved in June 2013. The results will be disseminated in a peer-reviewed journal and presented at international congresses. The results will also be disseminated to patients by telephone during follow-up calls enquiring on the patient's post-study health status. TRIAL REGISTRATION NUMBER: ChiCTR-TRC- 13003317; Pre-results.

Catheter Ablation of Fascicular Ventricular Tachycardia: Long-Term Clinical Outcomes and Mechanisms of Recurrence.

BACKGROUND: Fascicular ventricular tachycardia (FVT) is a common form of sustained idiopathic left ventricular tachycardia with an Asian preponderance. This study aimed to prospectively investigate long-term clinical outcomes of patients undergoing ablation of FVT and identify predictors of arrhythmia recurrence. METHODS AND RESULTS: Consecutive patients undergoing FVT ablation at a single tertiary center were enrolled. Activation mapping was performed to identify the earliest presystolic Purkinje potential during FVT that was targeted by radiofrequency ablation. Follow-up with clinic visits, ECG, and Holter monitoring was performed at least every 6 months. A total of 120 consecutive patients (mean age, 29.3±12.7 years; 82% men; all patients with normal ejection fraction) were enrolled. FVT involved left posterior fascicle and left anterior fascicle in 118 and 2 subjects, respectively. VT was noninducible in 3 patients, and ablation was acutely successful in 117 patients. With a median follow-up of 55.7 months, VT of a similar ECG morphology recurred in 17 patients, and repeat procedure confirmed FVT recurrence involving the same fascicle. Shorter VT cycle length was the only significant predictor of FVT recurrence (P=0.03). Six other patients developed new-onset upper septal FVT that was successfully ablated. CONCLUSIONS: Ablation of FVT guided by activation mapping is associated with a single procedural success rate without the use of antiarrhythmic drugs of 80.3%. Arrhythmia recurrences after an initially successful ablation were caused by recurrent FVT involving the same fascicle in two thirds of patients or new onset of upper septal FVT in the remainder.

One-step production of biodiesel from oils with high acid value by activated Mg-Al hydrotalcite nanoparticles.

Activated Mg-Al hydrotalcite (HT-Ca) nanoparticles (<45 nm) were synthesized by co-precipitation and hydrothermal activation with aqueous Ca(OH)2 solution. They were characterized by various techniques including X-ray diffraction, inductively coupled plasma atomic-emission spectrometer, Brunauer-Emmett-Teller method, scanning electronic microscope-X-ray energy dispersive analysis and temperature programmed desorption method. HT-Ca presented both acidic and basic due to the formation of Mg4Al2(OH)14 · 3H2O, Mg2Al(OH)7 and AlO(OH) nanocrystals to esterify and transesterify oils with high acid value (AV). Under conditions of 5 wt% HT-Ca, 160 °C, 30/1 methanol/oil molar ratio and 4h, 93.4% Jatropha biodiesel yield was obtained at AV of 6.3 mg KOH/g with 4 cycles (biodiesel yield>86%). It was further found that it can resist free fatty acids, and biodiesel yield reached 92.9% from soybean oil with high AV of 12.1. HT-Ca catalyst showed a potential practical application for direct production of biodiesel from oils with high AV without pretreatment.

Chemical and biological insights into uranium-induced apoptosis of rat hepatic cell line.

Uranium release into the environment is a threat to human health, and the mechanisms of cytotoxicity caused by uranium are not well-understood. To improve our understanding in this respect, we herein evaluated the effects of uranium exposure on normal rat hepatic BRL cells. As revealed by scanning electron microscopy and transmission electron microscope analysis, uranyl nitrate was found to be transformed into uranyl phosphate particles in the medium and taken up by BRL cells in an endocytotic uptake manner, which presumably initiates apoptosis of the cell, although soluble uranyl ion may also be toxic. The apoptosis of BRL cells upon uranium exposure was also confirmed by both the acridine orange and ethidium bromide double staining assay and the Annexin V/propidium iodide double staining assay. Further studies revealed that uranium induced the loss of mitochondrial membrane potential in a dose-dependent manner. Moreover, the uranium-induced apoptosis was found to be associated with the activation of caspase-3, caspase-8 and caspase-9, indicating both a mitochondria-dependent signaling pathway and a death receptor pathway by a crosstalk. This study provides new chemical and biological insights into the mechanism of uranium toxicity toward hepatic cells, which will help seek approaches for biological remediation of uranium.