Microsampling in Targeted Mass Spectrometry-Based Protein Analysis of Low-Abundance Proteins.

This paper presents a protocol with detailed descriptions for efficient sample cleanup of low-abundance proteins from dried samples. This is performed using bead-based proteolysis prior to proteotypic peptide affinity-capture and liquid chromatography tandem mass spectrometry (LC-MS/MS) determination. The procedure can be applied to both conventional dried samples using paper cards (e.g., dried blood spots [DBSs] and dried serum spots [DSSs]), as well as samples collected with newer sampling methods such as volumetric absorptive microsampling (VAMS). In addition to describing this procedure, the preparation of both trypsin beads and antibody-coated beads is presented in a step-by-step manner in this work. The advantages of the presented procedure are time-efficient proteolysis using beads and selective robust cleanup using peptide affinity-capture. The current procedure describes the determination of the low-abundance small-cell lung cancer (SCLC) biomarker, progastrin-releasing peptide (ProGRP), in dried serum (both DSSs and VAMS). Detailed procedures for bead preparation make it easier to implement the workflow in new applications or other laboratories. It is demonstrated that the results may be dependent on the sampling material; for the present project, higher signal intensities were seen for samples collected using VAMS compared to DSSs.

A multi-functional minimally-disruptive portable electrochemical system based on yeast/Co3O4/Au/SPEs for blood lead (II) measurement.

A minimally-disruptive portable electrochemical system is constructed by combining a hand-held syringe as reservoir with disposable screen-printed electrodes (SPEs) modified with a simple and efficient yeast/Co3O4/Au material for lead determination by a square-wave voltammetry (SWV) method. Not only can it preserve the operation and advantages of the conventional electrochemical procedure, but it also integrates sampling, filtering and analysis to make the determination of lead convenient and effective at higher and lower concentration levels. This is the first report of a microbial biosensor based on active yeast crosslinked to Co3O4/Au particles using glutaraldehyde as the crosslinking agent. The determination process is simplified by introducing a fiber filter and takes only 150 s with the developed system, which illustrates its simplicity, speed and detection accuracy. Also, the design shows a wide log-linear dynamic range (LDR) from 10-8 to 10-14 g·L-1, with a limit of detection (LOD) of 3.45 × 10-15 g·L-1 (S/N = 3). Additionally, the proposed system was used to determine lead in blood samples, which demonstrated the potential of this biosensor for use in practical applications. Furthermore, this study provides a basis for the development of microscale blood devices for lead measurement.

Capillarity-based preparation system for optical colorimetric sensor arrays.

In recent years, optical colorimetric sensor arrays have demonstrated beneficial features, including rapid response, high selectivity, and high specificity; as a result, it has been extensively applied in food inspection and chemical studies, among other fields. There are instruments in the current market available for the preparation of an optical colorimetric sensor array, but it lacks the corresponding research of the preparation mechanism. Therefore, in connection with the main features of this kind of sensor array such as consistency, based on the preparation method of contact spotting, combined with a capillary fluid model, Washburn equation, Laplace equation, etc., this paper develops a diffusion model of an optical colorimetric sensor array during its preparation and sets up an optical colorimetric sensor array preparation system based on this diffusion model. Finally, this paper compares and evaluates the sensor arrays prepared by the system and prepared manually in three aspects such as the quality of array point, response of array, and response result, and the results show that the performance index of the sensor array prepared by a system under this diffusion model is better than that of the sensor array of manual spotting, which meets the needs of the experiment.

Colorimetric detection of Cr (VI) based on the leaching of gold nanoparticles using a paper-based sensor.

Herein, we have developed a simple, sensitive and paper-based colorimetric sensor for the selective detection of Chromium (Ⅵ) ions (Cr (VI)). Silanization-titanium dioxide modified filter paper (STCP) was used to trap bovine serum albumin capped gold nanoparticles (BSA-Au NPs), leading to the fabrication of BSA-Au NPs decorated membrane (BSA-Au NPs/STCP). The BSA-Au NPs/STCP operated on the principle that BSA-Au NPs anchored on the STCP were gradually etched by Cr (VI) as the leaching process of gold in the presence of hydrobromic acid (HBr) and hence induced a visible color change. Under optimum conditions, the paper-based colorimetric sensor showed clear color change after reaction with Cr (VI) as well as with favorable selectivity to a variety of possible interfering counterparts. The amount-dependent colorimetric response was linearly correlated with the Cr (VI) concentrations ranging from 0.5µM to 50.0µM with a detection limit down to 280nM. Moreover, the developed cost-effective colorimetric sensor has been successfully applied to real environmental samples which demonstrated the potential for field applications.

Synthesis of maltodextrin-grafted-cinnamic acid and evaluation on its ability to stabilize anthocyanins via microencapsulation.

In this work, maltodextrin-grafted-cinnamic acid (MD-g-CA) was synthesised and used as wall material to improve the stability of purple sweet potato anthocyanins (PSPa) via microencapsualtion. MD-g-CA was prepared through esterification in a two-step convenient synthesis procedure and characterised using infra-red (IR) spectroscopy. The IR data indicated the typical ester carbonyl stretching at around 1721 cm-1. Moreover, MD-g-CA could give about 40% inhibition of DPPH radical and present excellent UV-absorption, which were notably better than that of native MD. Maltodextrin (MD) and MD-g-CA were used to prepare PSPa microcapsules by spray drying. The stability of PSPa was evaluated by UV-Vis analysis. The microcapsules produced by MD-g-CA showed a spheres-like appearance with some cracks. Storage tests revealed that the degradation rate of PSPa embedded by MD-g-CA was much lower than that of free PSPa under the same condition. Thus, MD-g-CA could be used as an effective wall material to improve stability of anthocyanins.

A novel device based on a fluorescent cross-responsive sensor array for detecting lung cancer related volatile organic compounds.

In this paper, a novel, simple, rapid, and low-cost detection device for lung cancer related Volatile Organic Compounds (VOCs) was constructed. For this task, a sensor array based on cross-responsive mechanism was designed. A special gas chamber was made to insure sensor array exposed to VOCs sufficiently and evenly, and FLUENT software was used to simulate the performance of the gas chamber. The data collection and processing system was used to detect fluorescent changes of the sensor arrays before and after reaction, and to extract unique patterns of the tested VOCs. Four selected VOCs, p-xylene, styrene, isoprene, and hexanal, were detected by the proposed device. Unsupervised pattern recognition methods, hierarchical cluster analysis and principal component analysis, were used to analyze data. The results showed that the methods could 100% discriminate the four VOCs. What is more, combined with artificial neural network, the correct rate of quantitative detection was up to 100%, and the device obtained responses at concentrations below 50 ppb. In conclusion, the proposed detection device showed excellent selectivity and discrimination ability for the VOCs related to lung cancer. Furthermore, our preliminary study demonstrated that the proposed detection device has brilliant potential application for early clinical diagnosis of lung cancer.

Development of a colorimetric sensor array for the discrimination of Chinese liquors based on selected volatile markers determined by GC-MS.

A new colorimetric sensor array was developed for the discrimination of 12 high-alcoholic Chinese base liquors from Luzhou Co., Ltd., and 15 commercial Chinese liquor of different brands as well as flavor types. Seventeen volatile compounds within four chemical groups were determined as markers in the base liquor by GC-MS analysis and factor analysis method (FAM). A specialized colorimetric sensor array composed of 20 sensitive dots was fabricated accordingly to obtain sensitive interaction with different types of volatile markers. Discrimination of the liquor samples was subsequently performed using chemometric and statistical methods, including principal component analysis (PCA) and hierarchical clustering analysis (HCA). The results suggested that facile identification of either base liquors with high-alcoholic volume or commercial liquors of the same flavor types could be achieved by analysis of the color change profiles. The response of the sensor improved significantly in comparison with those that rely on nonspecific interactions, and no misclassification was observed for both liquor samples using two chemometric methods. Besides, it was also found that the discrimination is closely related to the characteristic flavor compounds (esters, aldehydes, and acids) and alcoholic strength in liquors, and its performance was even comparable with that of GC-MS.

A novel chemical detector using colorimetric sensor array and pattern recognition methods for the concentration analysis of NH3.

With a colorimetric sensor array comprising chemoresponsive dyes, a simple, rapid, and cost-effective integrated system for differentiating low-concentration gases was described. The system could be used to identify gases by detecting the color change information of the chemoresponsive dyes based on porphyrins before and after reaction with the target gas; the colorimetric sensor array images were collected by a charge coupled device and processed with image analysis to get the color changes of the dyes in the array. Temperature, humidity, and flux of the chamber could be detected and displayed on the personal computer screen. A low-concentration [30-210 ppb (parts per 10(9))] NH(3) was detected by the system. This prototype successfully differentiated four concentration levels of NH(3) in less than 1 min. Pattern recognition methods, such as the backpropagation neural network and the radial basis function neural network, validated the effect of the developed sensor system both with 100% classification with feature vectors at single time point as inputs.

Effect of shuizhongcao granule on cellular immune function of experimental animal with recurrent aphthous stomatitis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of Shuizhongcao Granule (SZCG) on cellular immune function in animal model of recurrent aphthous stomatitis (RAS).</p><p><b>METHODS</b>Experimental animal model of RAS were established by immunological method. The modeled animals were randomized into 4 groups, Group A was the model control group, Group B was treated by Levamisole (2.5 mg/mL), Group C and D was treated with high-dose (0.5 g/mL) and low-dose (0.24 g/mL) SZCG respectively. The medication was administered via gastric perfusion, starting from the 7th week of the experiment, and continued for 28 days. Meanwhile, a normal control group was set up. Percentage of T-lymphocyte subsets, CD3, CD4 and CD8, and serum levels of interleukin 10 and 12 (IL-10 and IL-12) in peripheral blood were detected before and after treatment.</p><p><b>RESULTS</b>Percentages of CD3, CD4 and CD4/CD8 ratio in the model control group were 40.50 +/- 7.46%, 30.80 +/- 5.33% and 79.56 +/- 8.32 respectively, all lower than those in the normal control group (P<0.05); level of serum IL-10 was higher (8.02 +/- 0.53 ng/L) and that of IL-12 was lower (15.51 +/- 1.35 ng/L) in the model animal than those in normal control (P<0.05). After treatment, levels of CD3, CD4, CD4/CD8 and IL-12 in Group B were 55.30 +/- 6.33%, 43.50 +/- 5.33%, 99.29 +/- 13.84 and 17.72 +/- 1.70 ng/L, those in Group C were 61.50 +/- 4.32%, 47.20 +/- 4.37%, 103.30 +/- 7.42 and 18.18 +/- 1.54 ng/L, and in Group D were 58.50 +/- 6.03%, 42.80 +/- 3.17%, 110.27 +/- 12.85 and 17.74 +/- 1.96 ng/L, respectively, all were higher than those in Group A, but levels of IL-10 in the three treatment groups were lower than that in Goup A (P<0.05 or P<0.01).</p><p><b>CONCLUSION</b>The immune function of RAS model animals is in the suppressed condition; SZCG can improve the immune suppression to achieve its therapeutic effect on the disease.</p>

Evaluation for disease-specific quality of life in children with sleep-disordered breathing / 中华全科医师杂志

Objective To evaluate the impact of sleep-disordered breathing(SDB)on the quality of life(QOL)in children with SDB and investigate the value of tonsillectomy and adenoidectomy in improving their QOL.Methods Questionnaire of disease-specific quality of life for children with obstructive sleep apnea with 18 items(OSA-18)was used to evaluate QOL in 168 children with SDB within four weeks before and six to 12 months after surgical operation,and correlation between OSA-18 scores to apnea hypopnea index(AHI)and minimum oxygen saturation(MiO2)in 62 of them was evaluated with polysomnography(PSG),with 50 healthy children without snoring as controls.Results Overall scores of OSA-18 in children of SDB group before surgical operation were significantly higher than those in children of control group(t=15.12,P＜0.01).and QOL was seriously affected in 30.4 percent of the children with SDB.Scores of OSA-18 correlated to AHI and MiO2 before surgical operation.QOL was improved significantly in 73.8 percent of children with SDB after tonsillectomy and adenoidectomy.Conclusions SDB has obvious impact on pediatric QOL,which can be improved by tonsillectomy and adenoidectomy.There exists significant correlation between OSA-18 scores and objective indicators of PSG,and the former can be used as clinical diagnosis for SDB in children and as quantitative evaluation for the effectiveness of intervention.

Supramolecular Polypseudorotaxanes composed of star-shaped porphyrin-cored poly(epsilon-caprolactone) and alpha-cyclodextrin.

Star-shaped porphyrin-cored poly(epsilon-caprolactone) (SPPCL) was synthesized using a tetrahydroxyethyl-terminated porphyrin as a core initiator and stannous octoate as a catalyst in bulk at 120 degrees C. The molecular weight of as-synthesized polymer could be adjusted linearly by controlling the molar ratio of epsilon-caprolactone to porphyrin core initiator, and the molecular weight distribution was reasonably narrow. Supramolecular polypseudorotaxanes were prepared by inclusion complexation of SPPCL with alpha-cyclodextrin (alpha-CD) and thoroughly characterized by means of FT-IR, 1H NMR, 13C CP/MAS NMR, DSC, TGA, and WAXD. The results demonstrated that the porphyrin-cored polypseudorotaxanes formed through alpha-CD molecules threading onto the branch chains of star-shaped SPPCL polymers, and they had a channel-type crystalline structure. Meanwhile, the original crystallization of SPPCL polymers within the polypseudorotaxanes was completely suppressed in the alpha-CD cavities. Moreover, inclusion complexation between SPPCL and alpha-CD enhanced the thermal stability of both the guest SPPCL polymers and the host alpha-CD. Furthermore, both the SPPCL polymers and the polypseudorotaxanes showed similar fluorescent and UV-vis spectra compared with porphyrin core initiator. Consequently, this will not only provide potentially porphyrin-cored poly(epsilon-caprolactone) and its polypseudorotaxanes for photodynamic therapy but also improve the compatibility between poly(epsilon-caprolactone) and peptide drugs for drug delivery.

mRNA expression change of myelin gene in hippocampus following cerebral ischemia / 中华急诊医学杂志

Objective To study the mRNA expression of myelin basic protein (MBP) and myelin oligodendregha glyeoprotein (MOG) in hippocampus of rats following global brain ischemia.Method The four- vessel occlusion animal model in the Sprague-Dawley rats was used in this study.The mRNA expression levels of MBP and MOG in the hippocampus of rats were analyzed by reverse transcription polymerase chain reaction (RT- PCR) at day 2,4,7,14 and 28 days after global brain ischemia.There were eight rats at each time-point and sham operated group.Results The mRNA expression of both MBP and MOG in hippocampus of rats decreased at 2 days after global brain ischemia.The gene expression of myelin gene decreased significantly at 7 days and it reached to the lowest level at 28 days.Compared with sham operated group,the gene expression of MBP and MOG in hippocampus of rats decreased significantly at 7,14 and 28 days after global brain ischemia (P

Prediction meaning of daytime sleepiness evaluation for patiens with sleep apnea sydrome combined with cerebral vascular disease occurrence / 临床神经病学杂志

Objective To investigate whether scores of epworth sleepiness scale (ESS) assessing daytime sleepiness were associated with sleep apnea syndrome(SAS)combined with cerebral vascular disease and ESS could be used to predict cerebral vascular disease occurrence.Methods A group of 62 SAS combined with transient ischemic attack(TIA), a group of 53 SAS with combined hypertension and the other group of 61 SAS combined without other diseases were monitored by polysomnography(PSG), daytime sleepiness of these SAS patients were assessed by ESS at the same time. Results Compared to SAS combined without other diseases, those SAS combined with TIA and hypertension patients had significant higher scores of ESS,apnea/hypopnea index (AHI) and micro-arousal index(MAI)(all P