Engineering Cardiac Tissue for Advanced Heart-On-A-Chip Platforms.

Cardiovascular disease is a major cause of mortality worldwide, and current preclinical models including traditional animal models and 2D cell culture models have limitations in replicating human native heart physiology and response to drugs. Heart-on-a-chip (HoC) technology offers a promising solution by combining the advantages of cardiac tissue engineering and microfluidics to create in vitro 3D cardiac models, which can mimic key aspects of human microphysiological systems and provide controllable microenvironments. Herein, recent advances in HoC technologies are introduced, including engineered cardiac microtissue construction in vitro, microfluidic chip fabrication, microenvironmental stimulation, and real-time feedback systems. The development of cardiac tissue engineering methods is focused for 3D microtissue preparation, advanced strategies for HoC fabrication, and current applications of these platforms. Major challenges in HoC fabrication are discussed and the perspective on the potential for these platforms is provided to advance research and clinical applications.

Application of Persimmon Pectin with Promising Emulsification Properties as an Acidified Milk Drinks Stabilizer.

The present study aimed to evaluate the capability of persimmon pectin (PP) as a stabilizer for acid milk drinks (AMDs) compared with commercial high-methoxyl pectin (HMP) and sugar beet pectin (SBP). The effectiveness of pectin stabilizers was assessed by analyzing particle size, micromorphology, zeta potential, sedimentation fraction, storage, and physical stability. Results of CLSM images and particle size measurements showed that PP-stabilized AMDs had smaller droplet sizes and more uniform distributions, indicating better stabilization potential compared with the HMP- and SBP-stabilized AMDs. Zeta potential measurements revealed that the addition of PP significantly increased the electrostatic repulsion between particles and prevented aggregation. Moreover, based on the results of Turbiscan and storage stability determination, PP exhibited better physical and storage stability compared with HMP and SBP. The combination of steric repulsion and electrostatic repulsion mechanisms exerted a stabilizing effect on the AMDs prepared from PP. Overall, these findings suggest that PP has promising potential as an AMD stabilizer in the food and beverage industry.

A Deep Learning Method for Breast Cancer Classification in the Pathology Images.

OBJECTIVE: Breast cancer is the most common female cancer in the world, and it poses a huge threat to women's health. There is currently promising research concerning its early diagnosis using deep learning methodologies. However, some commonly used Convolutional Neural Network (CNN) and their variations, such as AlexNet, VGGNet, GoogleNet and so on, are prone to overfitting in breast cancer classification, due to both small-scale breast pathology image datasets and overconfident softmax-cross-entropy loss. To alleviate the overfitting issue for better classification accuracy, we propose a novel framework for breast pathology classification, called the AlexNet-BC model. The model is pre-trained using the ImageNet dataset and fine-tuned using an augmented dataset. We also devise an improved cross-entropy loss function to penalize overconfident low-entropy output distributions and make the predictions suitable for uniform distributions. The proposed approach is then validated through a series of comparative experiments on BreaKHis, IDC and UCSB datasets. The experimental results show that the proposed method outperforms the state-of-the-art methods at different magnifications. Its strong robustness and generalization capabilities make it suitable for histopathology clinical computer-aided diagnosis systems.

Achieving Ammonium Removal Through Anammox-Derived Feammox With Low Demand of Fe(III).

Feammox-based nitrogen removal technology can reduce energy consumption by aeration and emission of carbon dioxide. However, the huge theoretical demand for Fe(III) becomes a challenge for the further development of Feammox. This study investigated an anammox-derived Feammox process with an intermittent dosage of Fe2O3 and proposed a novel approach to reduce the Fe(III) consumption. The results showed that anammox genera Candidatus Brocadia and Candidatus Kuenenia in the seed anammox sludge significantly decreased after cultivation. The formation of N2 was the dominating pathway in Feammox while that of nitrite and nitrate could be neglected. Batch tests showed that specific Feammox activity of ammonium oxidation was 1.14-9.98 mg N/(g VSS·d). The maximum removal efficiency of ammonium reached 52.3% in the bioreactor with a low dosage of Fe(III) which was only 5.8% of the theoretical demand in Feammox. The removal of ammonium was mainly achieved through Feammox, while partial nitrification/anammox also played a role due to the non-power and unintentional oxygen leakage. The super-low oxygen also responded to the low demand of Fe(III) in the bioreactor because it could trigger the cycle of Fe(III)/Fe(II) by coupling Feammox and chemical oxidation of Fe(II) to Fe(III). Therefore, anammox-derived Feammox can achieve the removal of ammonium with low Fe(III) demand at super-low oxygen.

Neuroprotection of retinal cells by Caffeic Acid Phenylethyl Ester（CAPE) is mediated by mitochondrial uncoupling protein UCP2.

Oxidative stress due to mitochondrial produced reactive oxygen species is a major cause of damage seen in many retinal degenerative diseases. Caffeic acid phenylethyl ester （CAPE） is protective agent in multiple tissues and is reported to have anti-oxidant properties. Systemically applied CAPE protected retinal ganglion cells from ischemic injury induced by increased intraocular pressure. CAPE provided complete protection for ARPE19 retinal pigment epithelial cells against tert-butyl hydrogen peroxide and reduced both basal and LPS-stimulated ROS production. The major effect of CAPE was mediated by the mitochondrial uncoupling protein UCP2 since both pharmacological inhibition of UCP2 and siRNA-induced knockdown removed the ability of CAPE to block ROS production. Based on common structural features, CAPE may be acting as a mimetic of the natural UCP2 homeostatic regulator 4-hydroxy-2-nonenal. CAPE may provide a valuable tool to treat oxidative stress-related damage in retinal and other degenerative diseases.

Hyperoside Alleviates High Glucose-Induced Proliferation of Mesangial Cells through the Inhibition of the ERK/CREB/miRNA-34a Signaling Pathway.

PURPOSE: Hyperoside, a flavonoid isolated from conventional medicinal herbs, has been demonstrated to exert a significant protective effect in diabetic nephropathy. This study aimed to determine the underlying mechanisms, by which hyperoside inhibits high glucose-(HG-) induced proliferation in mouse renal mesangial cells. METHODS: Mouse glomerular mesangial cells line (SV40-MES13) was used to study the inhibitory effect of hyperoside on cell proliferation induced by 30 mM glucose, which was used to simulate a diabetic condition. Viable cell count was assessed using the Cell Counting Kit-8 and by the 5-ethynyl-20-deoxyuridine incorporation assay. The underlying mechanism involving miRNA-34a was further investigated by quantitative RT-PCR and transfection with miRNA-34a agomir. The phosphorylation levels of extracellular signal-regulated kinases (ERKs) and cAMP-response element-binding protein (CREB) were measured by Western blotting. The binding region and the critical binding sites of CREB in the miRNA-34a promoter were investigated by the chromatin immunoprecipitation assay and luciferase reporter assay, respectively. RESULTS: We found that hyperoside could significantly decrease HG-induced proliferation of SV40-MES13 cells in a dose-dependent manner, without causing obvious cell death. In addition, hyperoside inhibited the activation of ERK pathway and phosphorylation of its downstream transcriptional factor CREB, as well as the miRNA-34a expression. We further confirmed that CREB-mediated regulation of miRNA-34a is dependent on the direct binding to specific sites in the promoter region of miRNA-34a. CONCLUSION: Our cumulative results suggested that hyperoside inhibits the proliferation of SV40-MES13 cells through the suppression of the ERK/CREB/miRNA-34a signaling pathway, which provides new insight to the current investigation on therapeutic strategies for diabetic nephropathy.

Effects of Crude ß-Glucosidases from Issatchenkia terricola, Pichia kudriavzevii, Metschnikowia pulcherrima on the Flavor Complexity and Characteristics of Wines.

To investigate the effects of crude ß-glucosidases from Issatchenkia terricola SLY-4 (SLY-4E), Pichia kudriavzevii F2-24 (F2-24E), and Metschnikowia pulcherrima HX-13 (HX-13E) on flavor complexity and characteristics of wines, grape juice was fermented by Saccharomyces cerevisiae with the addition of SLY-4E, F2-24E and HX-13E, respectively. The growth and sugar consumption kinetics of S. cerevisiae, the physicochemical characteristics, the volatile compounds, and the sensory dimensions of wines were analyzed. Results showed that adding SLY-4E, F2-24E, and HX-13E into must had no negative effect on the fermentation and physicochemical characteristics of wines, but increased the content of terpenes, esters, and fatty acids, while decreased the C6 compound content. Each wine had its typical volatile compound profiles. Adding SLY-4E or F2-24E into must could significantly improve the flavor complexity and characteristics of wines. These results would provide not only an approach to improve flavor complexity and characteristics of wines, but also references for application of ß-glucosidases from other sources.

The Sensory Quality Improvement of Citrus Wine through Co-Fermentations with Selected Non-Saccharomyces Yeast Strains and Saccharomyces cerevisiae.

Co-fermentation of selected non-Saccharomyces yeast strain with Saccharomyces cerevisiae is regarded as a promising approach to improve the sensory quality of fruit wine. To evaluate the effects of co-fermentations between the selected non-Saccharomyces yeast strains (Hanseniaspora opuntiae, Hanseniaspora uvarum and Torulaspora delbrueckii) and S. cerevisiae on the sensory quality of citrus wine, the fermentation processes, the chemical compositions, and the sensory evaluations of citrus wines were analyzed. Compared with those of S. cerevisiae fermentation, co-fermentations produced high sensory qualities, and S. cerevisiae/H. opuntiae co-fermentation had the best sensory quality followed by Sc-Hu and Sc-Td co-fermentations. Additionally, all the co-fermentations had a lower amount of ethanol and total acidity, higher pH value, and higher content of volatile aroma compounds, especially the content of higher alcohol and ester compounds, than those of S. cerevisiae fermentation. Therefore, co-fermentations of the non-Saccharomyces yeast strains and S. cerevisiae could be employed to improve the sensory quality of citrus wines. These results would provide not only methods to improve the sensory quality of citrus wine, but also a valuable reference for the selection of non-Saccharomyces yeast strains for fruit wine fermentation.

Engineering Saccharomyces cerevisiae Coculture Platform for the Production of Flavonoids.

Flavonoids are valuable natural products widely used in human health and nutrition applications. Engineering microbial consortia to express complex flavonoid biosynthetic pathways is a promising approach for flavonoid production. In this study, the entire flavonoid biosynthetic pathway was split into two independent pathways, each of which was contained in separate Saccharomyces cerevisiae cells. The first cell type, sNAR5, which was genetically engineered to express the naringenin biosynthetic pathway, produced 144.1 mg/L naringenin. The second cell type was genetically modified with the heterologous naringenin-to-delphinidin pathway. A coculture produced a delphinidin titer, significantly higher than that produced in a monoculture of strain sDPD2, harboring the entire pathway. Furthermore, we successfully employed this coculture platform for the production of 3 flavonols and 2 anthocyanidins in flask-scale culture. This coculture platform paves the way for the development of an economical and efficient process for microbial flavonoid production.

6-Gingerol ameliorates sepsis-induced liver injury through the Nrf2 pathway.

Sepsis-induced liver injury is very common in intensive care units. Here, we investigated the effects of 6-gingerol on sepsis-induced liver injury and the role of the Nrf2 pathway in this process. 6-Gingerol is the principal ingredient of ginger that exerts anti-inflammatory and antioxidant effects. Using cecal ligation and puncture (CLP) to induce polymicrobial sepsis and related liver injury, we found that mice pre-treated with 6-Gingerol showed less incidences of severe liver inflammation and death than untreated CLP groups. 6-Gingerol administration also inhibited the expression of pyroptosis-related proteins, including NOD-like receptor protein 3 (NLRP3), IL-1ß, and caspase-1. Consistent with these findings, 6-gingerol reduced the effects of pyroptosis induced by lipopolysaccharide (LPS) and adenosine 5'-triphosphate (ATP) in RAW 264.7 cells, as evidenced by IL-1ß and caspase-1 protein levels in the supernatant and propidium iodide (PI) staining. 6-Gingerol was shown to activate the Nrf2 pathway in vivo and in vitro. Notably, Nrf2 siRNA transfection nullified the inhibitory effects of 6-gingerol on pyroptosis in vitro. In summary, these findings suggested that 6-gingerol alleviated sepsis-induced liver injury by inhibiting pyroptosis through the Nrf2 pathway.

Arrayed microfluidic chip for detection of circulating tumor cells and evaluation of drug potency.

Circulating tumor cells (CTCs) from peripheral blood of cancer patients are considered as one of the most promising pharmacodynamic (PD) biomarkers due to its non-invasive property in disease diagnosis and prognosis. However, the detection of extremely low number of CTCs in patient blood requires methods with high sensitivity and accuracy. We fabricated an arrayed geometrically enhanced mixing (GEM) chip with a "dislocation herringbone" layout based on cell immunoaffinity. By optimizing the injection and rinsing flow rate, an average cell capture rate of 87.02% and an average capture purity of 99.58% were achieved using the human lung adenocarcinoma cell lines H1975. In addition, we determined the specificity, precision, accuracy, and detection limit of our chip. The results demonstrated the chip was stable, accurate and reliable for the "liquid biopsy" of lung cancer cells using the peripheral blood of patients. Our chip can also be used to evaluate the potency of different drugs against tumor cells in parallel due to the presence of four independent microchannels.

(E)­phenethyl 3­(3,5­dihydroxy­4­isopropylphenyl) acrylate gel improves DNFB-induced allergic contact hypersensitivity via regulating the balance of Th1/Th2/Th17/Treg cell subsets.

(E)­phenethyl 3­(3,5­dihydroxy­4­isopropylphenyl) acrylate gels (THCA354) is a novel polyphenols acrylic acid derivative. To investigate the immunoregulatory mechanisms of THCA354, we established a mouse model of 2,4­dinitrofluorobenzene (DNFB)-induced allergic contact dermatitis (ACD). Responses of Th1, Th2, Th17 and regulatory T cells (Tregs) were determined by flow cytometry, reverse-transcriptase polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA). Our study found that topical application of THCA354 gel could inhibit ear swelling, reduce inflammatory cell infiltration, down-regulate Th1/Th17 responses and enhance Th2/Treg responses. These findings indicated that THCA354 gel exerted its immunotherapeutic effects by modulating the balance of Th1/Th2/Th17/Treg cell subsets, suggesting that THCA354 gel could be used as a promising drug candidate for intervention of ACD.

Selection of non-Saccharomyces yeasts for orange wine fermentation based on their enological traits and volatile compounds formation.

In order to select the non-Saccharomyces yeasts for orange wine fermentation, the enological traits and volatile compounds formation of ten non-Saccharomyces yeast strains were evaluated through physicochemical methods and solid-phase microextraction coupled to GC-MS, respectively. The results indicated that non-Saccharomyces yeast fermentation had lower maximum populations (7.8-8.0 Log cfu/mL), longer fermentation period (7-10 days), lower ethanol (4.13-7.79%), lower total acids (7.48-8.51 g/L) and higher volatile acids concentrations (0.08-0.23 g/L) when compared with those of Saccharomyces cerevisiae fermentation. Hanseniaspora uvarum, Hanseniaspora opuntiae, Hanseniaspora occidentalis, Pichia kudriavzevii and Torulaspora delbrueckii were selected as candidates for orange wine fermentation with higher volatile compounds concentration, odor active values and sensory evaluation scores. This study will provide a valuable selection method of non-Saccharomyces yeasts for orange wine fermentation, and an approach to improve the flavor of orange wine or other fruit wine.

Physicochemical and structural characteristics of the octenyl succinic ester of ginkgo starch.

The physicochemical and structural characteristics of octenyl succinate (OS)-starch prepared from ginkgo starch were investigated. The degree of substitution (DS) and reaction efficiency of 1.5 and 3.0% OSA-modified starch were 0.0065, 77.9% and 0.0172, 70.4%, respectively. OSA modification caused a reduction in glucose yield but no significant changes in the z-root mean square radius of gyration (Rz), even though the weight-average molar weight (Mw) and molecular density (ρ) were reduced. Two newabsorption bands (1725 and 1570cm-1) were observed for OS-starch. The heat of gelatinization (ΔH), onset (To), peak (Tp), and conclusion (Tc) temperatures decreased as the OSA level increased. OSA level did not significantly affect relative crystallinity (Xc), showing that OSA esterification occurred primarily in the amorphous domain. The rheological characteristics of OS-starch pastes indicated highly shear-thinning fluids. This study showed that changes in digestibility and rheological properties of OS-starch depended both on DS and starch structure.

The optimization of isoamylase processing conditions for the preparation of high-amylose ginkgo starch.

A high-amylose starch was prepared from ginkgo by hydrolysis using isoamylase and its structures (morphology and crystallinity) and physicochemical properties (swelling factor, water solubility and gelatinization) were determined. The experiments used response surface methodology to determine the optimum parameters for enzymatic hydrolysis: pH 5.0 at 52 °C for 170 min, using an enzyme dose greater than 100 IU/ml. The experimentally observed maximum yield of ginkgo amylose under these conditions was 74.74% and the blue value was 0.756. The high-amylose ginkgo starch showed an irregular surface and porous inner structure while the native starch granules were oval with a smooth surface. X-ray showed that the high-amylose starch displayed a V-type structure. Because of its high amylose content and different structural characteristics, high-amylose starch exhibited a higher gelatinization peak temperature (109.25 °C) and water solubility, and a lower crystallinity (19.13%), gelatinization enthalpy (63.83 J/g), and swelling power. The present study has indicated that high-amylose starch prepared using isoamylase has unique functional properties, which lays the foundation for the wider application of ginkgo starch.

A new Zoropsis species from China, with notes on Zoropsis pekingensis Schenkel, 1953 (Araneae, Zoropsidae).

 The family Zoropsidae is currently represented by 15 genera and 87 species (World Spider Catalog 2015). Zoropsids are medium to large size spiders, with a narrow, bipartite cribellum and calamistrum; eight eyes in two rows, anterior eye row nearly straight, posterior row strongly recurved; median furrow longitudinal; toothed chelicerae; tibiae and metatarsi I, II equipped with several pairs of ventral spines; metatarsi and tarsi bearing scopulae; two tarsal claws (Bosselaers 2002). They are often collected from houses, under stones or in leaf litter in forest.