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Purpose: To study the inhibitory effect of different doses of tetramethylpyrazine (TMP) combined with the beta-blocker, propranolol (Pro) on hemangioma endothelial (EOMA) cells. Methods: EOMA cells were cultured in vitro with varying doses of TMP and Pro (5, 10, 20 and 40 uM). The effect of treatments on cell proliferation was assessed by MTT assay, while cell apoptosis was assayed by flow cytometry. The expressions of Bcl-2, Bax, p-mTOR), total-mammalian target of rapamycin (t-mTOR, p-p70S6) and total-p70 ribosomal protein S6 (t-p70S6) proteins were determined using Western blot. Results: MTT data showed that when used alone, TMP had no significant inhibitory effect on EOMA cells (p > 0.05). However, when TMP was combined with propranolol, there was significant inhibition of EOMA cells, and that the inhibition is dependent on TMP dose. Flow cytometry results showed that the combination of TMP and Pro induced EOMA cell apoptosis dose-dependently (p < 0.05). Moreover, TMP dose-dependently inhibited the phosphorylation of mTOR and p70S6 in EOMA cells, and enhanced Bax expression, but downregulated Bcl-2 (p < 0.05). Conclusion: These results suggest that TMP enhances the inhibitory influence of Pro p-mTOR and pp-70S6 in EOMA cells in a dose-dependent manner. Thus, TMP may enhance Pro-induced inhibition of the growth of endothelial cells, and promote apoptosis through suppression of activation of PI3K/AKT signal route. These findings provide a theoretical basis for the clinical application of TMP/Pro combination for the treatment of hemangioma.

Hyunseong Kim,1 Jin Young Hong,1 Wan-Jin Jeon,1 Junseon Lee,1 Seung Ho Baek,2 In-Hyuk Ha1 1Jaseng Spine and Joint Research Institute, Jaseng Medical Foundation, Seoul, 135-896, Republic of Korea; 2College of Korean Medicine, Dongguk University, Goyang-si, 10326, Republic of KoreaCorrespondence: In-Hyuk HaJaseng Spine and Joint Research Institute, Jaseng Medical Foundation, Seoul, 135-896, Republic of KoreaTel +82-2-2222-2740Fax +82-2-527-1869Email hanihata@gmail.comPurpose: Lycopus lucidus Turcz (LLT) is a potent traditional medicinal herb that exerts therapeutic effects, regulating inflammatory disorders. However, the precise mechanisms by which LLT plays a potent role as an anti-inflammatory agent are still unknown, and in particular, the effects of LLT on cortical neurons and related mechanisms of neuroinflammation have not been studied. The NLRP3 inflammasome pathway is one of the most well known as an important driver of inflammation. We therefore hypothesized that LLT, as an effective anti-inflammatory agent, might have neurotherapeutic potential by inhibiting the NLRP3 inflammasome pathway in cortical neurons.Materials and Methods: Primary cortical neurons were isolated from the embryonic rat cerebral cortex, and H2O2 was used to stimulate neuron damage in vitro. After treatment with LLT at three concentrations (10, 25, and 50 μg/mL), the expression of iNOS, NLRP3, ASC, caspase-1, IL-1β, IL-18, IL-6, and IL-10 was determined by immunocytochemistry, qPCR, and ELISA. Neuron apoptosis was also evaluated using Annexin V-FITC/PI double staining FACS analysis. Neural regeneration-related factors (BDNF, NGF, synaptophysin, NT3, AKT, and mTOR) were analyzed by immunocytochemistry and qPCR.Results: LLT effectively protected cultured rat cortical neurons from H2O2-induced neuronal injury by significantly inhibiting NLRP3 inflammasome activation. In addition, it significantly reduced caspase-1 activation, which is induced by inflammasome formation and regulated the secretion of IL-1β/IL-18. We demonstrated that LLT enhances axonal elongation and synaptic connectivity upon H2O2-induced neuronal injury in rat primary cortical neurons.Conclusion: It was first demonstrated in vitro that LLT suppresses NLRP3 inflammasome activation, attenuates inflammation and apoptosis, and consequently promotes neuroprotection and the stimulation of neuron repair, suggesting that it is a promising therapeutic for neurological diseases.Keywords: Lycopus lucidus Turcz, cortical neuron, hydrogen peroxide, NLRP3 inflammasome, neuroinflammation

The current study tested the hypothesis of whether specific metal doping may show synergy with plant chromophore-based solar cells using a titanium dioxide (TiO2) electrode. A natural dye-sensitized, magnesium-doped TiO2 solar cell was assembled using the methanol extract from various western African plants including Lawsonia inermis (henna). Mg2+–TiO2 nanoparticles were applied on fluorine-doped tin oxide (FTO) glass to serve as the photoanode of the solar cells with $$I^{ - } /I_{3}^{ - }$$ electrolyte. A surface-modified TiO2 photoanode was prepared through the immersion method using a selective dopant including magnesium and potash. An inductively coupled plasma-optical emission spectrophotometer (ICP-OES) was utilized to characterize the potash dopant for comparative analysis. Instrumental analysis including ultraviolet–visible spectroscopy (UV–Vis), infrared spectroscopy (IR), and gas chromatography and mass spectrometry (GC–MS) analysis were carried out to characterize the natural henna dye extracts. The photovoltaic performance including open-circuit voltage (Voc), short circuit current density (Jsc), current (I), and power output (P) was analyzed quantitatively. ICP-OES analysis demonstrated that potash contains a composite of 26 elemental metals with K and Na accounting for 72.2% (5192.2 mg/kg) and 9.5% (682.6 mg/kg), respectively. GC–MS analysis confirmed the presence of lawsone and tocopherol in henna extracts. Among the tested samples, the Mg-doped TiO2 group generated the highest improvement in Jsc, from 0.66 to 1.28 (mA/cm2), representing a 93% increase. Our experiments demonstrated that the presence of magnesium as a doping agent improves the photogenerated electron transport and the light-harvesting performance of the henna dye to increase the overall efficiency of light-to-electricity conversion of the photovoltaic cells.

The current study tested the hypothesis of whether the low-temperature catalytic cracking of waste plastics would generate carbon fules using high-density polyethylene (HDPE) polymer by potash as a novel catalyst for both energy recovery and carbon recycling. We applied a one-stage pyrolysis reactor system with a 75 min reaction time to observe the highest yield at a low temperature range of 70–170 °C. The effects of the potash and zeolite catalysts, temperature, and catalyst–polymer ratio on the pyrolysis liquid yield and hydrocarbon contents were determined. The mineral concentration of potash was analyzed semiquantitatively using an inductively coupled plasma-optical emission spectrophotometer (ICP-OES) and X-ray diffractometer (XRD). The ICP-OES demonstrated that nine metals, in the order of K > Na > Fe > Si > Mg > Al > Cu > Ca > Ni, were predominant in the potash. GC–MS analysis of the liquid products showed that major catalytic cracking molecules are C11 to C20 as kerosene- to diesel-range liquid. Potash catalyst produced an average liquid conversion of 34.7% at a catalyst ratio of 30 wt% over a distillate temperature range of 76–140 °C, whereas zeolite generated 19.5% at the same catalyst ratio over 90–120 °C. Although the two catalysts favored mainly olefinic products, a higher potash ratio promoted a smaller carbon products with a purer composition. Our experiments demonstrated that the new natural potash catalyst could convert waste plastics into kerosene to diesel range of valuable and recyclable liquid products as potential renewable fuel sources for carbon recycling.

O-GlcNAc Transferase (OGT) is a complementary enzyme that regulates O-linked N-acetylglucosaminylation(O-GlcNAcylation) and plays a critical role in various cancer phenotypes, including invasion, migration, and metabolic reprogramming. In our previous study we found that miR-7-5p was downregulated at lung cancer cells with highly metastatic capacity. In the in-silico approach, OGT is the predicted target of miR-7-5p. To identify miR-7-5p′s role in cell growth and metabolism, we transfected various lung cancer cell lines with miR-7-5p. The expression level of miR-7-5p was confirmed by qRT-PCR in lung cancer cell lines. Western blot assays and qRT-PCR were performed to demonstrate miR-7-5p′s effect. Bioinformatic analysis indicated that OGT is a direct target of miR-7-5p. The binding sites of miR-7-5p in the OGT 3′ UTR were verified by luciferase reporter assay. To investigate the role of miR-7-5p in the cancer metabolism of non-small cell lung cancer (NSCLC) cells, mimic of miR-7-5p was transfected into NSCLC cells, and the effect of miR-7-5p on cancer metabolism was analyzed by LDH assays, glucose uptake, and mitochondrial ATP synthase inhibitor assay. O-GlcNAcylated protein level was determined by Western blot. The role of miR-7-5p in lung cancer growth was measured by MTS assays. To identify the delivery of miR-7-5p via PLGA, an in vitro release assay of PLGA-miR-7-5p was done. miR-7-5p was highly expressed whereas OGT showed low expression in H358, H827. However, miR-7-5p exhibited low expression while OGT had high expression in H522, H460, and H1299 cell lines. OGT were repressed by binding of miR-7a-5p to the 3′-UTR. Overexpression of miR-7-5p also diminished anaerobic glycolysis. miR-181a-5p transfection induced expression levels of OGT were diminished compared to those in the control group. O-GlcNAcylation was suppressed by miR-7-5p. Moreover, the overexpression of miR-7-5a suppressed lung cancer cell growth. miR-7-5p was released via PLGA for up to 10 days. In the present study, inhibition of OGT by miR-7-5p decreased the growth and cancer metabolism of lung cancer.

The current study examined the carbon recycling application of waste materials. Thermal catalytic cracking reactions were carried out in a fixed bed to synthesize gasoline-range hydrocarbon fuels from used plastics. Titanium (IV) oxide (TiO2) and zeolite were tested as catalysts for pyrolysis using low-density polyethylene (LDPE), polyvinylchloride (PVC), and polystyrene (PS) reactants. In addition to the catalyzed pyrolysis reactions, we also investigated non-catalyzed thermal degradation of the plastic substrates for negative control. The liquid yield, reaction temperature profile, and physical appearance of the synthesized liquid products were determined. The pyrolysis reactions demonstrated that the optimum catalyst–polymer ratio is 40%. The distillate collection temperatures ranged between 82 and 198 °C (LDPE), 68–172 °C (PVC), and 40–168 °C (PS). Our experiments showed that LDPE, PVC, and PS can readily be pyrolyzed to produce 44% (LDPE), 13% (PVC), and 89% (PS) hydrocarbon liquid products using zeolite catalyst. Gas chromatography–mass spectrometry (GC–MS) was used to analyze the structure and chemical composition of the products. The main products were C5 (1,2-dimethylcyclopropane), C6 (2-methylpentane), C7 (1,3-dimethylcyclopentene, 1-heptene), and C8 (2-octene, 4-octene, octane, 3-ethylhexane), indicating gasoline-range hydrocarbon molecules. The highest liquid yield of 89.3% was obtained from zeolite catalyst over polystyrene in comparison to all plastics cracked while the lowest liquid yield of 3.9% was obtained from the cracking of PVC under no catalyst condition.

Constant or intense light degenerates the retina and retinal pigment epithelial cells. Light generates reactive oxygen species and nitric oxide leading to initial reactions of retinal degeneration. Apoptosis is the primary mechanism of abnormal death of photoreceptors, retinal ganglion cells, or retinal pigment epithelium (RPE) in degenerative retinal diseases, including diabetic retinopathy and age-related macular degeneration. The current study evaluated the function of erythropoietin (EPO) on angiogenesis and apoptosis in the retina and RPE under oxidative stress. We determined the pro-angiogenic and antiapoptotic mechanism of EPO under stress conditions using a conditional EPO knockdown model using siRNA, EPO addition, proteomics, immunocytochemistry, and bioinformatic analysis. Our studies verified that EPO protected retinal cells from light-, hypoxia-, hyperoxia-, and hydrogen peroxide-induced apoptosis through caspase inhibition, whereas up-regulated angiogenic reactions through vascular endothelial growth factor (VEGF) and angiotensin pathway. We demonstrated that the EPO expression in the retina and subsequent serine/threonine/tyrosine kinase phosphorylations might be linked to oxidative stress response tightly to determining angiogenesis and apoptosis. Neuroprotective roles of EPO may involve the balance between antiapoptotic and pro-angiogenic signaling molecules, including BCL-xL, c-FOS, caspase-3, nitric oxide, angiotensin, and VEGF receptor. Our data indicate a new therapeutic application of EPO toward retinal degeneration based on the dual roles in apoptosis and angiogenesis at the molecular level under oxidative stress.

PURPOSE: The current study aimed to determine the antidiabetic effects of leaf extract of Ficus asperifolia in streptozotocin-induced diabetic rats. METHODS: A total of ninety-five (95) adult rats were used for the experiment. The whole study protocol was divided into three sets of individual experiments. The animals were divided randomly into seven groups of five rats each. The rats were given a diet supplemented with 50 g high fat to 50 g vital feeds for two weeks. The study lasted 28 days with daily administration and weekly blood glucose and body weight check. At the end of the experiment protocol, the rats were fasted overnight and were anesthetized. Blood was collected via cardiac puncture from each animal for biochemical analysis. Metglim 3.38 mg/kg bodyweight was used as positive control. Diabetes was induced using streptozotocin (35 mg/kg in 0.1 M phosphate-buffered saline) intraperitoneally for 5 days. Phytochemicals were analyzed in both extract and fractions. RESULTS: Phytochemical screening revealed the presence of alkaloids, saponins, flavonoids, glycosides, tannins, carotenoids, terpenes, and steroids in both extract and fractions. Proteins, carbohydrates, and fats were detected by systematic molecular analysis. Fraction 1 of plant extracts prevented glucose-induced hyperglycaemia 30 min’ post glucose load in all rats. Streptozotocin treatment caused a significant increase (p˂0.05) in blood sugar, total cholesterol, triacylglycerol, low-density lipoproteins and a significant (p˂0.05) decrease in food intake, body weight and high-density lipoproteins in diabetic rats. CONCLUSION: Treatment with the extract significantly improved the derangements caused by streptozotocin. These results suggest that the leaf extracts of Ficus asperifolia could serve as a potential treatment for diabetes therapy. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s40200-020-00524-1) contains supplementary material, which is available to authorized users.

Gongjin-dan (GJD) is a multiherbal formula produced from 10 medicinal herbs and has been traditonally used as an oriental medicine to treat cardiovascular diseases, alcoholic hepatitis, mild dementia, and anemia. Additionally, increasing evidence suggests that GJD exerts neuroprotective effects by suppressing inflammation and oxidative stress-induced events to prevent neurological diseases. However, the mechanism by which GJD prevents oxidative stress-induced neuronal injury in a mature neuron remains unknown. Here, we examined the preventive effect and mechanism of GJD on primary cortical neurons exposed to hydrogen peroxide (H2O2). In the neuroprotection signaling pathway, Sirtuin1 is involved in neuroprotective action as a therapeutic target for neurological diseases. After pre-treatment with GJD at three concentrations (10, 25, and 50 µg/mL) and stimulation by H2O2 (30 µM) for 24 h, the influence of GJD on Sirtuin1 activation was assessed using immunocytochemistry, real-time PCR, western blotting, and flow cytometry. GJD effectively ameliorated H2O2-induced neuronal death against oxidative damage through Sirtuin1 activation. In addition, GJD-induced Sirtuin1 activation accelerated elongation of new axons and formation of synapses via increased expression of nerve growth factor and brain-derived neurotrophic factor, as well as regeneration-related genes. Thus, GJD shows potential for preventing neurological diseases via Sirtuin1 activation.

Objective To optimize a self-emulsifying drug delivery system (SEDDS) formulation for psoralen and isopsoralen (PSO and IPSO) isolated from Psoraleae Fructus. Methods A D-optimal design was used to investigate the influence of oil percentage, surfactant percentage and cosurfactant percentage on several properties of SEDDS including particle size, polydispersity, equilibrium solubility, in situ intestine absorption rate and intestinal permeability. Furthermore, the desirability function approach was applied to obtain the optimal formulation for the system. Results The oil percentage, surfactant percentage and cosurfactant percentage were optimized to be 53.6%, 35.7% and 10.7%, respectively, which means the model is available. Conclusions The D-optimal design is valuable to optimize the SEDDS formulation and understand formulation compositions’ functions on SEDDS properties.

1D hollow Co0.5Mn0.5Fe2O4-PANI nanofibers were synthesized through sequential processes of electrospinning, heat treatment in air, and chemical polymerization of polyaniline. The morphology and photocatalytic properties of heterostructured Co0.5Mn0.5Fe2O4-PANI as photocatalysts were investigated by SEM, TEM, XRD, TGA, and photocatalysis experiments of methyl orange under visible-light irradiation. The hollow Co0.5Mn0.5Fe2O4-PANI nanofibers showed a high photocatalytic degradation efficiency of 92% within 120 min and a kinetic constant of 115-times higher than hollow Co0.5Mn0.5Fe2O4 nanofibers. These high photocatalytic properties are attributed to both unique 1D hollow nanofiber morphology with macroporous hollow core, PANI/Co0.5Mn0.5Fe2O4/PANI shell and the heterojunction structure composed of Co0.5Mn0.5Fe2O4 and PANI. In addition, the outstanding magnetic properties of the hollow Co0.5Mn0.5Fe2O4-PANI nanofibers with its inherent spinel structure facilitate the recovery of the photocatalyst.

Benzothieno[60]fullerenes were synthesized using fullerenyl cations as key intermediates. The reaction proceeded through a nucleophilic attack of the sulfur atom as a weak nucleophile to the fullerenyl cation electrophile. A monoarylated fullerene, (2-methylthiophenyl)hydro[60]fullerene, C

Apamin is a minor component of bee venom and is a polypeptide with 18 amino acid residues. Although apamin is considered a neurotoxic compound that blocks the potassium channel, its neuroprotective effects on neurons have been recently reported. However, there is little information about the underlying mechanism and very little is known regarding the toxicological characterization of other compounds in bee venom. Here, cultured mature cortical neurons were treated with bee venom components, including apamin, phospholipase A2, and the main component, melittin. Melittin and phospholipase A2 from bee venom caused a neurotoxic effect in dose-dependent manner, but apamin did not induce neurotoxicity in mature cortical neurons in doses of up to 10 µg/mL. Next, 1 and 10 µg/mL of apamin were applied to cultivate mature cortical neurons. Apamin accelerated neurite outgrowth and axon regeneration after laceration injury. Furthermore, apamin induced the upregulation of brain-derived neurotrophic factor and neurotrophin nerve growth factor, as well as regeneration-associated gene expression in mature cortical neurons. Due to its neurotherapeutic effects, apamin may be a promising candidate for the treatment of a wide range of neurological diseases.

Lumbar spinal stenosis (LSS) is a common neurodegenerative condition. However, how neurogenic claudication develops with severe leg pain has not yet been clearly elucidated. Moreover, cerebrospinal fluid (CSF) physiology at the lumbosacral level is poorly understood because of the difficulties involved in quantification and visualization. Recent studies have suggested that assessment of mitochondrial function in CSF provides an indirect way to assess neurological disorders and an important feature of disease progression. In this study, we assessed the relevance of endogenous extracellular mitochondria in the CSF of rats after LSS. Mitochondrial changes within the CSF were analyzed following LSS at 1 week using flow cytometry. An increase in cell size and number was observed in CSF with LSS, and reactive oxygen species (ROS) levels were also increased within the CSF at 1 week in the LSS group. Elevated mitochondrial ROS and functional changes in the CSF are hallmarks of LSS. The present study is the first to demonstrate that elevated mitochondrial ROS within the CSF is a new index for the early detection of LSS. Moreover, it may represent a potential novel treatment target for LSS.

Inula britannica var. chinensis (IBC) has been used as a traditional medicinal herb to treat inflammatory diseases. Although its anti-inflammatory and anti-oxidative effects have been reported, whether IBC exerts neuroprotective effects and the related mechanisms in cortical neurons remain unknown. In this study, we investigated the effects of different concentrations of IBC extract (5, 10, and 20 µg/mL) on cortical neurons using a hydrogen peroxide (H2O2)-induced injury model. Our results demonstrate that IBC can effectively enhance neuronal viability under in vitro-modeled reaction oxygen species (ROS)-generating conditions by inhibiting mitochondrial ROS production and increasing adenosine triphosphate level in H2O2-treated neurons. Additionally, we confirmed that neuronal death was attenuated by improving the mitochondrial membrane potential status and regulating the expression of cytochrome c, a protein related to cell death. Furthermore, IBC increased the expression of brain-derived neurotrophic factor and nerve growth factor. Furthermore, IBC inhibited the loss and induced the production of synaptophysin, a major synaptic vesicle protein. This study is the first to demonstrate that IBC exerts its neuroprotective effect by reducing mitochondria-associated oxidative stress and improving mitochondrial dysfunction.

There are many epidemiological studies asserting that fine dust causes lung cancer, but the biological mechanism is not clear. This study was conducted to investigate the effect of PM10 (particulate matter less than 10 &mu, m) on single nucleotide variants through whole genome sequencing in lung epithelial cancer cell lines (HCC-827, NCI-H358) that have been exposed to PM10. The two cell lines were exposed to PM10 for 15 days. We performed experimental and next generation sequencing analyses on experimental group that had been exposed to PM10 as well as an unexposed control group. After exposure to PM10, 3005 single nucleotide variants were newly identified in the NCI-H358 group, and 4402 mutations were identified in the HCC-827 group. We analyzed these single nucleotide variants with the Mutalisk program. We observed kataegis in chromosome 1 in NCI-H358 and chromosome 7 in HCC-827. In mutational signatures analysis, the COSMIC mutational signature 5 was highest in both HCC-827 and NCI-H358 groups, and each cosine similarity was 0.964 in HCC-827 and 0.979 in the NCI-H358 group. The etiology of COSMIC mutational signature 5 is unknown at present. Well-designed studies are needed to determine whether environmental factors, such as PM10, cause COSMIC mutational signature 5.

Activation of the Hedgehog pathway may have therapeutic value for improved bone healing, taste receptor cell regeneration, and alleviation of colitis or other conditions. Systemic pathway activation, however, may be detrimental, and agents amenable to tissue targeting for therapeutic application have been lacking. We have developed an agonist, a conformation-specific nanobody against the Hedgehog receptor Patched1 (PTCH1). This nanobody potently activates the Hedgehog pathway in vitro and in vivo by stabilizing an alternative conformation of a Patched1 "switch helix," as revealed by our cryogenic electron microscopy structure. Nanobody-binding likely traps Patched in one stage of its transport cycle, thus preventing substrate movement through the Patched1 sterol conduit. Unlike the native Hedgehog ligand, this nanobody does not require lipid modifications for its activity, facilitating mechanistic studies of Hedgehog pathway activation and the engineering of pathway activating agents for therapeutic use. Our conformation-selective nanobody approach may be generally applicable to the study of other PTCH1 homologs.

Inflammation is an essential biological response that eliminates pathogenic bacteria and repairs tissue after injury. Acute kidney injury (AKI) is associated with systemic and intrarenal inflammation as the inflammatory process decreases renal function and promotes progression to advanced chronic kidney disease. Macrophages are key mediators of the inflammatory response, their activation influences the immune system and may have various effects. Classically activated type I macrophages (M1) produce a variety of pro-inflammatory cytokines at the lesion site. However, anti-inflammatory type II macrophages (M2) are alternatively activated upon exposure to anti-inflammatory cytokines and are associated with wound healing and tissue repair following AKI. Here, we used melittin from bee venom to enhance the polarization of M2 macrophages and promote renal recovery after AKI. Melittin was administered to mice intraperitoneally for 5 days at various concentrations (10, 50, and 100 &micro, g/kg), serum creatinine and blood urea nitrogen (BUN) levels were analyzed 72 h after cisplatin administration to confirm renal dysfunction. Melittin inhibited the cisplatin-induced increase in creatinine and BUN, an indicator of renal dysfunction. The expression of M1 markers (CD16/32) decreased significantly, whereas that of M2 markers (CD206, Arg1nase I) increased after melittin administration. Consistently, tubular necrosis was substantially reduced in melittin-treated mice. Thus, melittin alleviates cisplatin-induced AKI by regulating M2 macrophage expression.

Oxidative stress plays a key role in the pathogenesis of several diseases, including neurodegenerative diseases. Recent studies have reported that mitochondrial dysfunction is a leading cause of the overproduction of reactive oxygen species and oxidative stress. Mitochondrial changes play an important role in preventing oxidative stress. However, there is a lack of experimental evidence supporting this hypothesis. Thymus quinquecostatus CELAK (TQC) extract is a plant from China belonging to the thymus species, which can mediate the inflammatory response and prevent cell damage through its antioxidant activities. This study examines whether TQC can scavenge excess ROS originating from the mitochondria in RAW 264.7 macrophages. We used lipopolysaccharide (LPS) to induce inflammation and oxidative stress in RAW 264.7 macrophages and performed an immunocytochemistry dot blot of 8-hydroxy-2&prime, deoxyguanosine (8-OHdG) and real-time PCR to analyze the expression levels of genes involved in mitochondrial biogenesis and oxidative metabolism. TQC was found to significantly reduce the intensity of immunostained MitoSOX and 8-OHdG levels in the total genomic DNA within the mitochondria in RAW 264.7 macrophages. The HO-1 and Nrf2 mRNA levels were also significantly increased in the TQC groups. Therefore, we verified that TQC improves mitochondrial function and attenuates oxidative stress induced by LPS. Our results can provide reference for the effect of TQC to develop new therapeutic strategies for various diseases.

Aims CREB (cAMP response element-binding protein)-regulated transcription coactivator (CRTC2) has been reported to act as a coactivator of CREB during gluconeogenesis. The role of CRTC2 in osteoblastic differentiation has not yet been elucidated. The aim of this study is to identify the mechanism of CRTC2 in osteoblast differentiation. Main methods The mRNA expression was determined by RT-PCR and qPCR. Protein levels were measured using Western blot assay. Alkaline phosphatase (ALP) staining was performed to evaluate ALP activity. Alizarin red S (ARS) staining was performed to measure extracellular mineralization. Transcriptional activity was detected using a luciferase assay. Key findings In the present study, TNF-α was found to stimulate CRTC2 expression. However, TNF-α did not increase the gene expression of osteoblast differentiation markers and inhibited BMP2-induced osteoblastic differentiation. Overexpression of CRTC2 decreased the expression of osteogenic genes, ALP activity and extracellular matrix mineralization. Knockdown of CRTC2 restored BMP2-induced osteogenic gene expression and ALP activity. CRTC2 increased Smurf1 mRNA expression, Smurf 1 promoter activity, and protein level. Furthermore, Smurf 1 decreased Smad 1/5/9 protein levels. These results suggest that CRTC2 decreased BMP2-induced osteoblastic differentiation via Smurf 1 expression. Significance Our results indicate that CRTC2 regulates the expression of Smurf1 in osteoblast differentiation.

Parkinson’s disease (PD) is a neurodegenerative disease seriously threatening the elderly. Recent studies have shown that autophagy plays a key role in the clearance of organelles associated with neurodegenerative diseases, and plays an important role in the development and progression of PD; however, its mechanism is not yet clear. In this study, the model of aging mice was used to establish PD model by intra peritoneal injection of MPTP to study the effect of aging changes of autophagy on the mechanism of MPTP toxicity mechanism and its relationship with mitochondrial function and mitochondrial dynamics; and at the same time, young mice were selected to carry out moderate-intensity treadmill training, after 6 weeks, the PD model was established to explore the effect of exercise pre-training on the above indexes of PD mice, and explore the relationship between autophagy and PD, autophagy and mitochondrial dynamics and mitochondrial function.

Thermo-reactive deposition (TRD) salt bath Nb-Cr co-diffusion process was brought forward to prepare the functionally gradient chromium-niobium carbide coating. In TRD salt bath Nb-Cr co-diffusion treatment, T10 steel samples were delved into TRD salt. The samples were treated at 880°C for 3.5 h and then at 950°C for 2.5h. The obtained coatings were characterized by means of microstructure, microhardness, scanning electron microscopy (SEM), energy dispersive X-ray spectrometry (EDX) and X-ray diffraction analysis (XRD). The cross-section hardness of the chromium-niobium carbide coating obtained on the surface of T10 steel samples transits smoothly. The coating was dense, smooth and compact. The thickness ratio of the niobium carbide layer (the outer layer) and the chromium carbide layer (the inner layer) was 4. The hardness of chromium-niobium carbide coating was about 2751HV0.05, which is between the single chromizing and single niobium. The results showed superior wear properties of the coated samples. The kinetics of chromium-niobium carbide coating was also studied.

The retinal pigment epithelium (RPE) plays imperative roles in normal retinal function by photoreceptor protection from light and phagocytosis of rod and cone outer segments during disc shedding. Melatonin is the free radical scavenger and circadian determinant to protect the RPE and retina from oxidative stress and regulate the circadian clock. The current study tested the hypothesis whether melatonin could affect cytoskeletal structure within RPE. Our Western blot analysis demonstrated that melatonin treatment up-regulated prohibitin 3-fold compared to control. β-tubulin levels were also up-regulated by melatonin but to a lesser extent. Initial cell shape of ARPE-19 is epitheloid, however, after 30-minute treatment with melatonin, RPE cells undergo a morphological change to a fusiform shape with spindle outgrowth. Cells return to epitheloid shape after 12 hours in untreated medium. Melatonin treated cells showed increased and dissimilar distribution of prohibitin and β-tubulin compared to non-treated cells, thus altered cytoskeletal and mitochondrial structure in the RPE. Our data implies that melatonin may play a protective role under oxidative stress, which is shown by the marker prohibitin in terms of increased expression and nuclear distribution. During the protective process, cells change their morphology. Our results suggest that melatonin treatment could be beneficial to protect mitochondria under oxidative stress and treat certain ocular diseases, including age-related macular degeneration.

As organic photodetectors with less than 1 μm pixel size are in demand, a new way of enhancing the sensitivity of the photodetectors is required to compensate for its degradation due to the reduction in pixel size. Here, we used Ag nanoparticles coated with SiOxNy as a light-absorbing layer to realize the scale-down of the pixel size without the loss of sensitivity. The surface plasmon resonance appeared at the interface between Ag nanoparticles and SiOxNy. The plasmon resonance endowed the organic photodetector with boosted photon absorption and external quantum efficiency. As the Ag nanoparticles with SiOxNy are easily deposited on ITO/SiO2, it can be adapted into various organic color image sensors. The plasmon-supported organic photodetector is a promising solution for realizing color image sensors with high resolution below 1 μm.

Activation of the Hedgehog pathway may have therapeutic value for improved bone healing, taste receptor cell regeneration, and alleviation of colitis or other conditions. Systemic pathway activation, however, may be detrimental and therapeutic application has been difficult for lack of agents amenable to tissue targeting. We have developed a novel agonist, a conformation-specific nanobody against the Hedgehog receptor Patched1. This nanobody potently activates the Hedgehog pathway in vitro and in vivo by stabilizing an alternative conformation of a Patched1 “switch helix”, as revealed by cryo-EM structure determination. Although this conformation likely constitutes part of the transport cycle, nanobody-trapping disrupts the cycle and prevents substrate movement through the Patched1 sterol conduit. Our conformation-selective nanobody approach provides a new route to the development of transporter-related pharmacologic agents and may be generally applicable to the study of other transporters.

A method to directly collect negatively charged nucleic acids, such as DNA and RNA, in the biosamples simply by applying an electric field in between the sample and collection buffer separated by the nanofilter membrane is proposed. The nanofilter membrane was made of low-stress silicon nitride with a thickness of 100 nm, and multiple pores were perforated in a highly arranged pattern using nanoimprint technology with a pore size of 200 nm and a pore density of 7.22 × 108/cm2. The electrophoretic transport of hsa-mir-93-5p across the membrane was confirmed in pure microRNA (miRNA) mimic solution using quantitative reverse transcription-polymerase chain reactions (qRT-PCR). Consistency of the collected miRNA quantity, stability of the system during the experiment, and yield and purity of the prepared sample were discussed in detail to validate the effectiveness of the electrical protocol. Finally, in order to check the applicability of this method to clinical samples, liquid biopsy process was demonstrated by evaluating the miRNA levels in sera of hepatocellular carcinoma patients and healthy controls. This efficient system proposed a simple, physical idea in preparation of nucleic acid from biosamples, and demonstrated its compatibility to biological downstream applications such as qRT-PCR as the conventional nucleic acid extraction protocols.

The current study tested the hypothesis whether common plant extracts could be used as redox molecules in biobattery. Natural quinone molecules were extracted from Lawsonia inermis (henna) via sequential extraction using hexane, ethyl acetate, methanol and 80% methanol in water, followed by purification using column chromatography to examine their potential function as redox molecules in biobattery. A combination of UV–visible spectroscopy and gas chromatography–mass spectrometry (GC–MS) analysis confirmed the presence of quinones in the extracted fractions. UV analysis showed maximum absorbance at 295 nm and 450 nm which correspond to 4-t-butyl-1,2-benzoquinone and duroquinone. In addition, GC–MS analysis of the henna extract confirmed the presence of tocopherol (vitamin E) as a potential redox molecule. We determined the impact of the type of electrolyte, electrode, salt bridge and volume of extract on the overall efficiency of biobattery. Among the different cell combinations tested, the optimum battery with a maximum voltage of 0.97 V was achieved using a carbon||quinone cathodic half-cell, copper||sulphuric acid anodic half-cell and a KCl (1.0 M) salt bridge. Our experiments demonstrate that natural redox molecules from common African plants, including L. inermis extracts, can serve as source of electrical energy and alternative materials for the renewable battery.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease with a wide range of survival times. We aimed to explore prognostic factors related to short survival based on clinical features and plasma metabolic signatures using surface-enhanced Raman spectroscopy (SERS). One hundred and thirty-eight sporadic ALS cases were enrolled serially, including 62 for the short-duration group (≤3 years) and 76 for the long-duration group (3 years). Multivariate analysis showed that an older age of onset (60 years; odds ratio [OR] = 3.98, 95% CI: 1.09-14.53), lower body mass index (BMI) (18.5; OR = 6.80, 95% CI: 1.36-33.92), and lower ALSFRS-R score (35; OR = 6.03, 95% CI: 1.42-25.63) were associated with higher odds of tracheotomy or death, while a higher uric acid (UA) level showed a protective effect (356.36 μmol/L; OR = 0.19, 95% CI: 0.05-0.73). SERS analysis showed significant differences between the two groups, and pathway analysis highlighted five main metabolic pathways, including metabolisms of glutathione, pyrimidine, phenylalanine, galactose, and phenylalanine-tyrosine-tryptophan biosynthesis. In conclusion, age of onset, BMI, ALSFRS-R score and UA, together with dysregulation of glucose, amino acid, nucleic acid, and antioxidant metabolism contributed to disease progression, and are therefore potential therapeutic targets for ALS.

Lumbar spinal stenosis (LSS), one of the most commonly reported spinal disorders, can cause loss of sensation and dyskinesia. In currently used animal models of LSS, the spinal cord is covered entirely with a silicone sheet, or block-shaped silicone is inserted directly into the spinal canal after laminectomy. However, the effects of differences between these implant materials have not been studied. We assessed the degree of damage and locomotor function of an LSS model in Sprague-Dawley rats using silicone blocks of varying hardness (70, 80, and 90 kPa) implanted at the L4 level. In sham rats, the spinal cord remained intact; in LSS rats, the spinal cord was increasingly compressed by the mechanical pressure of the silicone blocks as hardness increased. Inflammatory cells were not evident in sham rats, but numerous inflammatory cells were observed around the implanted silicone block in LSS rats. CD68+ cell quantification revealed increases in the inflammatory response in a hardness-dependent manner in LSS rats. Compared with those in sham rats, proinflammatory cytokine levels were significantly elevated in a hardness-dependent manner, and locomotor function was significantly decreased, in LSS rats. Overall, this study showed that hardness could be used as an index to control the severity of nerve injury induced by silicone implants.

Abnormal vessel formations contribute to the progression of specific angiogenic diseases including age-related macular degeneration. Adequate vessel growth and maintenance represent the coordinated process of endothelial cell proliferation, matrix remodeling, and differentiation. However, the molecular mechanism of the proper balance between angiogenic activators and inhibitors remains elusive. In addition, quantitative analysis of vessel formation has been challenging due to complex angiogenic morphology. We hypothesized that conjugated double bond containing-natural products, including anthocyanin extracts from Hibiscus sabdariffa, may control the proper angiogenesis. The current study was designed to determine whether natural molecules from African plant library modulate angiogenesis. Further, we questioned how the proper balance of anti- or pro-angiogenic signaling can be obtained in the vascular microenvironment by treating anthocyanin or fatty acids using chick chorioallantoic membrane angiogenesis model in ovo. The angiogenic morphology was analyzed systematically by measuring twenty one angiogenic indexes using Angiogenic Analyzer software. Chick chorioallantoic model demonstrated that anthocyanin-rich extracts inhibited angiogenesis in time- and concentration-dependent manner. Molecular modeling analysis proposed that hibiscetin as a component in Hibiscus may bind to the active site of vascular endothelial growth factor receptor 2 (VEGFR2) with ΔG= −8.42 kcal/mol of binding energy. Our results provided the evidence that anthocyanin is an angiogenic modulator that can be used to treat uncontrolled neovascular-related diseases, including age-related macular degeneration.

The current study aims to determine the molecular mechanisms of age-related macular degeneration (AMD) using the phosphorylation network. Specifically, we examined novel biomarkers for oxidative stress by protein interaction mapping using in vitro and in vivo models that mimic the complex and progressive characteristics of AMD. We hypothesized that the early apoptotic reactions could be initiated by protein phosphorylation in region-dependent (peripheral retina vs. macular) and tissue-dependent (retinal pigment epithelium vs. retina) manner under chronic oxidative stress. The analysis of protein interactome and oxidative biomarkers showed the presence of tissue- and region-specific post-translational mechanisms that contribute to AMD progression and suggested new therapeutic targets that include ubiquitin, erythropoietin, vitronectin, MMP2, crystalline, nitric oxide, and prohibitin. Phosphorylation of specific target proteins in RPE cells is a central regulatory mechanism as a survival tool under chronic oxidative imbalance. The current interactome map demonstrates a positive correlation between oxidative stress-mediated phosphorylation and AMD progression and provides a basis for understanding oxidative stress-induced cytoskeletal changes and the mechanism of aggregate formation induced by protein phosphorylation. This information could provide an effective therapeutic approach to treat age-related neurodegeneration.

A novel approach for the thin film encapsulation (TFE) of organic photo-diode (OPD) for the next generation of organic/inorganic hybrid complementary metal oxide semiconductor (CMOS) image sensor is reported. The TFE is composed of two different metal-oxides stacked in bi-layer thin film architecture. The first layer is composed of aluminum oxide (AlOx) deposited by atomic layer deposition (ALD) at a moderate temperature of 100 °C to avoid any damage to the organic active layer. The first layer acts as a water barrier layer and also as a first protective layer for the deposition of a second silicon oxynitride (SiON) layer that could be processed by plasma-enhanced chemical vapor deposition (PECVD) at higher temperatures. The second layer ensures a better mechanical and chemical stability of the whole structure and also serves as a second protective layer from damages induced during the additional processing stages, such as photolithography or microlensing. With the TFE architecture the overall device stability at 85 °C and 85% relative humidity exceeded 1000 h without observable device performance decrease. This was confirmed by fabricating a green-light sensitive OPD characterized by a stable external quantum efficiency of 60–70%.

In vertebrates, myeloid cells arise from multiple waves of development: the first or embryonic wave of myelopoiesis initiates early from non-hematopoietic stem cell (HSC) precursors and gives rise to myeloid cells transiently during early development; whereas the second or adult wave of myelopoiesis emerges later from HSCs and produces myeloid cells continually during fetal and adult life. In the past decades, a great deal has been learnt about the development of myeloid cells from adult myelopoiesis, yet the genetic network governing embryonic myelopoiesis remains poorly defined. In this report, we present an in vivo study to delineate the role of Cebpα during zebrafish embryonic myelopoiesis. We show that embryonic myelopoiesis in cebpα-deficient zebrafish mutants initiates properly but fails to produce macrophages and neutrophils. The lack of macrophages and neutrophils in the mutants is largely attributed to the cell cycle arrest of embryonic myeloid progenitors, resulting in the impairment of their maintenance and subsequent differentiation. We further show that Cebpα, perhaps acting cooperatively with Runx1, plays a critical role in embryonic neutrophil maintenance. Our findings reveal a new role of Cebpα in embryonic myelopoiesis.

The study of the optical properties of pristine BNNT, Si-doped BNNTs and SO2 molecule adsorption on Si-doped BNNTs is that, to our knowledge, few relevant research have ever been found. In this paper, the adsorption behaviors of Sulfur dioxide (SO2) molecule on Si-doped Boron nitride nanotubes (BNNTs) are investigated applying the first-principles calculations. The main contribution of this paper is that the foremost investigation for the optical properties of the pristine BNNT, Si-doped BNNTs and SO2 adsorption on Si-doped BNNTs. Additionally, the electronic properties and the structural properties are also presented. In our calculations of optical properties, the dielectric constant, the refractive index and the absorption coefficient are obtained. Comparing the pristine BNNT, our results indicate that, the blue-shifts (in the main peaks of the dielectric constant of Si B - BNNT and SO 2 - Si B - BNNT ), and the red-shifts (in the main peaks of the refractive index of Si N - BNNT and SO 2 - Si N - BNNT ) are appeared. Under these conditions, Si-doped BNNT and Si-doped BNNT with SO2 adsorption, the gaps are reduced both for the speculated optical band gaps and the electronic structure band gaps.

Microglia are CNS-resident macrophages and play important roles in neural development and function. However, how microglial precursors born in peripheral tissues colonize the CNS remains undefined. Using in vivo imaging and genetic manipulation of zebrafish, we showed that microglial precursors enter the optic tectum of the midbrain, where the majority of microglia reside during early development, via the lateral periphery between the eyes and brain and the ventral periphery of the brain in a circulation-independent manner. The colonization of the optic tectum by microglial precursors is dynamic and driven by apoptotic neuronal death, which occurs naturally in the midbrain during neurogenesis. We further show that lysophosphatidylcholine, a phospholipid known to be released from apoptotic cells, can promote microglial precursor entry into the brain via its cognate receptors grp132b. Our study reveals that microglia colonization of developing zebrafish midbrain is triggered by apoptotic neuronal death, possibly via releasing lysophosphatidylcholine.

Hollow cobalt ferrite–polyaniline nanofibers (CoFe 2 O 4 –PANI) functioning as a photocatalyst were prepared by using electrospinning technique, followed by calcination and in-situ chemical oxidative polymerization. The hollow CoFe 2 O 4 –PANI nanofibers with one dimensional (1D) structure provide well-developed mesoporous structure and enhanced electrical conductivity. The hollow CoFe 2 O 4 –PANI nanofibers consisted of hollow core-double shell nanostructure, which has CoFe 2 O 4 as inner shell and PANI as outer shell. The hollow CoFe 2 O 4 –PANI nanofibers coated with PANI nanograins facilitate the optical properties by easily capturing visible light. The intensive visible light absorption and effective charge separation lead to a remarkable improvement of visible light photocatalysis owing to the heterojunction built between CoFe 2 O 4 and PANI. The pseudo-first-order kinetic constant of photocatalytic degradation of the methyl orange (MO) dye under visible light irradiation with CoFe 2 O 4 –PANI was 80 times greater than CoFe 2 O 4 . This evidence shows the advantage of the unique core–shell mesoporous structure in the CoFe 2 O 4 –PANI for efficient photocatalysis. The recovery ratio is almost the same after three cycles under visible light irradiation, displaying the excellent characteristics of magnetic separation for CoFe 2 O 4 –PANI hollow nanofibers.

Cyclic GMP-AMP synthase (cGAS) is a key sensor responsible for cytosolic DNA detection. Here we report that GTPase-activating protein SH3 domain–binding protein 1 (G3BP1) is critical for DNA sensing and efficient activation of cGAS. G3BP1 enhanced DNA binding of cGAS by promoting the formation of large cGAS complexes. G3BP1 deficiency led to inefficient DNA binding by cGAS and inhibited cGAS-dependent interferon (IFN) production. The G3BP1 inhibitor epigallocatechin gallate (EGCG) disrupted existing G3BP1–cGAS complexes and inhibited DNA-triggered cGAS activation, thereby blocking DNA-induced IFN production both in vivo and in vitro. EGCG administration blunted self DNA–induced autoinflammatory responses in an Aicardi–Goutieres syndrome (AGS) mouse model and reduced IFN-stimulated gene expression in cells from a patient with AGS. Thus, our study reveals that G3BP1 physically interacts with and primes cGAS for efficient activation. Furthermore, EGCG-mediated inhibition of G3BP1 provides a potential treatment for cGAS-related autoimmune diseases. Li and colleagues show that G3BP1, a protein known to regulate the RNA stress response, is critical for DNA sensing and efficient activation of the cytoplasmic DNA sensor cGAS.

Controlling defect states in a buffer layer for organic photo devices is one of the vital factors which have great influence on the device performance. Defect states in silicon oxynitride (SiOxNy) buffer layer for organic photo devices can be controlled by introducing appropriate dopant materials. We performed ab initio simulations to identify the effect on doping SiOxNy with carbon (C), boron (B), and phosphorous (P) atoms. The results unveil that hole defects in the SiOxNy layer diminish with the phosphorous doping. Based on the simulation results, we fabricate the small molecule organic photodetector (OPD) including the phosphorous-doped SiOxNy buffer layer and the active film of blended naphthalene-based donor and C60 acceptor molecules, which shows excellent enhancement in the external quantum efficiency (EQE). The results of our charge-based deep level transient spectroscopy (Q-DLTS) measurements confirmed that the EQE enhancement originates from the decrease of the hole traps induced by the reduced hole defects. The method of controlling the defect states in SiOxNy buffer layers by the doping can be used to improve the performance in various organic photo devices.