Chemical Compositions of Essential Oil Extracted from Eight Thyme Species and Potential Biological Functions.

Thymus is an herbaceous perennial or subshrub of the Lamiaceae family and is widely distributed worldwide. Essential oils extracted from thymus have attracted much attention, owing to their potential biological functions. Here, we evaluated the chemical compositions of eight thyme essential oils (TEOs) using gas chromatography mass spectrometry and assessed their antioxidant activity. The results showed that (1) the main components in eight TEOs were monoterpene hydrocarbons and oxygenated monoterpenes (84.26-92.84%), and the chemical compositions of the TEOs were affected by the specie factor; (2) eight TEOs could be divided into three groups (thymol-, geraniol-, and nerol acetate-types), and thymol was the main type; (3) eight TEOs had some common compounds, such as thymol and p-cymene, which were the main components in seven TEOs; (4) eight TEOs had antioxidant activity, and Thymus pulegioides, Thymus thracicus, and Thymus serpyllum EOs had stronger antioxidant activity than vitamin E (0.07-0.27 fold) at a concentration of 1 mg/mL, while Thymus quinquecostatus and Thymus longicaulis EOs had relatively weak antioxidant activity. In addition, three chemical type standards were used to evaluate potential roles in antibacterial and tumor therapy. The results showed that thymol had strong antibacterial activity against the growth of Escherichia coli and Staphylococcus aureus, and antimigratory activity for A549 cell. Overall, our results can provide a theoretical basis for further exploring the function of natural products from thyme essential oils.

Automated classification of multiple ophthalmic diseases using ultrasound images by deep learning.

BACKGROUND: Ultrasound imaging is suitable for detecting and diagnosing ophthalmic abnormalities. However, a shortage of experienced sonographers and ophthalmologists remains a problem. This study aims to develop a multibranch transformer network (MBT-Net) for the automated classification of multiple ophthalmic diseases using B-mode ultrasound images. METHODS: Ultrasound images with six clinically confirmed categories, including normal, retinal detachment, vitreous haemorrhage, intraocular tumour, posterior scleral staphyloma and other abnormalities, were used to develop and evaluate the MBT-Net. Images were derived from five different ultrasonic devices operated by different sonographers and divided into training set, validation set, internal testing set and temporal external testing set. Two senior ophthalmologists and two junior ophthalmologists were recruited to compare the model's performance. RESULTS: A total of 10 184 ultrasound images were collected. The MBT-Net got an accuracy of 87.80% (95% CI 86.26% to 89.18%) in the internal testing set, which was significantly higher than junior ophthalmologists (95% CI 67.37% to 79.16%; both p<0.05) and lower than senior ophthalmologists (95% CI 89.45% to 92.61%; both p<0.05). The micro-average area under the curve of the six-category classification was 0.98. With reference to comprehensive clinical diagnosis, the measurements of agreement were almost perfect in the MBT-Net (kappa=0.85, p<0.05). There was no significant difference in the accuracy of the MBT-Net across five ultrasonic devices (p=0.27). The MBT-Net got an accuracy of 82.21% (95% CI 78.45% to 85.44%) in the temporal external testing set. CONCLUSIONS: The MBT-Net showed high accuracy for screening and diagnosing multiple ophthalmic diseases using only ultrasound images across mutioperators and mutidevices.

Total extract of Abelmoschus manihot L. alleviates uric acid-induced renal tubular epithelial injury via inhibition of caspase-8/caspase-3/NLRP3/GSDME signaling.

Purpose: The incidence of uric acid (UA)-induced kidney injury is increasing owing to the high incidence of hyperuricemia in recent years. The flower of Abelmoschus manihot (Linneus) Medik is a traditional Chinese medicinal herb widely used in the treatment of some kidney diseases. In our previous study, we reported that the total extract of A. manihot L. flower (TEA) attenuated adriamycin-induced renal tubular cell injury. In this study, we aimed to evaluate the role of TEA in UA-induced tubular cell injury. Methods: Normal rat proximal epithelial NRK-52E cells were incubated with UA to mimic hyperuricemia conditions. The role of TEA in the renal tubular cells was also assessed. The cellular morphology was observed using phase-contrast microscopy, and cell viability was analyzed using the Cell Counting kit-8. Living and dead cells were stained using a Calcein-AM/PI double stain kit. The release of lactate dehydrogenase (LDH) was analyzed by LDH cytotoxicity Assay Kit. The expression of target proteins was analyzed using western blot analysis. Results: UA triggered NRK-52E cell injury, as evidenced by morphological changes, detachment of cells from the bottom, cell swelling, large bubbles blowing from cell membrane and loss of cell viability. UA increased release of LDH. UA induced the expression of p-ERK1/2 and the subsequent activation of caspase-8, caspase-3, and NLRP3 inflammasomes. Pyroptosis was elicited by UA after gasdermin E N-terminal (GSDME-NT) was cleaved from gasdermin E (GSDME). Z-DEVD-FMK, a caspase-3 inhibitor, suppressed the expression of both NLRP3 and GSDME-NT, but not that of caspase-8. INF39, an NLRP3 inhibitor, altered the expression of GSDME-NT expression, but not that caspase-3 and caspase-8. TEA alleviated UA-induced cell injury by suppressing ERK1/2/caspase-8/caspase-3/NLRP3/GSDME signaling. Conclusion: GSDME-mediated pyroptosis was involved in UA-induced renal tubular cell injury. This is the first study to report that TEA protects renal tubular epithelial cells against UA by inhibiting the ERK/1/2/caspase-8/caspase-3/NLRP3/GSDME pathway.

The association between the serum uric acid to creatinine ratio and all-cause mortality in elderly hemodialysis patients.

BACKGROUND: Elderly hemodialysis patients have a higher rate of mortality than nonelderly hemodialysis patients. Recent studies shown that the serum uric acid to creatinine ratio (SUA/Scr) was associated with all-cause mortality in general adults. The purpose of the present study was to investigate the association between the SUA/Scr and all-cause and cardiovascular disease mortality among elderly hemodialysis patients. METHODS: A total of 222 patients (≥ 60 years) who received hemodialysis more than 8 h per week at Taizhou Second People's Hospital for at least 3 months were enrolled in the present study from January 2015 to December 2019. Clinical characteristics including age, sex and height et. al, were obtained from the hemodialysis database. The laboratory data, including albumin (ALB), total cholesterol (TC), serum uric acid (SUA), serum creatinine (Scr) and so on, were collected before hemodialysis and analyzed by automatic biochemical analyzer. Survival information was recorded during the follow-up period. Multiple Cox regression was carried out to analyze the association between SUA/Scr and all-cause mortality. The survival rate of each group was calculated by the Kaplan-Meier method, and the ratio of survival curves was analyzed by the log-rank test. The contribution of SUA/Scr for predicting all-cause mortality risk was evaluated by net reclassification improvement (NRI). RESULTS: During the 19-month observation period, 78 patients died. Individuals in the nonsurviving group had significantly older ages (P < 0.001), body mass index (BMI) (P = 0.004), serum creatinine (P = 0.005) and prealbumin (P = 0.006) than surviving patients. After adjusting for age, sex, BMI, prealbumin, dialysis vintage, dialysis frequency, single-pool Kt/V (spKt/V), DM, hypertension and comorbidities, a higher ratio of SUA/Scr was independently associated with a higher risk of all-cause mortality (HR: 1.292; 95% CI: 1.013-1.648; P = 0.039). The predict value on all-cause mortality of SUA/Scr was superior to SUA (additive NRI = 0.214, P = 0.015) and Scr (additive NRI = 0.476, P < 0.001) among elderly hemodialysis patients. CONCLUSION: The serum uric acid to creatinine ratio is strongly associated with all-cause mortality in elderly hemodialysis patients which is more predictive than SUA or Scr alone.

MAC-ResNet: Knowledge Distillation Based Lightweight Multiscale-Attention-Crop-ResNet for Eyelid Tumors Detection and Classification.

Eyelid tumors are tumors that occur in the eye and its appendages, affecting vision and appearance, causing blindness and disability, and some having a high lethality rate. Pathological images of eyelid tumors are characterized by large pixels, multiple scales, and similar features. Solving the problem of difficult and time-consuming fine-grained classification of pathological images is important to improve the efficiency and quality of pathological diagnosis. The morphology of Basal Cell Carcinoma (BCC), Meibomian Gland Carcinoma (MGC), and Cutaneous Melanoma (CM) in eyelid tumors are very similar, and it is easy to be misdiagnosed among each category. In addition, the diseased area, which is decisive for the diagnosis of the disease, usually occupies only a relatively minor portion of the entire pathology section, and screening the area of interest is a tedious and time-consuming task. In this paper, deep learning techniques to investigate the pathological images of eyelid tumors. Inspired by the knowledge distillation process, we propose the Multiscale-Attention-Crop-ResNet (MAC-ResNet) network model to achieve the automatic classification of three malignant tumors and the automatic localization of whole slide imaging (WSI) lesion regions using U-Net. The final accuracy rates of the three classification problems of eyelid tumors on MAC-ResNet were 96.8%, 94.6%, and 90.8%, respectively.

Sublytic C5b-9 triggers glomerular mesangial cell proliferation via enhancing FGF1 and PDGFα gene transcription mediated by GCN5-dependent SOX9 acetylation in rat Thy-1 nephritis.

Rat Thy-1 nephritis (Thy-1N) is an animal model of human mesangioproliferative glomerulonephritis (MsPGN), accompanied by glomerular mesangial cell (GMC) proliferation and extracellular matrix (ECM) deposition. Although sublytic C5b-9 formed on GMC membrane could induce cell proliferation, the mechanism is still unclear. In this study, we first demonstrated that the level of SRY related HMG-BOX gene 9 (SOX9), general control nonderepressible 5 (GCN5), fibroblast growth factor 1 (FGF1) and platelet-derived growth factor α (PDGFα) was all elevated both in the renal tissues of Thy-1N rats (in vivo) and in the GMCs (in vitro) with sublytic C5b-9 stimulation. Then, we not only discovered that sublytic C5b-9 caused GMC proliferation through increasing SOX9, GCN5, FGF1 and PDGFα expression, but also proved that SOX9 and GCN5 formed a complex and combined with FGF1 and PDGFα promoters, leading to FGF1 and PDGFα gene transcription. More importantly, GCN5 could mediate SOX9 acetylation at lysine 62 (K62) to enhance SOX9 binding to FGF1 or PDGFα promoter and promote FGF1 or PDGFα synthesis and GMC proliferation. Besides, the experiments in vivo also showed that FGF1 and PDGFα expression, GMC proliferation and urinary protein secretion in Thy-1N rats were greatly reduced by silencing renal SOX9, GCN5, FGF1 or PDGFα gene. Furthermore, the renal tissues of MsPGN patients also exhibited positive expression of these genes mentioned above. Collectively, our findings indicate that GCN5, SOX9 and FGF1/PDGFα can form an axis and play an essential role in sublytic C5b-9-triggered GMC proliferation, which might provide a novel insight into the pathogenesis of Thy-1N and MsPGN.

Sublytic C5b-9 induces glomerular mesangial cell proliferation via ERK1/2-dependent SOX9 phosphorylation and acetylation by enhancing Cyclin D1 in rat Thy-1 nephritis.

Glomerular mesangial cell (GMC) proliferation is a histopathological alteration in human mesangioproliferative glomerulonephritis (MsPGN) or in animal models of MsPGN, e.g., the rat Thy-1 nephritis (Thy-1N) model. Although sublytic C5b-9 assembly on the GMC membrane can trigger cell proliferation, the mechanisms are still undefined. We found that sublytic C5b-9-induced rat GMC proliferation was driven by extracellular signal-regulated kinase 1/2 (ERK1/2), sry-related HMG-box 9 (SOX9), and Cyclin D1. Here, ERK1/2 phosphorylation was a result of the calcium influx-PKC-α-Raf-MEK1/2 axis activated by sublytic C5b-9, and Cyclin D1 gene transcription was enhanced by ERK1/2-dependent SOX9 binding to the Cyclin D1 promoter (-582 to -238 nt). In addition, ERK1/2 not only interacted with SOX9 in the cell nucleus to mediate its phosphorylation at serine residues 64 (a new site identified by mass spectrometry) and 181 (a known site), but also indirectly induced SOX9 acetylation by elevating the expression of general control non-repressed protein 5 (GCN5), which together resulted in Cyclin D1 synthesis and GMC proliferation. Moreover, our in vivo experiments confirmed that silencing these genes ameliorated the lesions of Thy-1N rats and reduced SOX9 phosphorylation, acetylation and Cyclin D1 expression. Furthermore, the renal tissue sections of MsPGN patients also showed higher phosphorylation or expression of ERK1/2, SOX9, and Cyclin D1. In summary, these findings suggest that sublytic C5b-9-induced GMC proliferation in rat Thy-1N requires SOX9 phosphorylation and acetylation via enhanced Cyclin D1 gene transcription, which may provide a new insight into human MsPGN pathogenesis.

An accurate interactive segmentation and volume calculation of orbital soft tissue for orbital reconstruction after enucleation.

BACKGROUND: Accurate measurement and reconstruction of orbital soft tissue is important to diagnosis and treatment of orbital diseases. This study applied an interactive graph cut method to orbital soft tissue precise segmentation and calculation in computerized tomography (CT) images, and to estimate its application in orbital reconstruction. METHODS: The interactive graph cut method was introduced to segment extraocular muscle and intraorbital fat in CT images. Intra- and inter-observer variability of tissue volume measured by graph cut segmentation was validated. Accuracy and reliability of the method was accessed by comparing with manual delineation and commercial medical image software. Intraorbital structure of 10 patients after enucleation surgery was reconstructed based on graph cut segmentation and soft tissue volume were compared within two different surgical techniques. RESULTS: Both muscle and fat tissue segmentation results of graph cut method showed good consistency with ground truth in phantom data. There were no significant differences in muscle calculations between observers or segmental methods (p > 0.05). Graph cut results of fat tissue had coincidental variable trend with ground truth which could identify 0.1cm3 variation. The mean performance time of graph cut segmentation was significantly shorter than manual delineation and commercial software (p < 0.001). Jaccard similarity and Dice coefficient of graph cut method were 0.767 ± 0.045 and 0.836 ± 0.032 for human normal extraocular muscle segmentation. The measurements of fat tissue were significantly better in graph cut than those in commercial software (p < 0.05). Orbital soft tissue volume was decreased in post-enucleation orbit than that in normal orbit (p < 0.05). CONCLUSION: The graph cut method was validated to have good accuracy, reliability and efficiency in orbit soft tissue segmentation. It could discern minor volume changes of soft tissue. The interactive segmenting technique would be a valuable tool for dynamic analysis and prediction of therapeutic effect and orbital reconstruction.

Simultaneous enhancement of vascularization and contact-active antibacterial activity in diopside-based ceramic orbital implants.

Rapid vascularization and long-term antibacterial property are desirable characteristics of the next-generation implants in orbital reconstruction. In this study, the new diopside-based orbital implants were developed by direct ink writing of diopside (CaMgSi2O6; DIO) and low-melt bioactive glass (BG)-assisted sintering approaches. The mechanical tests showed that the addition 5% or 10% BG could readily enhance the compressive strength of the DIO porous bioceramics after sintering at 1150 °C. The Tris buffer immersion test in vitro indicated that the porous bioceramics exhibited appreciable mechanical stability and very limited mass loss (<3.5%) after 8 weeks. The DIO/10BG porous bioceramic sintered at 1150 °C or 1250 °C could promote appreciable angiogenesis response at the early stage (2-6 weeks) of implantation in the rabbit panniculus carnosus muscle models in vivo. It is interesting that the steam autoclaved bioceramics exhibited outstanding contact-active inhibition against Staphylococcus aureus and Pseudomonas aeruginosa, but as-sintered bioceramics showed no antibacterial effect. It is reasonable to consider that our strategy paves the way toward a simple and effective approach to fabricate the multifunctional tailormade implants for orbital implantation, thus accelerating the clinical translation of biomaterials research.

Sublytic C5b-9 induces proliferation of glomerular mesangial cells via ERK5/MZF1/RGC-32 axis activated by FBXO28-TRAF6 complex.

Mesangioproliferative glomerulonephritis (MsPGN) is characterized by the proliferation of glomerular mesangial cells (GMCs) and accumulation of extracellular matrix (ECM), followed by glomerulosclerosis and renal failure of patients. Although our previous studies have demonstrated that sublytic C5b-9 complex formed on the GMC membrane could trigger GMC proliferation and ECM expansion of rat Thy-1 nephritis (Thy-1N) as an animal model of MsPGN, their mechanisms are still not fully elucidated. In the present studies, we found that the levels of response gene to complement 32 (RGC-32), myeloid zinc finger 1 (MZF1), phosphorylated extracellular signal-regulated kinase 5 (phosphorylated ERK5, p-ERK5), F-box only protein 28 (FBXO28) and TNF receptor-associated factor 6 (TRAF6) were all markedly up-regulated both in the renal tissues of rats with Thy-1N (in vivo) and in the GMCs upon sublytic C5b-9 stimulation (in vitro). Further in vitro experiments revealed that up-regulated FBXO28 and TRAF6 could form protein complex binding to ERK5 and enhance ERK5 K63-ubiquitination and subsequent phosphorylation. Subsequently, ERK5 activation contributed to MZF1 expression and MZF1-dependent RGC-32 up-regulation, finally resulting in GMC proliferative response. Furthermore, the MZF1-binding element within RGC-32 promoter and the functions of FBXO28 domains were identified. Additionally, knockdown of renal FBXO28, TRAF6, ERK5, MZF1 and RGC-32 genes respectively markedly reduced GMC proliferation and ECM production in Thy-1N rats. Together, these findings indicate that sublytic C5b-9 induces GMC proliferative changes in rat Thy-1N through ERK5/MZF1/RGC-32 axis activated by the FBXO28-TRAF6 complex, which might provide a new insight into MsPGN pathogenesis.

Synergistic Effect of Copper-Containing Mesoporous Bioactive Glass Coating on Stimulating Vascularization of Porous Hydroxyapatite Orbital Implants in Rabbits.

Rapid and complete vascularization of the porous orbital implants after enucleation is vital for reducing postoperative complications, such as exposure and infection. In this study, the porous hydroxyapatite (pHA) scaffolds modified with the mesoporous bioactive glass with and without adding copper (MBG, Cu-MBG) were prepared by sol-gel technique and followed by evaporation induced self-assembly (EISA) process. The macro-/microstructure analysis of the modified scaffolds (i.e., MBG/pHA, Cu-MBG/pHA) showed highly interconnected macropores, and unique mesoporous structures in the pore wall with higher surface area. A primary angiogenic test in a panniculus carnosus muscle model in rabbit indicated that the Cu-MBG coating was significantly beneficial for the vascularization in the porous architecture of the scaffolds in the early stage. These results suggest that the Cu-MBG modification provides a simple and effective strategy to endow favorable biological performances with pHA implants and greater potential for future application in orbit reconstruction.

Global Patterns in Health Burden of Uncorrected Refractive Error.

Purpose: To evaluate the global patterns in health burden of uncorrected refractive error (URE) by year, age and sex, region, and socioeconomic status, using disability-adjusted life years (DALYs). Methods: Global, regional, or national DALY numbers, crude DALY rates, and age-standardized DALY rates caused by URE, by year, or age and sex, were obtained from the Global Burden of Disease Study 2013. Human development index (HDI) in 2013 as a national socioeconomic indicator was obtained from the Human Development Report. Kruskal-Wallis test and linear regression were performed to explore the association between age-standardized DALY rates and HDI. Results: From 1990 to 2013, global DALY numbers caused by URE rose by 43.8%, crude rates remained relatively constant, and age-standardized rates fell by 8.8%. Global DALY rates increased with age, and older females had higher DALY numbers and rates than males of the same age. Age-standardized DALY rates in Eastern Mediterranean, South-East Asia, and Africa were higher than that at a global level. Multiple comparisons indicated higher age-standardized DALY rates in lower HDI countries. Age-standardized DALY rates were inversely related to HDI (standardized ß = -0.616, P < 0.001). Conclusions: The global health of URE is improving but crude DALY rates are keeping constant, implying that health progress does not mean fewer demands of refractive services. Worldwide, older age, female sex, and lower socioeconomic status are associated with higher URE burden. The findings of this study may raise public awareness of the global URE burden and are important for health policy making.

16-Channel Organic Electrochemical Transistor Array for In Vitro Conduction Mapping of Cardiac Action Potential.

16-Channel organic electrochemical transistor arrays (OECTs) are developed for mapping the propagation and studying the characteristics of action potentials of primary cardiomyocytes. The physiological activities of a rat cardiomyocyte monolayer during a long-term culturing is revealed by this biocompatible, low-cost, and high transconductance organic electronic device. OECT has great potential to be used in cardiac and neuronal drug screening.

Rational Design and Fabrication of Porous Calcium-Magnesium Silicate Constructs That Enhance Angiogenesis and Improve Orbital Implantation.

Tissue integration of orbital implants, following orbital enucleation treatment, represents a challenge for rapid fibrovascularization, long-time stability, anti-infection, and even induction of vascule regeneration. The objective of this study was to develop porous calcium-magnesium silicate materials, with good stability, bioactivity, and antibacterial potential as new orbital fillers. Three-dimensional (3D) diopside scaffolds (low dissolvability) were fabricated by direct ceramic ink writing assembly and then followed by one-step sintering at 1150 °C for 3 h. The pore wall of the scaffold was modified by another calcium-magnesium silicate, such as bredigite or akermanite, which dissolves quickly but shows greater angiogenic potential. These two Ca-Mg-silicates can be coated onto the pore strut, and the coating layers were observed to slowly dissolve in Tris buffer. The vascularization-favorable Cu ions, which had been doped into the bredigite or akermanite coating, could also be measured in the immersion medium. A primary angiogenic test in a panniculus carnosus muscle model in rabbit indicated that the Cu-doped bredigite and akermanite coatings were significantly beneficial for the neovascularization in the early stages. These results suggest that the diopside-based porous materials modified with functional coatings hold great potential for application in orbital reconstruction.

Rigid and flexible organic electrochemical transistor arrays for monitoring action potentials from electrogenic cells.

Rigid and flexible organic electrochemical transistor arrays are successfully implemented for monitoring cardiac action potentials. Excellent signal to noise ratios are achieved with values routinely larger than 4. These devices are promising to be used in both conventional and emerging areas.

Engineering a freestanding biomimetic cardiac patch using biodegradable poly(lactic-co-glycolic acid) (PLGA) and human embryonic stem cell-derived ventricular cardiomyocytes (hESC-VCMs).

Microgrooved thin PLGA film (≈30 µm) is successfully fabricated on a Teflon mold, which could be readily peeled off and is used for the construction of a biomimetic cardiac patch. The contraction of it is studied with optical mapping on transmembrane action potential. Our results suggest that steady-state contraction could be easily established on it under regular electrical stimuli. Besides, the biomimetic cardiac patch recapitulates the anisotropic electrophysiological feature of native cardiac tissue and is much more refractory to premature stimuli than the random one constructed with non-grooved PLGA film, as proved by the reduced incidence of arrhythmia. Considering the good biocompatibility of PLGA as demonstrated in our study and the biodegradability of it, our biomimetic cardiac patch may find applications in the treatment of myocardial infarction. Moreover, the Teflon mold could be applied in the fabrication of various scaffolds with fine features for other tissues.

The Ras GTPase-activating-like protein IQGAP1 is downregulated in human diabetic nephropathy and associated with ERK1/2 pathway activation.

Podocyte injury may contribute to the pathogenesis of diabetic nephropathy (DN), but the underlying mechanism of hyperglycemia induced podocyte damage is not fully understood. The Ras GTPase-activating-like protein IQGAP1 is associated to the slit diaphragm proteins and the actin cytoskeleton in podocyte. Here, we studied IQGAP1 expression alterations in human DN biopsies and extracellular signal-regulated kinase (ERK)-dependent pathways of IQGAP1 expression in podocyte under high glucose (HG) media. In vivo, analysis of renal biopsies from patients with DN revealed a significant reduction in IQGAP1 expression compared to controls. In vitro, IQGAP1 mRNA and protein expression were observed to decline under HG media at 48 h. But phosphorylation of ERK1/2 was activated under HG media at 24 h and 48 h. However, HG-induced downregulation of IQGAP1 protein was attenuated by specific ERK1/2 activation inhibitor PD98059. Taken together, these results highlight the importance of IQGAP1 in DN, and suggest that IQGAP1 expression in podocyte under HG media is modulated by the ERK1/2 pathway, which may lead to the future development of therapies targeting IQGAP1 dysfunction in podocytes in DN.

Organic electrochemical transistor array for recording transepithelial ion transport of human airway epithelial cells.

An organic electrochemical transistor array is integrated with human airway epithelial cells. This integration provides a novel method to couple transepithelial ion transport with electrical current. Activation and inhibition of transepithelial ion transport are readily detected with excellent time resolution. The organic electrochemical transistor array serves as a promising platform for physiological studies and drug testing.

Staining-free gel electrophoresis-based multiplex enzyme assay using DNA and peptide dual-functionalized gold nanoparticles.

We report a simple staining-free gel electrophoresis method to simultaneously probe protease and nuclease. Utilizing gold nanoparticles (Au-NPs) dual-functionalized with DNA and peptide, the presence and concentration of nuclease and protease are determined concurrently from the relative position and intensity of the bands in the staining-free gel electrophoresis. The use of Au-NPs eliminates the need for staining processes and enables naked eye detection, while a mononucleotide-mediated approach facilitates the synthesis of DNA/peptide conjugated Au-NPs and simplifies the operation procedures. Multiplex detection and quantification of DNase I and trypsin are successfully demonstrated.

[Protective effects of luteolin on neurons against oxygen-glucose deprivation/reperfusion injury via improving Na+/K+ -ATPase activity].

OBJECTIVE: Luteolin, a flavone, has considerable neuroprotective effects by its anti-oxidative mechanism. However, it is still unclear whether luteolin can protect neurons against oxygen-glucose deprivation/reperfusion (OGD/R) induced injury. METHOD: After 2 hours oxygen-glucose deprivation and 24 hours reperfusion treatment in primary cultured hippocampal neurons, the neuron viability, survival rate and apoptosis rate were evaluated by MTT assay, lactate dehydrogenase (LDH) leakage assay and Hoechst staining, respectively. The activity of Na+/K+ -ATPase was examined in cultured neurons or in the hippocampus of SD rats treated by 10 minutes global cerebral ischemia and followed 24 hours reperfusion. RESULT: Treatment by OGD/R markedly reduced neuronal viability, increased LDH leakage rate and increased apoptosis rate. Application of luteolin (10-100 micromol x L(-1)) during OGD inhibited OGD/R induced neuron injury and apoptosis in a dose-dependent manner. Compared to the control group or OGP/R-treated neurons, the activity of Na+/K+ -ATPase was significantly suppressed in global ischemia/reperfusion group or OGD/R-treated neurons. Application of luteolin during ischemia or OGD preserved the Na+/K+ -ATPase activity. Furthermore, inhibition of Na+/K+ -ATPase with ouabain attenuated the protective effect afforded by luteolin. CONCLUSION: The data provide the evidence that luteolin has neuroprotective effect against OGD/R induced injury and the protective effect may be associated with its ability to improve Na+/K+ -ATPase activity after OGD/R.