Improving the Signal-to-Noise Ratio of Axial Displacement Measurements of Microspheres Based on Compound Digital Holography Microscopy Combined with the Reconstruction Centering Method.

In 3D microsphere tracking, unlike in-plane motion that can be measured directly by a microscope, axial displacements are resolved by optical interference or a diffraction model. As a result, the axial results are affected by the environmental noise. The immunity to environmental noise increases with measurement accuracy and the signal-to-noise ratio (SNR). In compound digital holography microscopy (CDHM)-based measurements, precise identification of the tracking marker is critical to ensuring measurement precision. The reconstruction centering method (RCM) was proposed to suppress the drawbacks caused by installation errors and, at the same time, improve the correct identification of the tracking marker. The reconstructed center is considered to be the center of the microsphere, rather than the center of imaging in conventional digital holographic microscopy. This method was verified by simulation of rays tracing through microspheres and axial moving experiments. The axial displacements of silica microspheres with diameters of 5 µm and 10 µm were tested by CDHM in combination with the RCM. As a result, the SNR of the proposed method was improved by around 30%. In addition, the method was successfully applied to axial displacement measurements of overlapped microspheres with a resolution of 2 nm.

Intermolecular Triazene Alkene Cycloaddition via Lewis Base Catalysis: Access to Diverse Trifluoromethylated Pyrazolines.

A novel approach to chemoselective synthesis of biologically important CF3 -subsituted pyrazolines was developed via a Lewis base catalyzed intermolecular triazene cycloaddition reaction of an array of terminal/internal alkenes with CF3 CHN2 . This strategy features a catalytic amount of 1,8-diazabicyclo[5.4.0]undec-7-ene, high yields (up to 95 %), wide substrate scope and excellent functional group tolerance (54 examples). Importantly, we preformed scaffold diversification of a panel of known pharmaceuticals, natural products, and bioactive heterocycles to generate the corresponding pyrazoline derivatives with potential broad bioactivities for further development.

Effect of water change on quality deterioration of Pacific white shrimp (Litopenaeus vannamei) during partial freezing storage.

The correlation between water changes and quality deterioration of Litopenaeus vannamei during partial freezing storage was evaluated in this study. Significant increases in cross-sectional area and equivalent diameter are detected, but the roundness and longiness of the ice crystals show irregular growth. Within the extension of storage, the bound water (T2b) and immobilized water (T21) decreased significantly. However, the free water (T22) increased significantly. Quality determination showed significant decrease in total sulfhydryl and Ca2+-ATPase, but significant increase in disulfide bonds during storage. Correlation analysis revealed that cross-sectional area showed significant negative correlation with total sulfhydryl and Ca2+-ATPase, while significant positive correlation with disulfide bonds, respectively. The correlation between water distribution index and Ca2+-ATPase, disulfide bonds was significant, respectively. Predicted models for the growth of ice crystals with respect to cross-sectional area and equivalent diameter size have been developed with the help of the Arrhenius model.

Uric acid promotes myocardial infarction injury via activating pyrin domain-containing 3 inflammasome and reactive oxygen species/transient receptor potential melastatin 2/Ca2+pathway.

Cardiomyocytes injury has been considered as a key contributor for myocardial infarction (MI). Uric acid (UA) can induce cardiomyocytes injury, which is closely related to NLRP3 activation and inflammatory factor generation. However, the mechanism how UA modulates cardiomyocytes remains elusive. Western blotting and qRT-PCR were applied for measuring protein and mRNA expression, respectively. ROS production and Ca2+ influx were measured by flow cytometry. Patch clamp technique was used for measuring transient receptor potential melastatin 2 (TRPM2) channel. Ligation of left anterior descending for 2 h was performed to induce MI animal model. The rats were treated by different concentration of uric acid. The artery tissues were stained by HE and collected for measurement of NLRP3 and inflammatory factors. Supplementation of UA significantly promoted apoptosis, and augmented the expression of intercellular adhesion molecule-1, chemoattractant protein-1, vascular cell adhesion molecule-1, and NLRP3 inflammasome. Knockdown of NLRP3 reversed the influence of UA on MI by decreasing collagen deposition, fibrotic area, apoptosis. The expression of NLRP3 inflammasome increased markedly after treatment of UA. UA activated ROS/TRPM2/Ca2+ pathway through targeting NLRP3. UA activated NLRP3 inflammasome and augments inflammatory factor production, which in turn exacerbates cardiomyocytes injury. Knockdown of NLRP3 reversed the influence of UA on apoptosis and cell cycle. UA may promote cardiomyocytes injury through activating NLRP3 inflammasome and ROS/TRPM2 channel/Ca2+ pathway.

Identification of adulterated milk based on auto-correlation spectra.

A qualitative analysis of melamine-adulterated milk was proposed based on two-trace two-dimensional (2T2D) auto-correlation spectra. The concentration of melamine was used as external perturbation, and 40 adulterated samples of each brand with different concentrations of melamine (0.01 g/L to 1 g/L) were configured. Four brands of milk were used to configure experimental samples, including Guangming brand, Mengniu brand, Sanyuan brand and Wandashan brand. Spectroscopic data of pure milk and melamine-adulterated milk were measured by infrared (IR) (80-4000 cm-1) spectrophotometer. 2T2D auto-correlation spectral technology combined with least squares support vector machine (LS-SVM) method was used for qualitative analysis. The two strongest auto-correlation peaks in the auto-correlation spectra were selected for modeling. For Guangming brand, the intensities of auto-correlation at two wave numbers 2898 cm-1 and 2972 cm-1 were selected as independent variables. For Mengniu brand, the intensities of auto-correlation at two wave numbers 2852 cm-1 and 2920 cm-1 were selected. For Sanyuan brand, the intensities of auto-correlation at two wave numbers 2900 cm-1 and 2974 cm-1 were selected. For Wandashan brand, the intensities of auto-correlation at two wave numbers 2900 cm-1 and 2974 cm-1 were selected. For four brands fused together, the intensities of auto-correlation at two wave numbers 2900 cm-1 and 2974 cm-1 were selected. For each brand, the accuracy of qualitative analysis was 100 %. For four brands fused together, the accuracy of qualitative analysis was 99.05 %. In this way, it greatly reduced the amount of data to be processed. This study showed that 2T2D auto-correlation spectral technology combined with LS-SVM method was perfect for the discrimination of melamine-adulterated milk.

Temperature-perturbed two-dimensional generalized correlation characteristic slice spectra combined with multivariate method to identify adulterated milk.

In order to improve the discrimination accuracy of adulterated milk, a detection method was proposed based on temperature-perturbed generalized two-dimensional (2D) correlation characteristic slice spectra. A total of 240 samples were prepared including three brands of 40 pure milk and 40 urea-tainted milk, respectively. The infrared attenuated total reflection spectra of each sample were collected at different temperatures. Synchronous 2D infrared correlation spectrum of each sample was calculated under the external perturbation of temperature. The characteristic slice spectra of each sample were extracted from synchronous 2D correlation spectrum at characteristic peaks of milk and adulterants. N-way partial least squares discriminant analysis (NPLS-DA) models of single brand and the fusion of three brands of adulterated milk were established based on 2D correlation characteristics slice spectra. For comparison, the discrimination models were established using synchronous 2D correlation spectra and one-dimensional (1D) infrared spectra at room temperature, respectively. For the three brand fusion models, the discrimination accuracies of unknown samples were 100%, 98.8% and 82.7% using 2D correlation characteristic slice spectra, 2D correlation spectra, and 1D spectra, respectively. The results showed that the proposed method not only compressed the data, but also effectively extracted the characteristic information, and improved the accuracy of discrimination.

Slice spectra approach to synchronous Two-dimensional correlation spectroscopy analysis for milk adulteration discriminate.

The discrimination approach of adulterated milk was proposed combined synchronous two-trace two-dimensional (2T2D) correlation slice spectra at the characteristic wavebands of adulterant in milk with multivariate method. Two common adulterants, melamine and urea, were analyzed to demonstrate useful by the method. 2T2D (near infrared) NIR slice spectra at characteristic wavebands of adulterant were extracted from the synchronous 2T2D correlation spectra, and were input to construct the N-way partial least squares discriminant analysis (NPLS-DA) models. One-dimensional (1D) spectroscopy featuring all the present components in the samples combined with partial least squares discriminant analysis (PLS-DA) was also evaluated for comparison. The results indicated that for one kind of adulterant in model, prediction accuracies of slice spectral models were both 100% for melamine-adulterated and urea-adulterated samples discrimination. Moreover, for two kinds of adulterants in model, prediction accuracies of slice spectral models were 90.57% and 100% for melamine-adulterated and urea-adulterated discrimination, respectively, which was better than those of 1D whole models based on PLS-DA (only 81.13% and 98.15%, respectively). The comparison informs that the 2T2D slice spectra extracted at the characteristic wavebands of adulterant highlighted the adulterant spectral features and was obviously advantage to improve the discrimination accuracy. Meanwhile, the complexity of slice spectra is significantly reduced compared with the whole matrix of synchronous 2T2D correlation spectra.

Discrimination of adulterated milk using temperature-perturbed two-dimensional infrared correlation spectroscopy and multivariate analysis.

The discrimination method for adulterated milk is proposed based on temperature-perturbed two-dimensional (2D) infrared correlation spectroscopy and N-way partial least squares discriminant analysis (NPLS-DA). Two brands of pure and adulterated milk samples were prepared. The mid-infrared spectra of all samples were obtained from 30 â„ƒ to 55 â„ƒ with an interval of 5 â„ƒ. Under the perturbation of temperature, synchronous 2D correlation spectra were calculated to build discrimination models of pure milk and adulterated milk. In comparison, the NPLS-DA models were built based on three-dimensional (3D) stacked map (sample × temperature × wavenumber variable). For the NPLS-DA models of two brands of milk, the discrimination accuracy of unknown samples in the prediction set is 100% using temperature-perturbed 2D infrared correlation spectra, versus 77.8% using conventional 3D stacked map. The proposed method can be used as an alternative way for classifying pure and adulterated milk.

Comparative study of pyrochar and hydrochar on peanut seedling growth in a coastal salt-affected soil of Yellow River Delta, China.

Biochar (i.e., pyrochar and hydrochar) application is a promising strategy to improve soil quality and productivity. However, the comparison of biochars with different carbonization methods and feedstocks for the plant growth in the coastal salt-affected soil remains limited. In this study, a 30-day microcosmic experiment was conducted to compare the effects of pyrochars and hydrochars derived from reed straw (RPC and RHC) and cow manure (CPC and CHC) on the peanut (Arachis hypogaea L.) seedling growth in a coastal salt-affected soil of Yellow River Delta, China. The results showed that RPC, CHC and CPC significantly elevated fresh shoot weight by 67.77%-89.37%, whereas the RHC amendment showed little effect. The malondialdehyde contents in peanut seedling leaves were significantly declined by 25.28%-35.51% with pyrochar and hydrochar amendments, which might be associated with the enhanced proline contents and K/Na ratios. The stimulation of certain phytohormones (i.e., indole-3-acetic acid, zeatin riboside, gibberellic acid 3) in peanut seedlings with pyrochar and hydrochar amendments might be attributed to the growth enhancement. RPC, CPC and CHC improved the soil properties and fertility such as cation-exchange capacity (CEC), total nitrogen, and available potassium and water holding capacity (WHC) of the coastal salt-affected soil. However, RHC not only significantly decreased soil CEC and WHC, but also increased soil exchangeable sodium percentage. The abundances of soil beneficial bacteria, such as f_Gemmatimonadacea, Sphingomonas, Blastococcus and Lysobacter were enhanced by RPC, CHC and CPC amendments, which were mainly associated with the increased WHC and CEC. Fungal community was less sensitive to pyrochar and hydrochar amendments than bacterial community according to the relative abundance and diversity, and beneficial fungi, such as Oidiodendron and Sarocladium were enriched in the CHC soil. Overall, the application of RPC, CHC and CPC showed greater potentials for the enhancement of peanut growth in a coastal salt-affected soil.

Toward a Practical Impedimetric Biosensor: A Micro-Gap Parallel Plate Electrode Structure That Suppresses Unexpected Device-to-Device Variations.

We propose a rational electrode design concept for affinity biosensors based on electrochemical impedance spectroscopy to substantially suppress unexpected device-to-device variations. On the basis that the uniformity of the current distribution affects the variation, a novel micro-gap parallel plate electrode (PPE) was developed, where two planar electrodes with edges covered with a SiO2 layer were placed face to face. The structure provides a uniform current distribution over the planar electrode surface and maximizes the contribution of the planar electrode surface to sensing. For a comparative study, we also fabricated a micro-structured interdigitated electrode (IDE) that has been widely adopted for high-sensitivity measurement, although its current is highly concentrated on the electrode edge corner. Protein G (PrG) molecules were immobilized on both electrodes to prepare an immunoglobulin G (IgG) biosensor on which the specific binding of PrG-IgG can occur. We demonstrated that the IgG sensor with the PPE has small device-to-device variations, in strong contrast to the sensor with the IDE having large device-to-device variations. The results indicate that the current distribution on the electrode surface is important to fabricating electrochemical impedance spectroscopy biosensors with small device-to-device variations. Furthermore, it was found that the PPE allows ultrasensitive detection, that is, the sensor exhibited a linear range from 1 × 10-13 to 1 × 10-7 mol/L with a detection limit of 1 × 10-14 mol/L, which is a record sensitivity at low concentrations for EIS-based IgG sensors.

Influence of brands on a discrimination model for adulterated milk based on asynchronous two-dimensional correlation spectroscopy slice spectra.

To improve the robustness of near infrared (NIR) identification models for the milk adulteration, a novel approach was explored based on asynchronous two-dimensional correlation spectroscopy (2D-COS) slice spectra obtained at characteristic wavebands for pure milk and adulterant combined with an N-way partial least squares discriminant analysis (NPLS-DA). NIR diffuse reflectance spectra from four different brands, Guangming (GM), Mengniu (MN), Sanyuan (SY), and Wandashan (WDS), were collected in range of 11,000 to 4000 cm-1. Influence of brands on discrimination models for adulterated milk was analyzed. The asynchronous 2D-COS slice spectra at 10 characteristics wavebands, including 4 wavebands for pure milk and 6 wavebands for urea, were input into NPLS-DA to construct discriminant models. External validations using five independent calibration sets from intrabrand or interbrand were established. The same prediction set of 26 SY samples was used to assess the prediction ability of different calibration sets and compared with traditional one-dimensional (1D) NIR spectra based on a partial least squares discriminant analysis (PLS-DA). It showed that for intrabrand model, the correct rates for the calibration and predication sets were 100% and 96.15%, respectively. For the interbrand model, the correct rates by the NPLS-DA for the calibration set of GM, MN, and WDS milk were both 100%. The corresponding rates for the prediction set were 73%, 88.46% and 69.23%, respectively, which were much higher than those of PLS-DA (only 50%, 53.83% and 50%, respectively). It was proven that model robustness was sensitive to changes in the milk brands. The proposed method can effectively reduce the influence of brands on the discrimination models.

Intermittent pneumatic compression (IPC) combined with an electric stimulator in the prevention of venous thromboembolism in stroke patients.

OBJECTIVE: To explore the clinical value of intermittent pneumatic compression (IPC) combined with an electric stimulator in the prevention of venous thromboembolism in stroke patients. METHODS: 58 stroke patients with hemiplegia admitted to the Department of Neurology in our hospital were recruited as the study cohort and randomly placed into a control group or an observation group, with 29 patients in each group. The control group was administered routine care and IPC, and the observation group was administered electric stimulation in addition to the treatment administered to the control group. We conducted a comparison and an analysis of the occurrences of thrombosis, the blood rheology indexes, the femoral vein flow rates, and the nursing satisfaction levels in the two groups. The circumferences of the hemiplegia patients' lower extremities were measured and recorded, and the circumferences of the healthy sides and the affected limbs were compared. RESULTS: On the 7th day after the intervention, the observation group had a higher incidence of deep vein thrombosis (DVT) than the control group (6.90% vs. 31.03%, P<0.05). The hemorheology indexes were lower after the treatment, and the hemorheology indexes in the observation group were higher compared with the control group (P<0.05). The observation group had a higher femoral vein flow velocity than the control group (P<0.05). On the 7th and 14th days after the intervention, the peak flow and average flow velocities in the observation group exceeded those of the control group (P<0.05). The nursing satisfaction rate in the observation group was higher than it was in the control group (96.55% vs. 82.76%, P<0.05). After 7 and 14 days of treatment, smaller changes in the hemiplegic limbs of the observation group were observed, compared to the control group (P<0.05). CONCLUSION: IPC combined with an electrical stimulator can enhance the patients' blood hypercoagulability, effectively prevent the occurrence of DVT, and improve the nursing satisfaction levels.

A Poly-Chitosan and Cis-Platinum Conjugated Composite Nanoparticle System for Liver Cancer Therapy.

The aim of this study was to test an effective nano-pole capsule loaded cis-platinum (CP) transplantation device for liver cancer (LC) therapy. A novel nano-pole capsule was designed as a new vector for storing CP. HepG2 cells and a B6/J mouse model were used to test the efficiency of polyethyleneimine-cis-platinum (PEI-CP) and poly-chitosan-cis-platinum (PC-CP). Infiltration efficiency and transplantation efficiency tests were performed to study the performance of the delivery system, and fibroblast reactions and macrophage numbers were observed, to test for immune rejection and foreign body reactions. The apoptosis rate and tumor diameter of hepatocellular carcinoma cells were used to evaluate the effect of the tumor therapy. We also studied the functional mechanism of different CP delivery systems. The infiltration and transplantation efficiencies of PC-CP were higher than that of PEI-CP; Less foreign body reaction appeared in PC system, with less fibroblast reaction and lower macrophage reaction. The clinical efficacy of PC-CP in terms of tumor apoptosis and diameter reduction was superior to that of PEI-CP. We demonstrated that PC-CP had a more significant alteration effect on mTOR, P-Ak, LC3 and P53. The PC system can better deliver and release drugs than PEI-CP, and may be a better choice for LC therapy in the future.

Preparation and characterization of biochar derived from co-pyrolysis of Enteromorpha prolifera and corn straw and its potential as a soil amendment.

Single biomass feedstock approach may not meet the requirements for developing biochar with desired characteristics for use as soil amendment. In this study, biochars were prepared by co-pyrolysis of nutrients-rich Enteromorpha prolifera and lignocellulose-rich corn straw (CPECs) at different mass ratios (3:7, 1:1, and 7:3). CPECs presented higher water-soluble N/P contents than corn straw biochar, and exhibited larger surface area, low Na content, and slower nutrient release rate than Enteromorpha prolifera biochar. The modification in physicochemical and properties of CPECs enhanced its potential application as a soil amendment. A pot experiment showed that CPECs derived from co-pyrolysis of appropriate ratios of Enteromorpha prolifera and corn straw (1:1, 7:3) significantly increased the biomass of cherry tomato plant by 64.05%, 40.03% and 81.88%, 55.25%, when compared with corn straw biochar and Enteromorpha prolifera biochar, respectively. The positive effects of CPECs were primarily attributed to improved soil properties (e.g., water holding capacity, soil organic matter, pH, soil nutrients content) and increased total N/P uptake by plants. The results of this work provided potentials of developing "designer" biochars to meet the multiple soil requirements by co-pyrolysis.

Protective effects of quercetin on traumatic brain injury induced inflammation and oxidative stress in cortex through activating Nrf2/HO-1 pathway.

BACKGROUND: Traumatic brain injury (TBI) has been a serious public health issue. Clinically, there is an urgent need for agents to ameliorate the neuroinflammation and oxidative stress induced by TBI. Our previous research has demonstrated that quercetin could protect the neurological function. However, the detailed mechanism underlying this process remains poorly understood. OBJECTIVE: This research was designed to investigate the mechanisms of quercetin to protect the cortical neurons. METHODS: A modified weight-drop device was used for the TBI model. 5, 20 or 50 mg/kg quercetin was injected intraperitoneally to rats at 0.5, 12 and 24 h post TBI. Rats were sacrificed three days post injury and their cerebral cortex was obtained from the injured side. The rats were randomly assigned into three groups of equal number: TBI and quercetin group, TBI group, and Sham group. The brain water content was calculated to estimate the brain damage induced by TBI. Immunohistochemical and Western blot assays were utilized to investigate the neurobehavioral status. Enzyme-linked immunosorbent assay and reverse transcription polymerase chain reaction were performed to evaluate the inflammatory responses. The cortical oxidative stress was measured by estimating the activities of malondialdehyde, superoxide dismutase, catalase and glutathione-Px. Western blot was utilized to evaluate the expression of nuclear factor erythroid 2-related factor 2 (Nrf-2) and heme oxygenase 1 (HO-1). RESULTS: Quercetin attenuated the brain edema and microgliosis in TBI rats. Quercetin treatment attenuated cortical inflammatory responses and oxidative stress induced by TBI insults. Quercetin treatment activated the cortical Nrf2/HO-1 pathway in TBI rats. CONCLUSIONS: Quercetin ameliorated the TBI-induced neuroinflammation and oxidative stress in the cortex through activating the Nrf2/HO-1 pathway.

The GLP-1 receptor herbal agonist morroniside attenuates neuropathic pain via spinal microglial expression of IL-10 and ß-endorphin.

OBJECTIVES: To assess the protective effect of the glucagon-like peptide-1 receptor (GLP-1R) agonist morroniside against neuropathic pain and its downstream mechanisms of activating microglial GLP-1R/interleukin-10 (IL-10)/ß-endorphin antinociceptive pathway. METHODS: Spinal nerve ligation-induced neuropathic pain rats were intrathecally injected with morroniside, with mechanical paw withdrawal threshold being assessed. The expression of spinal and cultured microglia IL-10 and ß-endorphin were detected with qRT-PCR. KEY FINDINGS: Morroniside alleviated mechanical allodynia in neuropathic rats, which was blocked by inhibiting or depleting microglia. In addition, neutralizing spinal IL-10 or ß-endorphin with specialized antibodies or blocking the µ-opioid receptor was able to fully reverse the morroniside-induced mechanical antiallodynia. Morroniside treatment stimulated the gene expression of IL-10 and ß-endorphin in the spinal lumbar enlargements of neuropathic rats as well as in primary cultured microglia. Furthermore, pretreatment with the IL-10 antibody blocked morroniside-stimulated ß-endorphin expression in the spinal cords of neuropathic rats and cultured primary microglia, whereas the ß-endorphin antibody failed to affect morroniside-stimulated gene expression of IL-10. CONCLUSIONS: These results reveal that morroniside produces therapeutic effects in neuropathy through spinal microglial expression of IL-10 and subsequent ß-endorphin after activation of GLP-1R.

The spinal microglial IL-10/ß-endorphin pathway accounts for cinobufagin-induced mechanical antiallodynia in bone cancer pain following activation of α7-nicotinic acetylcholine receptors.

BACKGROUND: Cinobufagin is the major bufadienolide of Bufonis venenum (Chansu), which has been traditionally used for the treatment of chronic pain especially cancer pain. The current study aimed to evaluate its antinociceptive effects in bone cancer pain and explore the underlying mechanisms. METHODS: Rat bone cancer model was used in this study. The withdrawal threshold evoked by stimulation of the hindpaw was determined using a 2290 CE electrical von Frey hair. The ß-endorphin and IL-10 levels were measured in the spinal cord and cultured primary microglia, astrocytes, and neurons. RESULTS: Cinobufagin, given intrathecally, dose-dependently attenuated mechanical allodynia in bone cancer pain rats, with the projected Emax of 90% MPE and ED50 of 6.4 µg. Intrathecal cinobufagin also stimulated the gene and protein expression of IL-10 and ß-endorphin (but not dynorphin A) in the spinal cords of bone cancer pain rats. In addition, treatment with cinobufagin in cultured primary spinal microglia but not astrocytes or neurons stimulated the mRNA and protein expression of IL-10 and ß-endorphin, which was prevented by the pretreatment with the IL-10 antibody but not ß-endorphin antiserum. Furthermore, spinal cinobufagin-induced mechanical antiallodynia was inhibited by the pretreatment with intrathecal injection of the microglial inhibitor minocycline, IL-10 antibody, ß-endorphin antiserum and specific µ-opioid receptor antagonist CTAP. Lastly, cinobufagin- and the specific α-7 nicotinic acetylcholine receptor (α7-nAChR) agonist PHA-543613-induced microglial gene expression of IL-10/ß-endorphin and mechanical antiallodynia in bone cancer pain were blocked by the pretreatment with the specific α7-nAChR antagonist methyllycaconitine. CONCLUSIONS: Our results illustrate that cinobufagin produces mechanical antiallodynia in bone cancer pain through spinal microglial expression of IL-10 and subsequent ß-endorphin following activation of α7-nAChRs. Our results also highlight the broad significance of the recently uncovered spinal microglial IL-10/ß-endorphin pathway in antinociception.

Low frequency electroacupuncture alleviates neuropathic pain by activation of spinal microglial IL-10/ß-endorphin pathway.

Electroacupuncture produces analgesia in chronic pain patients and animal models of pain hypersensitivity. The current study aims to illustrate the mechanisms underlying electroacupuncture-attenuated neuropathic pain. Neuropathic rats, induced by tight ligation of L5/L6 spinal nerves, markedly reduced mechanical thresholds in the ipsilateral hindpaws relative to the contralateral hindpaws. Low frequency (2 Hz) electroacupuncture stimulation for a period of 20 min alleviated neuropathic pain in the ipsilateral hindpaws of neuropathic rats in a time-dependent manner. The same electroacupuncture treatment also stimulated spinal gene and protein expression of IL-10 and ß-endorphin but not dynorphin A, measured by real-time quantitative PCR and ELISA kits. Intrathecal injection of the specific IL-10 antibody in neuropathic rats completely blocked electroacupuncture-increased spinal expression of ß-endorphin, but the ß-endorphin antibody failed to alter electroacupuncture-stimulated spinal IL-10 expression. Using a double fluorescence immunostaining technique, we observed that electroacupuncture stimulated spinal IL-10 and ß-endorphin expression in microglia but not in neurons or astrocytes in the spinal dorsal horn of neuropathic rats. Pretreatment with intrathecal injection of the microglial inhibitor minocycline, specific IL-10 antibody and ß-endorphin antiserum (but not the dynorphin A antibody), or selective µ-opioid receptor antagonist CTAP (but not κ- or δ-opioid receptor antagonist) completely blocked electroacupuncture-induced attenuation of neuropathic pain. These results suggest that low frequency electroacupuncture alleviates neuropathic pain through stimulation of the spinal microglial expression of IL-10 and subsequent expression of ß-endorphin.

Engineered bioactive nanoparticles incorporated biofunctionalized ECM/silk proteins based cardiac patches combined with MSCs for the repair of myocardial infarction: In vitro and in vivo evaluations.

The development of cardiac patches by the combination of bioactive nano- and bio-materials with mesenchymal stem cells signifies an auspicious approach for the treatment of cardiac repair in myocardial infarction. In the present investigation, we study about the cardiac function of morphology improved gold nanoparticles combined with extracellular matrix/silk proteins for the cell proliferation and expansion of cardiomyocytes. The physico-chemical and morphological characteristics demonstrated that spherical and homogeneous Au particles are distributed on the matrix porous surface for providing favorable conductivity and biological influences in cardiac repair. The in vitro cell studies of prepared patches have established enhanced cell compatibility and retention of cardiomyocytes survival. The in vivo determinations imply that Au-ESF group decreases infarct size to 65% from 89% in control group. These developed cardiac patches can be highly suitable in the cardiac regeneration and offer new platform in cardiac tissue engineering.

Involvement of d-amino acid oxidase in cerebral ischaemia induced by transient occlusion of the middle cerebral artery in mice.

BACKGROUND AND PURPOSE: d-Amino acid oxidase (DAAO) is a flavine adenine dinucleotide-containing flavoenzyme and specifically catalyses oxidative deamination of d-amino acids. This study aimed to explore the association between increased cerebral DAAO expression or enzymic activity and the development of cerebral ischaemia. EXPERIMENTAL APPROACH: A mouse model of transient (90 min) middle cerebral artery occlusion (MCAO) was established, and western blotting, enzymic activity assay, and fluorescent immunostaining techniques were used. KEY RESULTS: The expression and enzymic activity of DAAO increased over time in the cortical peri-infarct area of the mice subjected to transient MCAO. The DAAO was specifically expressed in astrocytes, and its double immunostaining with the astrocytic intracellular marker, glial fibrillary acidic protein, in the cortical peri-infarct area was up-regulated following ischaemic insult, with peak increase on Day 5 after MCAO. Single intravenous injection of the specific and potent DAAO inhibitor Compound SUN reduced the cerebral DAAO enzymic activity and attenuated neuronal infarction and neurobehavioural deficits with optimal improvement apparent immediately after the MCAO procedure. The neuroprotective effect was dose dependent, with ED50 values of 3.9-4.5 mg·kg-1 . Intracerebroventricular injection of the DAAO gene silencer siRNA/DAAO significantly reduced cerebral DAAO expression and attenuated MCAO-induced neuronal infarction and behavioural deficits. CONCLUSIONS AND IMPLICATIONS: Our results, for the first time, demonstrated that increased cerebral astrocytic DAAO expression and enzymic activity were causally associated with the development of neuronal destruction following ischaemic insults, suggesting that targeting cerebral DAAO could be a potential approach for treatment of neurological conditions following cerebral ischaemia.