Quantitative analysis of anthocyanins and flavonoids in purple and green leaves of Eleutherococcus senticosus by UPLC-MS/MS techniques.

Ultra performance liquid chromatography-high resolution mass spectrometry (UPLC-MS/MS) were used to target quantitative determination anthocyanins and flavonoids in the fresh leaves (purple and green) of Eleutherococcus senticosus. The results showed that the content of total anthocyanins was 99.68 µg/g (Fresh Weight, FW) in purple leaves and 29.12 µg/g in green leaves. Cyanidin-3-O-galactoside and delphinidin were the main anthocyanins compound in purple and green leaves, and the content of the both declined sharply in green leaves. The content of cyanidin-3-O-galactoside reached 616.23 ng/100 mg in purple leaves and was only fifth in green leaves. The total flavonoids content was 4.90 mg/g in purple leaves and 2.23 mg/g in green leaves. Quercetin-3-ß-D-glucoside (236.96 ng/mg) and kaempferol-3-O-glucoside (145.27 ng/mg) were the main flavonoids compound in purple leaves. Besides the two main flavonoids, large quantities of rutin (269.11 ng/mg) was detected in green leaves of E. senticosus.

Magnetic and Ferroelectric Manipulation of Valley Physics in Janus Piezoelectric Materials.

The realization of multiferroic materials offers the possibility of multifunctional electronic device design. However, the coupling between the multiferroicity and piezoelectricity in Janus materials is rarely reported. In this study, we propose a mechanism for manipulating valley physics by magnetization reversing and ferroelectric switching in multiferroic and piezoelectric material. The ferromagnetic VSiGeP4 monolayer exhibits a large valley polarization up to 100 meV, which can be effectively operated by reversing magnetization. Interestingly, the antiferromagnetic VSiGeP4 bilayers with AB and BA stacking configurations allow the coexistence of valley polarization and ferroelectricity, supporting the proposed strategy for manipulating valley physics via ferroelectric switching and interlayer sliding. In addition, the VSiGeP4 monolayer contains remarkable tunable piezoelectricity regulated by electron correlation U. This study proposes a feasible idea for regulating valley polarization and a general design idea for multifunctional devices with multiferroic and piezoelectric properties, facilitating the miniaturization and integration of nanodevices.

Active Asymmetric Electron-Transfer Effect on the Enhanced Piezoelectricity in MoTO (T = S, Se, or Te) Monolayers and Bilayers.

Development of piezoelectric materials is limited partly due to the incompleteness of internal mechanism and the lack of vertical piezoelectricity. Herein, we theoretically identify the stable MoTO (T = S, Se, or Te) monolayers and bilayers. When two elements are given but another element can be changed, the larger the electronegativity difference ratio Rratio is, the stronger the piezoelectricity will be. Vertical piezoelectric coefficient d33 of the MoTeO bilayer reaches 38.907 pm/V, which is 12 times larger than that of the bulk GaN. The "active asymmetric electron-transfer" strategy mainly contributes to the spontaneous remarkable piezoelectricity of MoTO. Importantly, we proposed the new method for calculating the piezoelectric coefficients of two-dimensional (2D) materials, which corresponds to the fact that 2D materials have a certain thickness. This study not only provides novel extraordinary candidates for energy conversion and touch-sensor nanodevices but also promotes a deeper understanding of piezoelectricity of 2D materials.

[Polarized activation affects iron metabolism in macrophages].

The aim of this study was to investigate the effects of polarization program on the ability of macrophages to regulate iron metabolism. M1 and M2 macrophages were propagated in vitro from porcine alveolar macrophages 3D4/2 and polarized by cytokines. The 3D4/2 macrophages were treated with 20 ng/mL interferon gamma (IFN-γ) and 10 ng/mL interleukin-4 (IL-4) combined with 10 ng/mL macrophage colony-stimulating factor (M-CSF) to induce polarization to M1 and M2, respectively. After incubation for 24 h, the expression levels of inflammatory factors and iron-metabolism genes were determined using real-time qPCR, Western bot and immunofluorescence. The M1/M2 macrophages culture media supernatant was collected and used to treat porcine intestinal epithelial cells IPEC-J2. The proliferation ability of IPEC-J2 was detected using CCK-8 assay kit. Following exogenous addition of ammonium ferric citrate (FAC) to M1/M2 macrophages, the phagocytic function of macrophages was detected using fluorescein isothiocyanate-dextran (FITC-dextran) and flow cytometry. The results showed that, compared with control, M1 macrophages had higher mRNA levels of iron storage proteins (ferritin heavy and light polypeptide, i.e. FtH and FtL), hepcidin and lipocalin-2, as well as iron content. Moreover, iron enhanced the ability of M1 macrophages to phagocytize FITC-dextran. There was no significant change in these mRNA expression levels in M2 macrophages, but the mRNA expression levels of ferroportin and transferrin receptor were up-regulated. In addition, the conditioned media supernatant from M2 macrophages promoted cell proliferation of IPEC-J2. These findings indicate that M1 macrophages tend to lock iron in the cell and reduce extracellular iron content, thereby inhibiting the proliferation of extracellular bacteria. While M2 macrophages tend to excrete iron, which contributes to the proliferation of surrounding cells and thus promotes tissue repair.

Improved quantification of immune-gold labeling and its use to compare the distribution of cellular factors among sub-chloroplast compartments.

Quantification of immuno-gold labeling can provide valuable information on the quantity and localization of a target within a region of interest (ROI). Background subtraction usually requires preparation of material with a deliberately reduced amount of target component often by gene knockout/knockdown. This paper reports a modified method without the need for gene knockout/knockdown, by using a region outside the ROI as a background and non-immune serum to verify the reliability of the data. An optimized parameter for use in image processing was also developed to improve semi-automatic segmentation of gold particles, by using the standard deviation of pixel intensity together with default parameters (size and intensity) to improve specificity. The modified methods were used to quantify the gold labeling of various components within chloroplasts and their 3 sub-organelle compartments (thylakoid, stroma and starch). Rubisco, actin, myosin, ß-tubulin, Endoplasmic reticulum-retention signal HDEL, Sterol methyltransferase 1, and double stranded RNA were all effectively and consistently quantified at the level of the different sub-chloroplast compartments. The approach should be applicable more widely for high resolution labelling of samples in which a background requiring gene knockout/knockdown is not a realistic option.

Plant begomoviruses subvert ubiquitination to suppress plant defenses against insect vectors.

Most plant viruses are vectored by insects and the interactions of virus-plant-vector have important ecological and evolutionary implications. Insect vectors often perform better on virus-infected plants. This indirect mutualism between plant viruses and insect vectors promotes the spread of virus and has significant agronomical effects. However, few studies have investigated how plant viruses manipulate plant defenses and promote vector performance. Begomoviruses are a prominent group of plant viruses in tropical and sub-tropical agro-ecosystems and are transmitted by whiteflies. Working with the whitefly Bemisia tabaci, begomoviruses and tobacco, we revealed that C2 protein of begomoviruses lacking DNA satellites was responsible for the suppression of plant defenses against whitefly vectors. We found that infection of plants by tomato yellow leaf curl virus (TYLCV), one of the most devastating begomoviruses worldwide, promoted the survival and reproduction of whitefly vectors. TYLCV C2 protein suppressed plant defenses by interacting with plant ubiquitin. This interaction compromised the degradation of JAZ1 protein, thus inhibiting jasmonic acid defense and the expression of MYC2-regulated terpene synthase genes. We further demonstrated that function of C2 protein among begomoviruses not associated with satellites is well conserved and ubiquitination is an evolutionarily conserved target of begomoviruses for the suppression of plant resistance to whitefly vectors. Taken together, these results demonstrate that ubiquitination inhibition by begomovirus C2 protein might be a general mechanism in begomovirus, whitefly and plant interactions.

Instantaneous frequency from Hilbert-Huang transformation of digital volume pulse as indicator of diabetes and arterial stiffness in upper-middle-aged subjects.

To investigate the value of decomposed short-time digital volume pulse (DVP) signals in discerning systemic vascular anomaly in diabetic patients, demographic and anthropometric parameters, serum lipid profile, fasting blood glucose and glycated hemoglobin (HbA1c) levels were obtained from 29 healthy adults (Group 1) and 29 age-matched type 2 diabetes mellitus patients (Group 2). Six-second DVP signals from right index finger acquired through photoplethysmography were decomposed using ensemble empirical mode decomposition. Using one intrinsic mode function (IMF5), stiffness index (SI) and instantaneous energy of maximal energy (fEmax) were obtained. Other indicators of arterial stiffness, including electrocardiogram-pulse wave velocity of foot (ECG-PWVfoot), crest time (CT) and crest time ratio (CTR), were obtained from the testing subjects for comparison. The mean body weight, body mass index, waist circumference, HbA1c and fasting blood sugar levels were higher in Group 2 than those in Group 1, whereas values of systolic and diastolic blood pressure were lower in Group 2 than those in Group 1. SI and fEmax were significantly higher in Group 2 than those in Group 1. Moreover, fEmax was positively associated with HbA1c concentration, CT and SI in Group 2 (p < 0.05) but not in Group 1. When all subjects were considered, fEmax was highly significantly associated with HbA1c and fasting blood sugar levels, and SI (all p < 0.001). After Hilbert-Huang transformation, short-time DVP signals could give significant information on arterial stiffness and vascular anomaly in diabetic patients.

[Catalytic Degradation of Diclofenac Sodium over the Catalyst of 3D Flower-like alpha-FeOOH Synergized with H2O2 Under Visible Light Irradiation].

Three dimensional (3D) flower-like alpha-FeOOH nanomaterials were prepared by oil bath reflux method using FeSO4, urea, ethanol and water, and the products which were characterized by XRD, FT-IR and SEM techniques. The SEM images showed that the 3D flower-like samples consisted of nanorods with a length of 400-500 nm and a diameter of 40-60 nm. The catalytic performance of the samples was evaluated by catalytic degradation of diclofenac sodium using H2O2 as the oxidant under simulated visible light. The results showed that the as-prepared samples presented high efficient catalytic performances, and more than 99% of the initial diclofenac sodium (30 mg x L(-1)) was degraded in 90 min. A radical mechanism can be proposed for the catalytic degradation of diclofenac sodium solution.

Effect of Bak Foong Pills on the expression of ß-amyloid in rat retina with optic nerve transection.

AIM: To investigate the effect of Bak Foong Pills (BFP) on the expression of ß-amyloid (Aß) in rats retina with optic nerve transection, and its roles and possible mechanisms in protecting optic nerve damage. METHODS: Seventy-two healthy, Sprague-Dawley, adult rats were randomly assigned to three groups: negative control group (control group), optic nerve transection group (model group) and BFP treatment group (BFP group, 100µg/mL) followed by establishing optic nerve transection model. The expression of Aß was measured at 48 hours by Western-blotting. Moreover, the expressions of Bcl-2, Bax and Caspase-3 mRNA were evaluated at 48 hours by reverse transcriptase polymerase chain reaction (RT-PCR). RESULTS: There were significant differences among the control, model and BFP groups in the expression of Aß (all P<0.01). Aß expression was significantly higher in the model and BFP groups than that in the control group (P<0.01), with a more significant reduction in the BFP group than that in the model group (P<0.01). Moreover, there were also significant differences among the three groups in the expressions of Bcl-2/Bax (Bcl-2: anti-apoptotic; Bax: proapoptotic) and Caspase-3 mRNA (proapoptotic) (all P<0.01). Bcl-2/Bax ratio was significantly lower and Caspase-3 mRNA expression was significantly higher in the model and BFP groups than those in the control group (P<0.01), with a significant growing of Bcl-2/Bax and reduction of Caspase-3 in the BFP group than those in the model group (P<0.01). CONCLUSION: BFP can down-regulate Aß expression in retina and may inhibit apoptosis and protect optic nerve by enhancing Bcl-2/Bax ratio and inhibiting Caspase-3 pathway.

Co-culture of human extravillous cytotrophoblasts and decidual stromal cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop a convenient method for isolation and purification of human extravillous cytotrophoblasts (EVCTs) and decidual stromal cells (DSCs) and establish a co-culture system.</p><p><b>METHODS</b>The DSCs were digested with trypsin and purified by Percoll gradient. The EVCTs were digested with trypsin and purified by BSA gradient. Immunohischemistry and immunofluorescent study are performed to characterize these isolated cells. The EVCTs and DSCs were placed in Matrigel-coated Transwell upper and lower chamber, respectively, to study the invasive ability of the EVCTs.</p><p><b>RESULTS</b>Immunohischemistry revealed that the purity of EVCTs and DSC exceeded 95%. Cultured EVCTs retained their capacity to invade Matrigel-coated Transwell filters with the invasion index of 3.22-/+0.04.</p><p><b>CONCLUSION</b>This co-culture model established by isolating highly purified EVCTs and DSCs in vitro can be useful for studying the trophoblast invasion mechanisms.</p>

Expression of a begomoviral DNAbeta gene in transgenic Nicotiana plants induced abnormal cell division.

An increasing number of monopartite begomoviruses are being identified that a satellite molecule (DNAbeta) is required to induce typical symptoms in host plants. DNAbeta encodes a single gene (termed betaC1) encoded in the complementary-sense. We have produced transgenic Nicotiana benthamiana and N. tabacum plants expressing the betaC1 gene of a DNAbeta associated with Tomato yellow leaf curl China virus (TYLCCNV), under the control of the Cauliflower mosaic virus 35S promoter. Transgenic plants expressing betaC1 showed severe developmental abnormalities in both species. Microscopic analysis of sections of both transgenic and non-transgenic N. tabacum leaves showed abnormal outgrowths of transgenic N. tabacum to be due to disorganized cell division (hyperplasia) of spongy and palisade parenchyma. Immuno-gold labeling of sections with a polyclonal antibody against the betaC1 protein showed that the betaC1 protein accumulated in the nuclei of cells. The possible biological function of the betaC1 protein was discussed.

[Prokaryotic expression and immuno-localization of replication protein gene of Tobacco curly shoot virus].

The replication protein (Rep) gene of Tobacco curly shoot virus (TbCSV) Y35 isolate was obtained from the infected tobacco plants by PCR and cloned into expression vector pGEX-4T-1 to generate the recombinant plasmid pGEX-Y35Rep. The recombinant plasmid was introduced into Escherichia coli strain BL21(DE3) pLys S, and TbCSV Rep fusion protein was expressed with induction by IPTG and some products were soluble. The Rep fusion protein was purified with GST-Sepharose 4B affinity chromatography and its polyclonal antibody was produced in a rabbit. Studies on subcellular distribution of TbCSV Rep protein in infected tobacco leaves revealed that Rep protein mainly exists in the fractions containing the nucleus. Immuno-gold labeling with the antibody against Rep fusion protein also indicated that Rep protein localized in the nuclei of infected tobacco cells.

Effect of nitric oxide synthase inhibitor on proteoglycan metabolism in repaired articular cartilage in rabbits / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To study the effect of nitric oxide synthase inhibitor, S-methyl thiocarbamate (SMT), on proteoglycan metabolism in repaired articular cartilage in rabbits.</p><p><b>METHODS</b>Twenty-four male New Zealand white rabbits, aged 8 months and weighing 2.5 kg+/-0.2 kg, were used in this study. Cartilage defects in full thickness were created on the intercondylar articular surface of bilateral femurs of all the rabbits. Then the rabbits were randomly divided into 3 groups (n=8 in each group). The defects in one group were filled with fibrin glue impregnated with recombinant human bone morphogenetic protein-2 (rhBMP-2, BMP group), in one group with fibrin glue impregnated with rhBMP-2 and hypodermic injection with SMT (SMT group) and in the other group with nothing (control group). All the animals were killed at one year postoperatively. The tissue sections were stained with safranine O-fast green and analyzed by Quantiment 500 system to determine the content of glycosaminoglycan through measuring the percentage of safranine O-stained area, the thickness of cartilages and the mean gray scale (average stain intensity). Radiolabelled sodium sulphate (Na(2)(35)SO(4)) was used to assess the proteoglycan synthesis.</p><p><b>RESULTS</b>At one year postoperatively, the percentage of safranine O-stained area, the mean gray scale and the cartilage thickness of the repaired tissues in SMT group were significantly higher than those of BMP group (P<0.01) and the control group (P<0.05). Result of incorporation of Na(2)(35)SO(4) showed that the proteoglycan synthesis in SMT group was higher than those of BMP group and the control group (P<0.01).</p><p><b>CONCLUSIONS</b>SMT, a nitric oxide synthase inhibitor, can significantly increase the content of glycosaminoglycan and proteoglycan synthesis, and computer-based image analysis is a reliable method for evaluating proteoglycan metabolism.</p>