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Flower-like ZnO microstructures were successfully produced using a hydrothermal method employing ZnSO4/(NH4)2SO4 as a raw material. The effect of the operating parameters of the hydrothermal temperature, OH−/Zn2+ molar ratio, time, and amount of dispersant on the phase structure and micromorphology of the ZnO particles were investigated. The synthesis conditions of the flower-like ZnO microstructures were: hydrothermal temperature of 160°C, OH−/Zn2+ molar ratio of 5:1, reaction time of 4 h, and 4 mL of dispersant. The flower-like ZnO microstructures were comprised of hexagon-shaped ZnO rods arranged in a radiatively. Degradation experiments of Rhodamine B with the flower-like ZnO microstructures demonstrated a degradation efficiency of 97.6% after 4 h of exposure to sunshine, indicating excellent photocatalytic capacity. The growth mechanism of the flower-like ZnO microstructures was presented.

An improved method of (NH4)2SO4 roasting followed by water leaching to utilize zinc oxidized ores was studied. The operating parameters were obtained by investigating the effects of the molar ratio of (NH4)2SO4 to zinc, roasting temperature, and holding time on zinc extraction. The roasting process followed the chemical reaction control mechanism with the apparent activation energy value of 41.74 kJ·mol−1. The transformation of mineral phases in roasting was identified by X-ray diffraction analysis combined with thermogravimetry-differential thermal analysis curves. The water leaching conditions, including the leaching temperature, leaching time, stirring velocity, and liquid-to-solid ratio, were discussed, and the leaching kinetics was studied. The reaction rate was obtained under outer diffusion without product layer control; the values of the apparent activation energy for two stages were 4.12 and 8.19 kJ·mol−1. The maximum zinc extraction ratio reached 96% while the efficiency of iron extraction was approximately 32% under appropriate conditions. This work offers an effective method for the comprehensive use of zinc oxidized ores.

NH4HCO3 conversion followed by HCl leaching was performed and proven to be effective in extracting Pb and Sr from zinc extracted residual. The mechanism and operating conditions of NH4HCO3 conversion, including molar ratio of NH4HCO3 to zinc extracted residual, NH4HCO3 concentration, conversion temperature, conversion time, and stirring velocity, were discussed, and operating conditions were optimized by the orthogonal test. Experimental results indicate that NH4HCO3 conversion at temperatures ranging from 25 to 85°C follows the shrinking unreacted core model and is controlled by inner diffusion through the product layer. The extraction ratios of Pb and Sr under optimized conditions reached 85.15% and 87.08%, respectively. Moreover, the apparent activation energies of Pb and Sr were 13.85 and 13.67 kJ·mol−1, respectively.

Employing zinc sulfate solution obtained from zinc oxide ore as raw material, sodium hydroxide as precipitant and PEG20000 as dispersant, ultrafine ZnO powders with different morphologies were successfully synthesized through hydrothermal method. The influences of the dosage of PEG20000 solution, molar ratio of OH−/Zn2+, reaction temperature, reaction time and Zn2+ concentration on the structures and morphologies of the ZnO powders were discussed in detail. The reaction conditions of synthesizing ZnO powders with flower-like structure were obtained as below: dosage of PEG20000 with 10% mass fraction 5 mL, molar ratio of OH− to Zn2+ 5, reaction temperature 150 °C, reaction time 8 h at Zn2+ concentration 1 mol L−1. The growth mechanism of ZnO particles with different morphologies was proposed. The ZnO powder with flower-like structure are composed of multiple micro-rods with hexagon morphology and has good photocatalytic degradation ability to degrade RhB. 20 mL RhB solution with 15 mg L−1 could be completely degraded over flower-like ZnO powder 300 mg within 3 h.

Although electrospinning of nanofibrous scaffolds benefit to wound healing have been developed at a fast pace, achieving nanofibrous scaffolds with high absorptivity by green electrospinning and crosslinking techniques is still a great challenge. Here, we developed a maleilated hyaluronate/methacrylated poly (vinyl alcohol) (MHA) (MaPVA) composite nanofibers by electrospinning from pure water solvent and followed by photopolymerization to form crosslinking nanofibers network. Electrospinnability of MHA/MaPVA blend systems were investigated and the results shows that the morphology and diameter of the nanofibers were mainly affected by MHA/MaPVA weight ratios by changing viscosity and conductivity of the blend solutions. The crystalline microstructure of the electrospun fibers was not well developed due to intermolecular hydrogen bonding interaction between the molecules of MHA and MaPVA. The photocrosslinking MHA/MaPVA nanofibrous mats can swell to form fibrous hydrogels with high water absorption, meanwhile it is cytocompatible and capable of promoting the cell attachment, which render it great potential for wound dressings.

A major challenge in rice (Oryza sativa L.) production is to cope with increasing grain yield and fertilizer use efficiency without compromising grain quality. This study was designed to determine if optimizing integrative cultivation management in rice could improve grain quality while increase yield and nitrogen use efficiency (NUE). An indica-japonica hybrid rice cultivar and a japonica rice cultivar were grown in the field, with five cultivation managements including no N application (0 N), local farmer's practice (LFP), and three optimizing integrative cultivation managements, reducing N rate and increasing plant density (ND), ND+alternate wetting and moderate soil drying irrigation (NDW), and NDW+applying rapeseed cake fertilizer (NDWR). The results showed that the optimizing integrative cultivation managements could not only increase grain yield, but also enhance NUE compared to LFP. Compared to LFP, NDWR significantly increased brown, milled, head milled rice rate, ratio of the kernel length to breadth and breakdown value of starch, whereas decreased amylose content, gel consistency, prolamin content, setback value, percentage of chalky kernels, and chalkiness. The three optimizing integrative cultivation managements increased contents of total proteins, albumin and glutelin, activities of the key enzymes involved in the sucrose-starch conversion in grains, root oxidation activity, and malic and succinic acid concentrations in root exudates during the grain-filling period. The results suggested that optimizing integrative cultivation managements could improve grain quality meanwhile increase grain yield and NUE by enhancing physiological activities of rice plants.

ZnO@SiO2 composite with flower-like structure was successfully prepared with molar ratio of ZnO/SiO2 = 1:1 based on the optimized synthesizing parameters of spherical SiO2 and flower-like ZnO. SiO2 particles were coated on the flower-like ZnO to form a homogeneous film through the multidimensional polycondensation of Si(OH)4. The photocatalytic degradation ability of ZnO@SiO2 composite for rhodamine B (RhB) obtained at different ZnO/SiO2 molar ratio and the comparison to that of flower-like ZnO showed that ZnO@SiO2 composite with ZnO/SiO2 molar ratio of 1:1 displayed a relatively good photocatalytic degradation ability to degrade RhB, but it was weaker than that of flower-like ZnO. Twenty millilitres of RhB solution at a concentration of 15 mg·L−1 could be completely degraded by 300 mg flower-like ZnO powder within 3 h, while the degrading efficiency was only 82.5% by 300 mg ZnO@SiO2 composite. But ZnO@SiO2 composite showed a better photocatalytic activity than flower-like ZnO at a lower pH value of 4.5.

Crocin is the main monomer of saffron, which is a momentous component of traditional Chinese medicine Lang Qing A Ta. Here, we tried to probe into the role of crocin in liver fibrosis. Hematoxylin-eosin staining and Sirius Red staining were used to observe the pathological changes of liver tissues. After hepatic stellate cells (HSCs) were isolated from liver tissues, lnc-LFAR1, MTF-1, GDNF, and α-SMA expressions were detected by qRT-PCR and western blot. Immunohistochemistry and immunofluorescence were used to detect α-SMA expression. Chromatin immunoprecipitation was used to analyze the binding of MTF-1 to the GDNF promoter. Moreover, the dual-luciferase reporter gene, RNA pull-down, and RNA immunoprecipitation were used to clarify the interaction between MTF-1 and GDNF, lnc-LFAR1 and MTF-1. The degree of liver fibrosis was more severe in the mice from the liver fibrosis model, while the liver fibrosis was alleviated by the injection of crocin. lnc-LFAR1, GDNF, and α-SMA were up-regulated, and MTF-1 was down-regulated in liver fibrosis tissues and cells, while these trends were reversed after the injection of crocin. Besides, lnc-LFAR1 negatively regulated MTF-1 expression, and positively regulated GDNF and α-SMA expressions, and MTF-1 was enriched in the promoter region of GDNF. Furthermore, the cellular direct interactions between MTF-1 and GDNF, lnc-LFAR1 and MTF-1 were verified. In vivo experiments confirmed the relief of crocin on liver fibrosis. Our research expounded that crocin restrained the activation of HSCs through the lnc-LFAR1/MTF-1/GDNF axis, thereby ameliorating liver fibrosis.

Decrease in the grain yield resulted from a low percentage of opened spikelets under high temperature (HT) during anthesis is a serious problem in the seed production of photo-thermo-sensitive genic male sterile (PTGMS) rice (Oryza sativa L.) lines, and the mechanism is little understood. Elucidating the physiological mechanism underlying the effect of HT during anthesis on spikelet-opening of PTGMS lines would have great significance in exploring the effective way to mitigate the adverse effect of HT. In this study, two PTGMS lines and one restorer line of rice were used and were subjected to normal temperature (NT) and HT treatments. The results showed that, compared with NT, HT significantly decreased the percentage of opened spikelets, fertilization percentage and seed-setting by significantly increasing the percentage of wrapped spikelets and reducing the spikelet-opening angle, length of spikelet-opening time. The HT significantly decreased the contents of soluble sugars, jasmonic acid (JA) and methyl jasmonate (MeJA) in the lodicules before and at glume-opening, which were significantly correlated with and accounts for the low percentage of opened spikelets under HT for rice, especially for the PTGMS lines.

The reaction kinetics of roasting zinc silicate using NaOH was investigated. The orthogonal test was employed to optimize the reaction conditions and the optimized reaction conditions were as follows: molar ratio of NaOH to Zn2SiO4 of 16:1, reaction temperature of 550 °C, and reaction time of 2.5 h. In order to ascertain the phases transformation and reaction processes of zinc oxide and silica, the XRD phase analysis was used to analyze the phases of these specimens roasted at different temperatures. The final phases of the specimen roasted at 600 °C were Na2ZnO2, Na4SiO4, Na2ZnSiO4 and NaOH. The reaction kinetic equation of roasting was determined by the shrinking unreacted core model. Aiming to investigate the reaction mechanism, two control models of reaction rate were applied: chemical reaction at the particle surface and diffusion through the product layer. The results indicated that the diffusion through the product layer model described the reaction process well. The apparent activation energy of the roasting was 19.77 kJ/mol.

The orthogonal test was used to optimize the reaction conditions of roasting zinc oxide ore with NaOH aiming to comprehensively utilize zinc oxide ore. The optimized reaction conditions were molar ratio of NaOH to zinc oxide ore 6:1, roasting temperature 450 °C, holding time 150 min. The molar ratio of NaOH to zinc oxide ore was the most predominant factor affecting the extraction ratios of zinc oxide and silica. The mineral phase transformations were investigated by testing the phases of specimens obtained at different temperatures. The process was that silica reacted with molten NaOH to form Na2SiO3 at first, then transformed into Na4SiO4 with temperature rising. ZnCO3 and its decomposing product ZnO reacted with NaOH to form Na2ZnO2. Na2ZnSiO4 was also obtained. The reaction rate was investigated using unreacted shrinking core model. Two models used were chemical reaction at the particle surface and diffusion through the product layer. The results indicated that the reaction rate was combine-controlled by two models. The activation energy and frequency factor were obtained as 24.12 kJ/mol and 0.0682, respectively.

A nanocarrier system of methoxypolyethylene glycol amine (mPEG-NH2) functionalized polydopamine (PDA) coated hollow mesoporous silica nanoparticles (HMSNs-PDA-PEG) was developed with pH-responsive, which combined doxorubicin hydrochloride (DOX) and quercetin (QUR) to reverse multidrug resistance (MDR) and improved anticancer effects on taxol (TAX) and DOX double resistant human colorectal cancer cell line HCT-8 (HCT-8/TAX cells). Well-dispersed nanoparticles (HMSNs-PDA-PEG) were prepared with a dimension of around 170 nm. The surface morphology and chemical properties of HMSNs-PDA-PEG were also successfully characterized by transmission electron microscopy (TEM), X-ray photoelectron spectroscopy (XPS), thermal gravimetric analysis (TGA), X-ray diffraction (XRD), Brunauer-Emmett-Teller (BET) method, Fourier transform infrared spectroscopy (FT-IR) and dynamic light scattering (DLS). Drug release experiments results indicated that DOX and QUR (QD) loaded nanoparticles (HMSNs-PDA-PEG@QD) had similar release kinetic profiles of each drug, which all exhibited highly sensitive to pH value due to the surface PDA coating. Additionally, the HCT-8 cells or HCT-8/TAX cells were employed to assess the cellular uptake and cytotoxicity of various drug-free or drug-loaded HMSNs samples. Meanwhile, a series of biological evaluations demonstrated that the HMSNs-PDA-PEG@QD exhibited remarkable ability to overcome MDR compared with free DOX and HMSNs-PDA-PEG@DOX. Taken together, these results revealed that HMSNs-PDA-PEG@QD was suitable as a prospective and efficient drug delivery nanosystem for overcoming multidrug resistance.

An orthogonal test was used to optimize the reaction conditions of roasting zinc oxide ore using (NH4)2SO4. The optimized reaction conditions are defined as an (NH4)2SO4/zinc molar ratio of 1.4:1, a roasting temperature of 440°C, and a thermostatic time of 60 min. The molar ratio of (NH4)2SO4/zinc is the most predominant factor and the roasting temperature is the second significant factor that governs the zinc extraction. Thermogravimetric–differential thermal analysis was used for (NH4)2SO4 and zinc mixed in a molar ratio of 1.4:1 at the heating rates of 5, 10, 15, and 20 K·min−1. Two strong endothermic peaks indicate that the complex chemical reactions occur at approximately 290°C and 400°C. XRD analysis was employed to examine the transformations of mineral phases during roasting process. Kinetic parameters, including reaction apparent activation energy, reaction order, and frequency factor, were calculated by the Doyle–Ozawa and Kissinger methods. Corresponding to the two endothermic peaks, the kinetic equations were obtained.

A highly sensitive and selective 3D excitation-emission fluorescence method has been proposed to rapidly quantify the combined antidiabetics Repaglinide (Re) and Irbesartan (Ir) in rat and human plasmas with the aid of second-order calibration method based on alternating trilinear decomposition (ATLD) method. Re and Ir with weak fluorescence can be endowed with strong fluorescent property by changing the microenvironment in samples and improving the fluorescence quantum yield by using an appropriate micellar enhanced surfactant. The enhanced excitation-emission matrix fluorescence of Re and Ir can be accurately resolved and can simultaneously attain the optimal concentration even in the presence of a potentially strong intrinsic fluorescence from complex biological matrices, such as rat and human plasmas, by using the ATLD method, which completely exploits the “second-order advantage”. The average recoveries of Re and Ir obtained from ATLD with the factor number of 3 (N=3) were 101.0%±4.3% and 99.1%±4.1% for rat plasma and 100.5%±5.4% and 97.1%±3.6% for human plasma. Several statistical methods, including Student’s t-test, figures of merit, and elliptical joint confidence region, have been utilized to evaluate the accuracy of the proposed method. Results show that the developed method can maintain second-order advantage in simultaneous determinations of the weak fluorescent analytes of interest in different biological plasma matrices.

(NH4)2SO4 roasting followed by water leaching to process nickel- chromium mixed metal oxidized (MO) ore was investigated. The optimal operating parameters were obtained after determining the influences of the molar ratio of (NH4)2SO4 to mixed oxidized ore, roasting temperature and roasting time on metal extractions. The extraction efficiencies of Ni, Fe, Mg, Al and Cr reached 94.02%, 87.87%, 84.14%, 74.00% and 41.49%, respectively. The transformation of mineral phases during roasting were identified by XRD analysis combining with TG-DTA curves and phases transformation analysis of analytic reagents with (NH4)2SO4. The phase transition process in experimental temperature range can be summarized as: MO to (NH4)2M(SO4)2 to MSO4 and M2O3 to (NH4)3M(SO4)3 to NH4M(SO4)2 to M2(SO4)3 (M is divalent and trivalent metallic element). The work demonstrates an effective process to treat nickel-chromium mixed metal oxidized ores. The reaction mechanism of Fe meets the chemical reaction control mechanism, while the reaction mechanism of Ni, Mg and Al follows the mixed control mechanism of the chemical reaction and diffusion controls.

Background/Aims: Salvia miltiorrhiza (SM) contains four major aqueous active ingredients, which have been isolated, purified and identified as danshensu (DSS), salvianolic acid A (Sal-A), salvianolic acid B (Sal-B) and protocatechuic aldehyde (PAL), totally abbreviated as SABP. Although SM is often used to treat various cardiovascular diseases in traditional Chinese medicine, the efficacy and function of optimal compatibility ratio of SM's active ingredients (SABP) in the prevention and treatment of cardiovascular diseases remain uncertain. This study investigated antihypertensive effect and underlying mechanisms of SABP vs. SM lyophilized powder (SMLP) in spontaneously hypertensive rats (SHR) and to establish the ratio of the optimal compatibility of DSS, Sal-A, Sal-B and PAL in improving cardiovascular functions. Methods: The SHRs were treated with either SABP or SMLP and their systolic blood pressures (SBP) were monitored. The isolated thoracic aorta of SHRs was segregated for immunohistochemistry, Hematoxylin-Eosin stain and mRNA and protein expression of NOX4, TGF-β1, Col-I, ET-1, α-SMA and Smad7. Moreover, the adventitial fibroblasts (AFs) were isolated and cultured from SD rats' aorta and the reactive oxygen species (ROS) production was determined after SABP or SMLP treatment. Results: SABP, but not SMLP, significantly reduced SBP, which were accompanied by the inhibited morphological changes in the thoracic aorta and the reduced mRNA and protein expression of NOX4, TGF-β1, Col-I, ET-1 and α-SMA, but the increased Smad 7 expression in SHRs. Moreover, SABP also resulted in a decreased ROS production in AFs of SD rats. Conclusions: These results indicate that SABP, but not SMLP, treatment potently inhibits hypertension through improvements of vascular remodeling and oxidative stress. The present study provides new evidence that the efficacy and function from optimal compatibility ratio of SM active ingredients is much better than its lyophilized powder, which represents a strategy to develop SM's new beneficial effect in improving cardiovascular functions.

Glycosylation of the conserved asparagine residue in the CH2 domain is the most common posttranslational modification of recombinant monoclonal antibodies. Ideally, a consistent oligosaccharide profile should be maintained from early clinical material to commercial material for the development of recombinant monoclonal therapeutics, though variation in the profile is a typical result of process changes. The risk of oligosaccharide variation posed to further development is required to be thoroughly evaluated based on its impact on antibody structure, stability, efficacy and safety. The variation should be controlled within a range so that there is no detrimental impact on safety and efficacy and thus allowing the use of early phase safety and efficacy data to support project advancement to later phase. This review article focuses on the current scientific understanding of the commonly observed oligosaccharides found in recombinant monoclonal antibodies and their impact on structure, stability and biological functions, which are the basis to evaluate safety and efficacy. It also provides a brief discussion on critical quality attribute (CQA) assessment with regard to oligosaccharides based on the mechanism of action (MOA). © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1173-1181, 2017.

According to GB/T3286.1-2012(The determination of calcium oxide and magnesium oxide content in limestone and dolomite), the mathematical model of magnesium oxide content determination in limestone by atomic absorption spectrometry was established. The various uncertainty factors of different elements for a sample were discussed and compared in the testing process. The confidence interval for the measurement result was (0.74±0.03)%,k=2 in uncertainty evaluation .The results showed that the variability of working curve and accuracy of standard solution volume for working curve were main influence factors of uncertainty. It could effectively reduce the uncertainty from the perspective of the main factors,and improve the quality of analysis.

The ZnO powder was successfully obtained by calcining the basic zinc carbonate, which was synthesized from clinker digestion solution of zinc oxide ore using analytical Na2CO3. The factors of mol ratio of Na2CO3to ZnSO4and reaction temperature were discussed at stirring intensity 400 rpm. Both the obtained basic zinc carbonate and ZnO powders were characterized using XRD, EDS and SEM. The thermal decomposition of basic zinc carbonate was also investigated below 600°C. The results show that the ZnO powder was wurzite hexagonal structure and the well-crystallized ZnO powder was uniformly spherical particles and distributed in a narrow range.

To obtain the appropriate conditions for eliminating Fe3+ from NiSO4 solution, the digestion solution of the clinker was used as raw material, which was obtained from roasting the nickel oxide ore with (NH4)2SO4. The ammonium jarosite was successfully synthesized from the solution with analytic grade NH4HCO3. The effects of reaction temperature, reaction time, end pH value of reaction on the removal rate of iron were investigated, and the effect of the initial concentration of Fe3+ was also discussed. All of those factors had significant effects on the removal rate of Fe3+, among which the reaction temperature was the most prominent. The appropriate reaction conditions were concluded as follows: reaction temperature 95 °C, reaction time 3.5 h, end pH value of reaction 2.5 at initial concentration of Fe3+ 19.36 g/L. The physical aspect of (NH4)2Fe6(SO4)4(OH)12 was cluster figure composed of sheet or prismatic particles with smooth surface.

Inductively coupled plasma atomic emission spectroscopy was applied to the determination of boric anhydride in ascharite ores. Pretreatment method, reagents used for acid decomposition and the interferences from matrix and coexistent elements were discussed in details respectively. The analytical spectral lines were selected while experimental parameters were optimized. The results indicated that after the samples were completely dissolved and collected with hydrochloric acid (1+1) in a sealed container with a reflux condenser by the low-temperature heating method, they could be directly analyzed and determined with ICP-AES at optimized operating parameters. The relative standard deviation of the method is less than 1.4% within the measurement range of 0.1% ~ 40% and the recovery rate is in the range of 98.1% ~ 101.0%. The method was proved satisfactory with wide linear range, low detection limit and also high sensitivity and accuracy.

Unilateral damage to the peripheral vestibular receptors precipitates a debilitating syndrome of oculomotor and balance deficits at rest, which extensively normalize during the first week after the lesion due to vestibular compensation. In vivo studies suggest that GABA(B) receptor activation facilitates recovery. However, the presynaptic or postsynaptic sites of action of GABA(B) receptors in vestibular nuclei neurons after lesions have not been determined. Accordingly, here presynaptic and postsynaptic GABA(B) receptor activity in principal cells of the tangential nucleus, a major avian vestibular nucleus, was investigated using patch-clamp recordings correlated with immunolabeling and confocal imaging of the GABA(B) receptor subunit-2 (GABA(B)R2) in controls and operated chickens shortly after unilateral vestibular ganglionectomy (UVG). Baclofen, a GABA(B) agonist, generated no postsynaptic currents in principal cells in controls, which correlated with weak GABA(B)R2 immunolabeling on principal cell surfaces. However, baclofen decreased miniature excitatory postsynaptic current (mEPSC) and GABAergic miniature inhibitory postsynaptic current (mIPSC) events in principal cells in controls, compensating and uncompensated chickens three days after UVG, indicating the presence of functional GABA(B) receptors on presynaptic terminals. Baclofen decreased GABAergic mIPSC frequency to the greatest extent in principal cells on the intact side of compensating chickens, with concurrent increases in GABA(B)R2 pixel brightness and percentage overlap in synaptotagmin 2-labeled terminals. In uncompensated chickens, baclofen decreased mEPSC frequency to the greatest extent in principal cells on the intact side, with concurrent increases in GABA(B)R2 pixel brightness and percentage overlap in synaptotagmin 1-labeled terminals. Altogether, these results revealed changes in presynaptic GABA(B) receptor function and expression which differed in compensating and uncompensated chickens shortly after UVG. This work supports an important role for GABA(B) autoreceptor-mediated inhibition in vestibular nuclei neurons on the intact side during early stages of vestibular compensation, and a role for GABA(B) heteroreceptor-mediated inhibition of glutamatergic terminals on the intact side in the failure to recover function.

Purpose: To establish if impression cytology combined with histochemical and immunocytochemical staining can be used to assess epithelium of the human upper lid margin. Methods: Following an initial eye examination of 40 healthy subjects (19 soft contact lens wearers and 21 non-contact lens wearers, aged 18–57 years), lid margin staining was assessed with lissamine green using slit lamp biomicroscopy and graded (grade 0–3). Impression cytology of the upper lid margin of both eyes was collected, fixed and stained with periodic acid Schiff (PAS) and haematoxylin for cell morphology analysis (Nelson grade) or for immunocytochemistry (keratinization-related proteins: filaggrin, transglutaminase1 (TGase1) and cytokeratin 1/10). Results: In 57% of all subjects, grade 0 lissamine green staining showed a thin line (the Marx line), just posterior to the meibomian gland ducts. Grade 2 or 3 lissamine green staining was observed in 17% of all subjects. There was no difference between contact lens and non-contact lens wearers for lid margin staining or Nelson grade (p = 0.4, Fisher’s exact test). PAS/haematoxylin staining and immunocytochemistry showed transition in epithelial cell morphology, with marginal conjunctival epithelium, mucocutaneous junction and squamous epithelium, adjacent to meibomian gland ducts. This transition in epithelium was associated with differential expression of keratinization-related proteins (filaggrin, cytokeratin 1/10 and TGase1). Conclusion: Lid margin epithelium can be successfully sampled using impression cytology and further characterized using histochemistry and immunocytochemistry staining techniques. This approach can be applied to assess lid margin changes in conditions such as dry eye and meibomian gland dysfunction.

Vestibular compensation refers to the recovery of function occurring after unilateral vestibular deafferentation, but some patients remain uncompensated. Similarly, more than half of the operated chickens compensate three days after unilateral vestibular ganglionectomy (UVG), but the rest remain uncompensated. This review focuses on the studies performed on the principal cells of the chick tangential nucleus after UVG. The tangential nucleus is a major avian vestibular nucleus whose principal cells are all second-order, vestibular reflex projection neurons participating in the vestibuloocular and vestibulocollic reflexes controlling posture, balance, and eye movements. Using whole-cell patch-clamp approach in brain slice preparations, spontaneous spike firing, ionic conductances, and spontaneous excitatory postsynaptic currents (sEPSCs) are recorded in principal cells from controls and operated chickens three days after UVG. In compensated chickens, the proportion of spontaneous spike firing principal cells and their spike discharge rate are symmetric on the lesion and intact sides, with the rates increased over controls. However, in the uncompensated chickens, the spike discharge rate increases on the lesion side, but not on the intact side, where only silent principal cells are recorded. In all the experimental groups, including controls, silent principal cells are distinguished from spontaneous spiking cells by smaller persistent sodium conductances and higher activation thresholds for the fast sodium channel. In addition, silent principal cells on the intact side of uncompensated chickens have larger dendrotoxin-sensitive potassium conductances, with a higher ratio of immunolabeling for surface/cytoplasmic expression of a dendrotoxin-sensitive, potassium channel subunit, Kv1.1. Finally, in compensated chickens, sEPSC frequency is symmetric bilaterally, but in uncompensated chickens sEPSC frequency increased only on the lesion side, where the expression of Kv1.2 decreased in synaptotagmin-labeled terminal profiles on the principal cell bodies. Altogether, the specific sodium and potassium channels important for the development of spike firing pattern and/or presynaptic glutamate release on vestibular reflex projection neurons may be critically involved in changing postsynaptic neuron excitability after vestibular deafferentation.

Vestibular compensation refers to the behavioral recovery after a unilateral peripheral vestibular lesion. In chickens, posture and balance deficits are present immediately following unilateral vestibular ganglionectomy (UVG). After three days, most operated chickens begin to recover, but severe deficits persist in others. The tangential nucleus is a major avian vestibular nucleus whose principal cells are vestibular reflex projection neurons. From patch-clamp recordings on brain slices, the percentage of spontaneous spike firing principal cells, spike discharge rate, ionic conductances, and spontaneous excitatory postsynaptic currents (sEPSCs) were investigated one and three days after UVG. Already by one day after UVG, sEPSC frequency increased significantly on the lesion side, although no differences were detected in the percentage of spontaneous spike firing cells or discharge rate. In compensated chickens three days after UVG, the percentage of spontaneous spike firing cells increased on the lesion side and the discharge rate increased bilaterally. In uncompensated chickens three days after UVG, principal cells on the lesion side showed increased discharge rate and increased sEPSC frequency, whereas principal cells on the intact side were silent. Typically, silent principal cells exhibited smaller persistent sodium conductances and higher activation thresholds for the fast sodium channel than spiking cells. In addition, silent principal cells on the intact side of uncompensated chickens had larger dendrotoxin-sensitive potassium conductance, with a higher ratio of Kv1.1 surface/cytoplasmic expression. Increased sEPSC frequency in principal cells on the lesion side of uncompensated chickens was accompanied by decreased Kv1.2 immunolabeling of presynaptic terminals on principal cell bodies. Thus, both intrinsic ionic conductances and excitatory synaptic inputs play crucial roles at early stages after lesions. Unlike the principal cells in compensated chickens which showed similar percentages of spontaneous spike firing cells, discharge rates, and sEPSC frequencies bilaterally, principal cells in uncompensated chickens displayed gross asymmetry in these properties bilaterally.

Epidemiological studies show that increased plasma total homocysteine (tHcy) level was an independent risk factor of cardiovascular diseases. This study was aimed to investigate the relationship between tHcy level and prognosis of first-onset stroke in Chinese people. One hundred ninety six patients with first-onset ischemic stroke and ninety-five patients with first-onset hemorrhagic stroke were enrolled in this study. The patients were divided into two groups in terms of tHcy level (< 18 μmol/L and ⩾ 18 μmol/L). The plasma tHcy level was detected by a high performance liquid chromatography method with fluorescence detection. All the patients underwent a 5-year follow-up. Survival analysis shows that the probability of death or new vascular events in the ischemic stroke patients with high tHcy level (⩾ 18 μmol/L) was significantly higher than that in the counterparts with lower tHcy level (< 18 μmol/L) (50.9% and 28.7%, respectively, P = 0.004). The relative risk of death or new vascular events was 2.363 (95% CI, 1.209 to 4.617, P = 0.012) in ischemic stroke patients with high tHcy levels(⩾ 18 μmol/L) compared to those with a lower tHcy level (< 18 μmol/L). The increased tHcy level was significantly associated with the risk of death or new vascular events (OR, 2.492, 95% CI, 1.148 to 5.407, P = 0.021) in patients with ischemic stroke in the exclusion of the influence of other risk factors such as gender, age, body mass index, plasma cholesterol level, the history of hypertension, diabetes or smoking. However, in the patients with hemorrhagic stroke, there was no significant difference in the probability of death or new vascular events between patients with a high tHcy level and those with a lower tHcy level (33.3% and 28.2%, respectively, P = 0.546). Increased tHcy level was an independent risk factor for a worse outcome in patients with first-onset ischemic stroke, but not in hemorrhagic stroke patients.

The chick tangential nucleus is a major vestibular nucleus whose principal cells participate in the vestibular reflexes. During development, most mature vestibular nucleus neurons must acquire repetitive firing of action potentials on depolarization and spontaneous spike activity to process signals effectively. In the chicken, these properties emerge gradually in the embryo, starting the week before (E13, E16) and continuing through the first week after hatching (H7). Since gap junction-mediated cell coupling may influence the emergence of neuronal excitability, we investigated whether neuron-neuron and neuron-glia coupling are present in this morphologically distinctive vestibular nucleus during the period for establishing signal processing. In brain slices, principal cells were injected with biocytin in the whole-cell configuration and visualized via confocal imaging at E13, E16, and H7. The incidence of dye coupling between the injected principal cell and neurons was 42% at E13, 75% at E16, and 7% at H7, whereas the incidence of dye coupling with glia was 100% at both embryonic ages but decreased to 27% by H7. For each injected principal cell at E13, one coupled neuron and 35 coupled glia were detected, whereas three coupled neurons and 12 coupled glia were observed at E16, and few if any coupled neurons and glia were detected at H7. These results suggest that neuron-neuron and neuron-glia coupling are developmentally regulated and present before, but not after, the onset of mature signal processing by these neurons. Thus, transient neuron-neuron and neuron-glia coupling may both play roles in establishing excitability in vestibular nucleus neurons during development.

The modulation of GABA-gated ion channel responses to GABA, pentobarbital and diazepam by muscarine was studied in freshly isolated rat dorsal root ganglion neurons using a whole-cell patch-clamp technique. Muscarine enhanced current activated by 5 microM GABA dose-dependently with an EC50 of 40 +/- 2 microM. This potentiation was not blocked by pirenzepine, gallamine and atropine, the specific and non-specific muscarinic receptor antagonists. Muscarine shifted the GABA dose-response curve to the left, with the GABA EC50 decreased from 45 +/- 2 to 13 +/- 2 microM. The maximal response to GABA was suppressed to 89.3 +/- 4.6% as compared with the control (100%) by 80 microM muscarine. Muscarine potentiated GABA (1-100 microM)-activated current in a voltage-independent manner. Muscarine shifted the dose-response curve for pentobarbital enhancement of GABA-activated current to the left, and the enhancement of GABA-activated current by muscarine was additive to that of pentobarbital over all pentobarbital concentrations. Muscarine shifted the dose-response curve for diazepam (1-100 nM) enhancement of GABA-activated current to the left. However, muscarine attenuated the facilitatory effect of saturating concentrations of diazepam (100 nM). The potentiating effect of muscarine was blocked by 1 nM ethyl-beta-carboline-3-carboxylate, the inverse agonist of benzodiazepine receptors. These results suggest that GABA-gated ion channel responses to GABA and pentobarbital were potentiated by muscarine and the binding site(s) for muscarine might be related to those for diazepam.

Beta amyloid (Aβ)-induced oxidative stress and chronic inflammation in the brain are considered to be responsible for the pathogenesis of Alzheimer's disease (AD). Salidroside, the major active ingredient of Rhodiola crenulata, has been previously shown to have antioxidant and neuroprotective properties in vitro. The present study aimed to investigate the protective effects of salidroside on Aβ-induced cognitive impairment in vivo. Rats received intrahippocampal Aβ1-40 injection were treated with salidroside (25, 50 and 75 mg/kg p.o.) once daily for 21 days. Learning and memory performance were assessed in the Morris water maze (days 17-21). After behavioral testing, the rats were sacrificed and hippocampi were removed for biochemical assays (reactive oxygen species (ROS), superoxide dismutase (SOD), glutathione peroxidase (GPx), malondialdehyde (MDA), acetylcholinesterase (AChE), acetylcholine (ACh)) and molecular biological analysis (Cu/Zn-SOD, Mn-SOD, GPx, nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, nuclear factor κB (NF-κB), inhibitor of κB-alpha (IκBα), cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), receptor for advanced glycation end products (RAGE)). Our results confirmed that Aβ1-40 peptide caused learning and memory deficits in rats. Further analysis demonstrated that the NADPH oxidase-mediated oxidative stress was increased in Aβ1-40-injected rats. Furthermore, NF-κB was demonstrated to be activated in Aβ1-40-injected rats, and the COX-2, iNOS and RAGE expression were also induced by Aβ1-40. However, salidroside (50 and 75 mg/kg p.o.) reversed all the former alterations. Thus, the study indicates that salidroside may have a protective effect against AD via modulating oxidative stress and inflammatory mediators.

Principal cells of the chick tangential nucleus are vestibular nucleus neurons in the hindbrain. Although detailed information is available on the morphogenesis of principal cells and synaptogenesis of primary vestibular fibers, this is the first study of their early functional development, when vestibular terminals emerge at embryonic days 10 and 13 (E10 and E13). At E10, 60% of principal cells generated spikes on depolarization, whereas 50% exhibited excitatory postsynaptic currents (EPSCs) on vestibular-nerve stimulation. The frequency was 0.2 Hz for glutamatergic spontaneous EPSCs (sEPSCs) at −60 mV, and 0.6 Hz for spontaneous inhibitory postsynaptic current (sIPSC) at +10 mV and completely GABAergic. All of these synaptic events were TTX-insensitive, miniature events. At E13, 50% of principal cells generated spikes on depolarization and 82% exhibited EPSCs on vestibular-nerve stimulation. The frequency was 0.7 Hz for sEPSCs at −60 mV, and 0.8 Hz for sIPSCs at +10 mV. Most principal cells had sIPSCs composed of both GABAergic (75%) and glycinergic (25%) events, but a few cells had only GABAergic sIPSCs. TTX decreased the frequency of EPSCs by 12%, and the IPSCs by 17%. In summary, at E10, some principal cells generated immature spikes on depolarization and EPSCs on vestibular-nerve stimulation. At E10, GABAergic events predominated, AMPA events had low frequencies, and glycinergic activity was absent. By E13, glycinergic events first appeared. This data were compared systematically to that obtained from the late-term embryo and hatchling to reveal the long-term sequence of changes in synaptic events and excitability and offer a broader understanding of how the vestibular system is assembled during development.

Using water/AOT/n-octane reversed micelle as the medium, the optical signal of the reactive intermediate of laccase-catalyzed oxidation of o-phenylenediamine, which was indetectable in aqueous solutions, was successfully captured. Thus online kinetic studies of the intermediate were accomplished. Two-way kinetic spectral data were acquired with stopped-flow technique. By resolving the data with global analysis software, both the kinetic curves and the absorption spectra of the components involved in the reaction process were simultaneously obtained. The whole reaction in the reversed micelle was proved to be composed of two successive steps, an enzymatic generation of the intermediate and a following nonenzymatic decay of the intermediate. A consecutive first-order kinetic model of the whole reaction was confirmed. The influences of microenvironmental factors of the medium (such as the pH value of the water pool and the water/AOT ratio) on the detection of the intermediate were also investigated.

The principal cells of the chick tangential nucleus are second-order vestibular neurons involved in the vestibu- loocular and vestibulocollic reflexes. The spontaneous syn- aptic activity of morphologically identified principal cells was characterized in brain slices from 1-day-old hatchlings (H1) using whole-cell voltage-clamp recordings and Cs-gluconate pipet solution. The frequency was 1.45 Hz for spontaneous excitatory postsynaptic currents (sEPSCs) and 1.47 Hz for spontaneous inhibitory postsynaptic currents (sIPSCs). Us- ing specific neurotransmitter receptor antagonists, all of the sEPSCs were identified as AMPA receptor-mediated events, whereas 56% of the sIPSCs were glycine and 44% were GABAA receptor-mediated events. On exposure to TTX, the frequency of EPSCs decreased by 68%, while the frequency of IPSCs decreased by 33%, indicating greater EPSC depen- dency on presynaptic action potentials. These data on spon- taneous synaptic activity at H1 were compared with those obtained in previous studies of 16-day old embryos (E16). After birth, the spontaneous synaptic activity exhibited in- creased EPSC frequency, increased ratio for excitatory to inhibitory events, increased percentage of TTX-dependent EPSCs, and faster kinetics. In addition, the ratio for glycine/ GABA receptor-mediated events increased significantly. Al- together, these data indicate that at hatching spontaneous synaptic activity of vestibular nucleus neurons in brain slices of the chick tangential nucleus undergoes appreciable changes, with increased frequency of EPSCs and glycinergic activity playing more important roles compared with the late- term chick embryo when GABAergic activity prevailed. The definition of this developmental pattern of synaptic activity in vestibular nucleus neurons should contribute to understand- ing how vestibular reflex activity is established in the hatch- ling chick. © 2004 IBRO. Published by Elsevier Ltd. All rights reserved.

A sensitive, rapid and accurate analysis of the amino acid neurotransmitters in dorsal root ganglion (DRG) of rat was accomplished by capillary electrophoresis with a laser-induced fluorescence-charged coupled device (LIF-CCD) and fluorescein isothiocyanate (FITC) derivatization. Since the CCD is an image detector with a matrix array of photoelements, the photogenerated charge generated from single photoelements was summed up by on-chip charge binning, and used to improve the detection sensitivity. A self-compiled data processing program was used to calculate digitally the summation of the multiwavelength data and transform three-dimensional electropherograms into two-dimensional electropherograms. Use of this program improved the detection sensitivity, the extraction of useful statistical data and the performance of the quantitative analysis. Owing to its excellent detectability and the unique characteristic of its two-dimensional format, which allows to perform analog and digital summation, high detection sensitivity with detection limits ranging from 1.2 to 17.2 nM was obtained for the amino acid neurotransmitters by LIF-CCD. This method was demonstrated to be a powerful tool for analyzing the complex biological samples through quantitative determination of the six amino acid neurotransmitters in the DRG of rat.

A series of titanium complexes Ar[O, E]Cp*TiCl (Cp* = C5Me5, Ar = 1,2-phenylene, E = NH (1a); Ar = 1,2-phenylene, E = O (1b); Ar = 2,2′-diphenylene, E = O (1c); Ar = 2,2′-dinaphthalene, E = O (1d)) has been prepared by the reaction of corresponding phenol derivatives with Cp*TiCl3 in the presence of excessive triethylamine. Under the conditions of low Al/Ti molar ratio (e.g. 500) and high reaction temperatures (≥ 70°C), all the titanium complexes display higher catalytic activities towards the syndiospecific polymerization of styrene, in the presence of modified methylaluminoxane (MMAO) as a cocatalyst, than their mother complex Cp*TiCl3. The catalyst activities and polymer yields as well as polymer properties are considerably affected by the steric and electronical effects of the bidentate ligands.