Tracking tumor alteration in glioma through serum fibroblast activation protein combined with image.

PURPOSE: Detecting tumor progression of glioma continues to pose a formidable challenge. The role of fibroblast activation protein (FAP) in gliomas has been demonstrated to facilitate tumor progression. Glioma-circulating biomarkers have not yet been used in clinical practice. This study seeks to evaluate the feasibility of glioma detection through the utilization of a serum FAP marker. METHODS: We adopted enzyme-linked immunosorbent assay (ELISA) technique to quantify the relative FAP level of serum autoantibodies in a cohort of 87 gliomas. The correlation between preoperative serum autoantibody relative FAP levels and postoperative pathology, including molecular pathology was investigated. A series of FAP tests were conducted on 33 cases of malignant gliomas in order to ascertain their efficacy in monitoring the progression of the disease in relation to imaging observations. To validate the presence of FAP expression in tumors, immunohistochemistry was conducted on four gliomas employing a FAP-specific antibody. Additionally, the investigation encompassed the correlation between postoperative tumor burden, as assessed through volumetric analysis, and the relative FAP level of serum autoantibodies. RESULTS: A considerable proportion of gliomas exhibited a significantly increased level of serum autoantibody relative FAP level. This elevation was closely associated with both histopathology and molecular pathology, and demonstrated longitudinal fluctuations and variations corresponding to the progression of the disease The correlation between the rise in serum autoantibody relative FAP level and tumor progression and/or exacerbation of symptoms was observed. CONCLUSIONS: The measurement of serum autoantibody relative FAP level can be used to detect the disease as a valuable biomarker. The combined utilization of its detection alongside MR imaging has the potential to facilitate a more accurate and prompt diagnosis.

The extract of Gnaphalium affine D. Don protects against H2O2-induced apoptosis by targeting PI3K/AKT/GSK-3ß signaling pathway in cardiomyocytes.

ETHNOPHARMACOLOGICAL RELEVANCE: Gnaphalium affine D. Don is an important Traditional Chinese herbal Medicine (TCM) used to treat hyperuricemia, asthma, rheumatic arthritis, antitussive, expectorant and cardiovascular in folk medicine because of anti-inflammatory and anti-oxidant activity. The aim of this study was to investigate the potential beneficial effect of G. affine extract (GAE) on hydrogen peroxide (H2O2)-induced apoptosis and explore the possible underlying mechanism in cardiomyocyte. MATERIALS AND METHODS: The ingredients of GAE were isolated and tentatively identified by HPLC-ESI-Q-Qribatrip-MS/MS. The cardioprotective and anti-oxidant effects of GAE were evaluated in the experimental model with H2O2 induced apoptosis in H9c2 cells. H9c2 cells were pretreated for 3 h with or without GAE or with GAE plus PX866 (PI3K inhibitor), then exposed to H2O2 for 6 h, H9c2 cells viability were detected by CCK8 kit, the content of intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) and intracellular superoxide dismutase (SOD) activity were measured by the commercial biochemical kits, western blotting, immunohistochemical (IHC), immunofluorescence (IF) and reverse transcription-polymerase chain reaction (RT-PCR) assays were performed to evaluate the proteins and mRNA expression, propidium iodide (PI) staining was adopted to indicate H9c2 cells apoptosis. RESULTS: Firstly, seventeen polyphenols and flavonoids compounds with the characteristics of anti-inflammatory and anti-oxidant in GAE were tentatively identified by HPLC-ESI-Q-Qribatrip-MS/MS. In the experimental model, GAE not only significantly improved cells viability, but also showed anti-oxidant effects through improving SOD activity, up-regulating nuclear factor E2-related factor 2 (Nrf2), and decreasing intracellular concentration of ROS and MDA and the proteins expression of p47phox, p67phox and gp91phox. On the other hand, GAE revealed anti-apoptotic effect through up-regulating the expression of B-cell lymphoma-2 (Bcl-2), down-regulating Bcl2-associated X (BAX) and cleaved-caspase 3. Furthermore, GAE significantly facilitated phosphorylation of AKT and glycogen synthase kinase-3 beta (GSK-3ß) but not AMPK, while the effects were blocked by PX866 (PI3K inhibitor). CONCLUSIONS: Our data suggested that GAE showed strong anti-oxidant effect to ameliorate oxidative stress and attenuate apoptosis induced by H2O2 in H9c2 cells by targeting PI3K/AKT/GSK-3ß signaling pathway.

Usefulness of a combined approach of DIERS Formetric 4D® and QUINTIC gait analysis system to evaluate the clinical effects of different spinal diseases on spinal-pelvic-lower limb motor function.

BACKGROUND: To investigate the alterations in body movement and their compensatory characteristics under different spinal diseases through an objective and quantitative analysis of the spinal-pelvic-lower limb motor function. METHODS: A total of 120 subjects were recruited from October 2016 to April 2017. The patients were classified into 2 groups in which 65 patients with cervical spondylotic myelopathy (CSM) and 25 patients with idiopathic scoliosis (IS). The former group was evaluated with JOA score while those in the IS group underwent Lenke classification. A control group was set up with 30 healthy subjects. All the subjects were instructed to walk at a constant speed for one minute on a treadmill, and their spinal-pelvic-lower limb motions were monitored simultaneously with a DIERS Formetric 4D® grating system and a QUINTIC gait analysis system. RESULTS: The rotation angle of thoracic and lumbar vertebrae in IS group were larger than those in the control group (P < 0.05), and the knee joint angle A in the CSM group and IS group were larger than the control group (P < 0.05). In the CSM group, the knee joint angular velocity and angular acceleration were both greater than the control group (P < 0.05). And there was a negative linear correlation between the JOA score for the lower extremity of CSM patients and their knee joint angular acceleration. CONCLUSION: IS patients tend to demonstrate increased swing amplitude of the trunk. Those with CSM will also have larger knee joint angular velocity and angular acceleration.

Horseshoes in 4-dimensional piecewise affine systems with bifocal heteroclinic cycles.

By studying the Poincaré map in a neighborhood of the bifocal heteroclinic cycle (the corresponding subsystems only have conjugate complex eigenvalues), this paper provides a result on the existence of chaotic invariant sets for the two-zone 4-dimensional piecewise affine systems with bifocal heteroclinic cycles that cross the switching manifold transversally at two points. Different from Shil'nikov type theorems, the existence of chaotic invariant sets near the heteroclinic cycles depends not only on the eigenvalue conditions but also on the way of intersections of the stable manifolds and unstable manifolds of the subsystems.

Curcumin protects against diabetic cardiomyopathy by promoting autophagy and alleviating apoptosis.

The effects of curcumin on regulating cardiac apoptosis and autophagy were analyzed in diabetic models both in vivo and in vitro. In vivo, experimental diabetes was induced in mice by low-dose STZ injection combined with a high-fat diet. In vitro, cultured H9c2 cardiomyoblasts were exposed to high d-glucose concentrations combined with palmitate. Our results showed that apoptosis was increased and autophagy was suppressed in the hearts of diabetic mice, which was ameliorated by curcumin treatment, ultimately improving cardiac function. Moreover, the inhibition of autophagy exacerbated apoptotic death in cardiac cells under diabetic condition. Curcumin activated AMPK and JNK1, which phosphorylated Bcl-2 and Bim and subsequently disrupted their interactions with Beclin1, thereby promoting autophagy and alleviating apoptosis respectively. In addition, AMPK-mediated inhibition of mTORC1 pathway likely played a role in regulating autophagy by curcumin under diabetic condition. Our study suggests that curcumin protects against diabetic cardiomyopathy by modulating the crosstalk between autophagic and apoptotic machinery. Modulation of autophagy may be an effective strategy for the treatment of cardiovascular diseases associated with diabetes.

Evaluation of the antioxidant and endothelial protective effects of Lysimachia christinae Hance (Jin Qian Cao) extract fractions.

BACKGROUND: Lysimachia christinae Hance is a traditional Chinese medicine with diuretic, detumescent, and detoxifying effects. Our aimed to optimize the extraction protocol to maximize the yield of flavonoids from Lysimachia christinae Hance, and evaluate the pharmacological activities of four fractions, namely, petroleum ether (PE), ethyl acetate (EA), n-butanol (NB), and aqueous (AQ) fractions, of the ethanolic extract of Lysimachia christinae Hance. METHODS: The flavonoid monomers in the crude extract were characterized via high performance liquid chromatography (HPLC), were used as markers for extract quality control and standardization. The total flavonoid, total phenolic, and total polysaccharide contents of each fraction were determined by spectrophotometry. Further, the in vitro free radical (diphenylpicrylhydrazyl, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), superoxide, and hydroxyl radicals) scavenging activities, and antioxidant capacity in endothelial cells were evaluated for each fraction. RESULTS: After optimizing the extraction protocol to maximize the total flavonoid yield from L. christinae Hance, the NB fractions had the highest total flavonoid (39.4 ± 4.55 mg RE/g), total phenolic (41.1 ± 3.07 mg GAE/g) and total polysaccharide (168.1 ± 7.07 mg GE/g); In addition, the NB fraction of the ethanolic extract of L. christinae Hance reveal the strongest radical-scavenging activity, antioxidant activity and protective effects against H2O2-induced injury in HUVECs. CONCLUSIONS: Among the four fractions of L. christinae Hance, the NB fraction showed the most potent antioxidant and endothelial protective effects, which may be attributed to its high flavonoid, phenolic contents and optimal portfolio of different active ingredients of NB fractions of the ethanolic extract of L. christinae Hance. This study might improve our understanding of the pharmacological activities of L. christinae Hance, thereby facilitating its use in disease prevention and treatment.

Curcumin administration suppresses collagen synthesis in the hearts of rats with experimental diabetes.

Cardiac fibrosis is considered the initial change of diabetic cardiomyopathy (DCM). We have shown that curcumin alleviates collagen deposition in DCM, but the mechanism remains unknown. In this study we sought to investigate the effects of curcumin on cardiac fibrosis in vivo and in vitro and to elucidate the underlying mechanisms. Experimental diabetes was induced in rats by injection of low-dose streptozotocin (STZ) combined with high energy diet. The rats were orally treated with curcumin (300 mg·kg-1·d-1) for 16 weeks. Curcumin administration significantly suppressed the deposition of type I and type III collagens in the heart tissues of diabetic rats, accompanied by markedly reduced TGF-ß1 production, suppressed TßR II levels and Smad2/3 phosphorylation, and increased Smad7 expression. Similar effects were observed in human cardiac fibroblasts exposed to high glucose (HG, 30 mmol/L) or exogenous TGF-ß1 (5 ng/mL). Furthermore, TGF-ß1 or HG treatment significantly increased the phosphorylation levels of AMPK and p38 MAPK in the fibroblasts. Application of curcumin (25 µmol/L) inhibited TGF-ß1- or HG-induced AMPK/p38 MAPK activation and suppressed collagen synthesis in the fibroblasts. These effects were similar to those of the AMPK inhibitor compound C (10 µmol/L) but opposite to the effects of the AMPK activator metformin (2 mmol/L) in the fibroblasts. Our results demonstrate that curcumin suppresses diabetes-associated collagen synthesis in rat myocardium not only by inhibiting TGF-ß1 production and canonical Smad signaling but also by blocking the non-canonical AMPK/p38 MAPK pathway.

Dynamic desorption of arsenic from polymer-supported hydrated iron(III) oxide in a wastewater treatment plant.

Polymer-supported hydrated iron(III) oxide (PHIO) was successfully applied as adsorbent for arsenic removal in a wastewater treatment plant in Nandan, China. The practical PHIO adsorbent samples (PHIO-P) were collected from the adsorption column of the wastewater treatment plant, and desorption experiments of the adsorbent were carried out. Our results showed that the formation of precipitates on the surface of PHIO-P might block the porous channel of the adsorbent and decrease its arsenic adsorption capacity. In the dynamic arsenic desorption experiment, the arsenic desorption equilibrium was achieved more quickly at decreasing desorption velocity, and higher arsenic desorption efficiency was obtained at increasing NaOH concentration in regenerant. It was found that the PHIO-P adsorbent could be well regenerated at 1.0 M NaOH solution and desorption velocity of 5 BV h-1. Comparing with the raw adsorbent, the maximum arsenic adsorption capacity of PHIO-P decreased by 41.1% after practical running for 26 months. Additionally, the frequently used waste PHIO adsorbent could be treated as non-hazardous material in the arsenic-containing wastewater treatment process after long-time use.

[Technology and MethodsSimultaneous Determination of 9 Natural Ingredientsin Functional Food for Anti-hangover and Hepatoprotection by High Performance Liquid Chromatography].

OBJECTIVES: To develop a rapid method of high performance liquid chromatography coupled with variable wave length UV detection for simultaneous determination of 9 natural functional ingredients including puerarin, silymarin, quercetin hydrate, schisandrol A, curcumin, tanshinoneI, tanshinoneIIA, cryptotanshinone, and dihydrotanshinoneIin functional food for anti-hangover and hepatoprotection. METHODS: The samples were ultrasonically extracted with 90 % ethanol (V/V) and centrifuged at 10 000 r/min for 10 min prior to HPLC analysis. The nine target analytes were separated on a C18 column with gradient elution using methanol and water (The pH value was adjusted to 2.5 using H3PO4) as the mobile phase. Qualitative analysis was carried out using retention times of the chromatographic peaks, while the external standard curves were established for quantification. RESULTS: Under the optimal analytical conditions, the peak area of each analyte and its concentration had a good correlation within the linear range ( r≥0.998). The limits of detection and quantification of the method were 0.38-0.73 mg/kg (S/N=3) and 1.27-2.43 mg/kg (S/N=10), respectively. The spiked recoveries of the analytes were 88.9%-103.2%, and the relative standard deviations were 1.3%-3.7%. CONCLUSIONS: The method for the determination of 9 functional ingredients in functional food for anti-hangover and hepatoprotection was proposed for the first time in this study. The results showed that it could meet the requirement of routine analysis and quality control and evaluation.

Adsorption characteristic of As(III) on goethite waste generated from hydrometallurgy of zinc.

In this paper, goethite waste from hydrometallurgy of zinc was used as a raw material for arsenic adsorbent preparation. The goethite waste adsorbent (GWA) was characterized with scanning electron microscope (SEM), X-ray powder diffraction (XRD), and particle size distribution analysis. The adsorption of As(III) on GWA was studied as a function of contact time, pH, and coexisting anions. The safety of GWA usage in the wastewater treatment process was assessed by toxicity characteristic leaching procedure (TCLP) tests. The equilibrium adsorption data fitted well with the Langmuir isotherm model, and the maximum adsorption capacity of As(III) on GWA was 51.47 mg.g-1. GWA showed higher adsorption capacity at weak alkaline pH values (7.0-9.5). The coexisting PO43- and SiO32- presented significant adsorption competition with As(III) in aquatic systems. No significant heavy metals leaching was observed for GWA and As(III) loaded GWA in TCLP tests, which implied the safety of GWA as an adsorbent for arsenic containing wastewater treatment.

Topological horseshoe and its uniform hyperbolicity in the HP model.

A famous food-chain model proposed by Hastings and Powell is numerically restudied. The existence and uniform hyperbolicity of chaotic invariant sets are demonstrated by means of the topological horseshoe theory and the Conley-Moser conditions, indicating that, for a fixed cross section, the second return Poincaré map of the model possesses a closed uniformly hyperbolic chaotic invariant set, on which it is topologically conjugate to the 2-shift map.

On the existence of bifocal heteroclinic cycles in a class of four-dimensional piecewise affine systems.

Based on mathematical analysis, this paper provides a methodology to ensure the existence of heteroclinic cycles in a class of four-dimensional piecewise affine systems. In addition, examples are provided to illustrate the effectiveness of the method.

A vast amount of various invariant tori in the Nosé-Hoover oscillator.

This letter restudies the Nosé-Hoover oscillator. Some new averagely conservative regions are found, each of which is filled with different sequences of nested tori with various knot types. Especially, the dynamical behaviors near the border of "chaotic region" and conservative regions are studied showing that there exist more complicated and thinner invariant tori around the boundaries of conservative regions bounded by tori. Our results suggest an infinite number of island chains in a "chaotic sea" for the Nosé-Hoover oscillator.

Bifurcation and chaos in the simple passive dynamic walking model with upper body.

We present some rich new complex gaits in the simple walking model with upper body by Wisse et al. in [Robotica 22, 681 (2004)]. We first show that the stable gait found by Wisse et al. may become chaotic via period-doubling bifurcations. Such period-doubling routes to chaos exist for all parameters, such as foot mass, upper body mass, body length, hip spring stiffness, and slope angle. Then, we report three new gaits with period 3, 4, and 6; for each gait, there is also a period-doubling route to chaos. Finally, we show a practical method for finding a topological horseshoe in 3D Poincaré map, and present a rigorous verification of chaos from these gaits.

Secretory expression of a bispecific antibody targeting tumor necrosis factor and ED-B fibronectin in Pichia pastoris and its functional analysis.

Specific targeting of tumor necrosis factor (TNF)-α antagonist to the inflamed site could increase its efficacy and reduce side-effects. Here, we constructed a bispecific diabody (BsDb) that targets TNF-α and ED-B-containing fibronectin, a fibronectin isoform specifically expressed in the pannus of the inflamed synovium in rheumatoid arthritis. BsDb was secreted from Pichia pastoris as functional protein and was purified to homogeneity. BsDb could simultaneously bind to human TNF-α and B-FN and neutralize TNF-α action. Additionally, BsDb showed a significant gain both in the antigen-binding affinity and in TNF-α-neutralizing ability as compared to its original antibodies, L19 and anti-TNF-α scFv, which were produced in E. coli. BsDb was constructed and was endowed with enhanced bioactivities and improved production processing. Therefore, it holds great potential for in vivo applications.

Protein kinase C-δ mediates sepsis-induced activation of complement 5a and urokinase-type plasminogen activator signaling in macrophages.

OBJECTIVE AND DESIGN: Activations of the complement C5a (C5a) and the urokinase-type plasminogen activator (uPA) are commonly seen together during sepsis. However, the mechanism linking these two important pathways remains elusive. MATERIAL, METHODS AND TREATMENT: We used the C57BL/6 J mice model of sepsis induced by cecal ligation puncture (CLP) procedure, injected anti-C5aR or rottlerin through the tail vein to neutralize C5aR or PKC-δ, and then isolated peritoneal macrophages. Total RNA was isolated from the cells and analyzed by quantitative PCR. RESULTS: Our study revealed that neutralizing C5aR markedly inhibited sepsis-induced uPA receptor (uPAR) expression and its downstream signaling in macrophage. Similarly, neutralizing uPAR suppressed sepsis activation of C5a signaling. Importantly, inhibition of PKC-δ largely blocked sepsis-induced expression of C5aR and uPAR. CONCLUSIONS: Our study demonstrates a crosstalk between the complement C5a signaling and the fibrinolytic uPA pathways, which may depend on each other to maintain their expression and signaling, and reveals a central role of PKC-δ in mediating sepsis-induced activation of these pathways.

64-Slice spiral CT perfusion combined with vascular imaging of acute ischemic stroke for assessment of infarct core and penumbra.

The aim of this study was to determine the value of computed tomography perfusion (CTP) parameters, including cerebral blood flow (CBF), cerebral blood volume (CBV), mean transit time (MTT) and time-to-peak (TP), in a clinical study of patients with stroke. Additionally, we determined which parameter or combination of parameters are reliable in detecting the presence of an infarct and penumbra. CTP was performed within 24 h of the onset of symptoms in 20 patients with possible stroke. Magnetic resonance imaging (MRI) was performed 3-7 days later and the threshold of the CTP was adjusted according to the results to provide CT images that correlated with the MRI; the MRI results were taken as the gold standard. CBV, CBF and TP contrast agent enhancement were calculated using the CT results. The CTP results were compared with the MRI findings. All CTP parameters were reliable in detecting the penumbra (P<0.001). In these parameters, changes of MTT were the most useful. CTP revealed various changes in CBF, CBV, MTT and TP in ischemic areas. CTP parameters were also reliable in detecting the infarct core (P<0.001). We determined that when detecting the penumbra, all CTP parameters are reliable, and when detecting cerebral ischemia, a combination of parameters should be used.

New bifurcations in the simplest passive walking model.

This paper uncovers several new stable periodic gaits in the simplest passive walking bipedal model proposed in the literature. It is demonstrated that the model has period-3 to period-7 gaits beside the period-1 gaits found by Garcia et al. By simulations, this paper shows that each of these new gaits leads to chaos via period-doubling bifurcation and loses its stability by cyclic-fold bifurcation. This interesting phenomenon suggests a series of new bifurcation scenarios that have not been observed before. To confirm the new gaits and their bifurcations, this paper presents computer assisted proofs on the existence and stability of each periodic gait and its period-doubling gaits with the interval Newton method. To verify that the routes indeed lead to chaos, computer-assisted proofs are also given by means of topological horseshoes theory.

Cell-surface display of the active mannanase in Yarrowia lipolytica with a novel surface-display system.

A novel surface-display system was constructed using the cell-wall anchor protein Flo1p from Saccharomyces cerevisiae, the mannanase (man1) from Bacillus subtilis fused with the C-terminus of Flo1p and the 6xHis tag was inserted between Flo1p and man1. The fusion protein was displayed on the cell surface of Yarrowia lipolytica successfully, and it was confirmed by immunofluorescence. In succession, the surface-displayed mannanase was characterized. The optimum catalytic conditions for the recombinant mannanase were 55 degrees C at pH 6.0, and it exhibited high stability against pH variation. The highest activity of the recombinant mannanase reached 62.3 IU/g (dry cell weight) after the recombinant was cultivated for 96 h in YPD medium [1% (w/v) yeast extract/2% (w/v) peptone/2% (w/v) glucose]. To our knowledge, the present paper is the first to report that high-activity mannanase is displayed on the cell surface of Y. lipolytica with Flo1p.