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Abstract Background Paraplegia after spinal cord ischemia is a devastating condition in the clinic. Here, we develop an awake rabbit model of spinal cord ischemia with delayed paraplegia and explore the influence of ambient temperature on the outcomes after injury. Methods A total of 47 male rabbits were involved in the present study. Transient spinal cord ischemia was induced by occluding the infrarenal abdominal aorta of awake rabbits at different ambient temperatures. To find the optimal conditions for developing delayed paraplegia, hindlimb motor function after ischemia was evaluated between experiments. Results The onset and magnitude of ischemic injury varied with the ambient temperature maintained during the peri‐ischemia period. More serious spinal cord injury occurred when ischemia was induced at higher temperatures. At 18°C, 25‐minute ischemia resulted in 74% of rabbits developing delayed paraplegia. At a temperature of 28°C or higher, most of the animals developed acute paraplegia immediately. While at 13°C, rabbits usually regained normal motor function without paraplegia. Conclusion This awake rabbit model is highly reproducible and will be helpful in future studies of delayed paraplegia after spinal cord ischemia. The ambient temperature must be considered while using this model during investigation of therapeutic interventions.

Abstract Aiming at the problems of traditional ship cold chain monitoring systems being easily affected by environmental factors, difficult to achieve real‐time monitoring in the open sea without network signals, and low efficiency in transmitting Beidou short message data, a multi‐link compressed transmission ship cold chain monitoring system was designed by combining 5G technology, Beidou short message transmission technology, and multi‐protocol transmission technology. The system can adaptively switch the strength of wireless signals to ensure that information transmission is not lost. At the same time, a Beidou short message compressed transmission method was proposed to improve transmission efficiency. Test results show that the system can accurately complete data collection and processing, with small system errors, effectively improving the reliability and efficiency of the monitoring system, and has high application value.

Abstract In this study, selective adsorption experiments of Au were carried out using graphene oxide (GO) and thiourea-reduced GO (TU-rGO). Adsorption experiments using Au, Cu, Pb, and Zn were performed with GO and TU-rGO in order to selectively adsorb Au from simulated waste electric and electronic equipment leachate or printed circuit board wastewater. Optimal adsorption conditions were determined, adsorption isotherm models were fitted, and desorption and regeneration experiments were performed. Additionally, GO and TU-rGO were characterized to better understand the material and propose the adsorption and desorption mechanisms for TU-rGO. It was found that TU-rGO could selectively adsorb Au, achieving a high efficiency of 95‒99% at initial Au concentrations of 0.1‒10 mg L−1 with little to no adsorption of Cu, Pb, and Zn. Moreover, the desorption efficiency of Au by ammonium thiosulfate reached 94% with the adsorption efficiency of TU-rGO decreasing from 99 to 78% after five adsorption and desorption cycles. Isotherm adsorption experiments indicated that the Langmuir model better fitted the adsorption of Au than the Freundlich model. This implied that the adsorption process was mainly controlled by monolayer coverage, with a high Au adsorption capacity of approximately 833 mg g−1 for TU-rGO.

Summary: Autophagosome formation initiated on the endoplasmic reticulum (ER)-associated omegasome requires LC3. Translational regulation of LC3 biosynthesis is unexplored. Here we demonstrate that LC3 mRNA is recruited to omegasomes by directly binding to the ER transmembrane Sigma-1 receptor (S1R). Cell-based and in vitro reconstitution experiments show that S1R interacts with the 3′ UTR of LC3 mRNA and ribosomes to promote LC3 translation. Strikingly, the 3′ UTR of LC3 is also required for LC3 protein lipidation, thereby linking the mRNA-3′ UTR to LC3 function. An autophagy-defective S1R mutant responsible for amyotrophic lateral sclerosis cannot bind LC3 mRNA or induce LC3 translation. We propose a model wherein S1R de-represses LC3 mRNA via its 3′ UTR at the ER, enabling LC3 biosynthesis and lipidation. Because several other LC3-related proteins use the same mechanism, our data reveal a conserved pathway for localized translation essential for autophagosome biogenesis with insights illuminating the molecular basis of a neurodegenerative disease.

Southern blight is a common and devastating disease adversely affecting the yield and quality of cultivated Dendrobium officinale plants. The pathogens responsible for this disease are Sclerotium delphinii and S. rolfsii, although S. delphinii is more pathogenic than S. rolfsii. Plant endophytes are potential sources of biological control agents. In this study, 18 microbial strains were isolated from wild Dendrobium shixingense plants (e.g., Streptomyces, Bacillus, and Trichoderma), among which strain MEPF0303 significantly inhibited S. delphinii mycelial growth. Based on 16S rRNA gene sequence, this strain was identified as Streptomyces sp. In addition, the volatile organic compounds and cell-free supernatant of MEPF0303 had significant inhibitory effects on S. delphinii growth. Moreover, MEPF0303 effectively controlled southern blight of D. officinale. A total of 1,797 differentially expressed genes were identified by a comparative transcriptome analysis of S. delphinii mycelia treated with MEPF0303 and the untreated control mycelia. Strain MEPF0303 significantly altered the expression of genes related to cell membrane morphological development and integrity, pathogenicity, energy metabolism, and oxidative stress responses. Furthermore, the transcriptome sequencing results were validated by analyzing gene expression via a quantitative real-time polymerase chain reaction. The study results showed that MEPF0303 can inhibit the growth and pathogenicity of S. delphinii, with potential implications for the biological control of southern blight of D. officinale.

In this paper, the problem of stability of systems with a time delay is investigated. The time delay is assumed to vary in an interval, and its derivative is unknown. An augmented Lyapunov-Krasovskii functional (LKF) is constructed considering the time-varying delay fully. By augmenting several vectors and introducing some variable-augmented-based free-weighting matrices (FWMs), quadric terms, or higher-order terms of the time-varying delay do not appear in the derived stability conditions. Thus, the derived stability condition is presented as linear matrix inequalities (LMIs), which existing LMI tools can readily check. Finally, two examples, including a commonly used numerical example and the stability of load-frequency control (LFC) in single-area power systems, are provided to demonstrate the validity and merits of the proposed method.

The mitochondrial permeability transition pore is a nonspecific transmembrane channel. Inhibition of mitochondrial permeability transition pore opening has been shown to alleviate mitochondrial swelling, calcium overload, and axonal degeneration. Cyclophilin D is an important component of the mitochondrial permeability transition pore. Whether cyclophilin D participates in mitochondrial impairment and axonal injury after intracerebral hemorrhage is not clear. In this study, we established mouse models of intracerebral hemorrhage in vivo by injection of autologous blood and oxyhemoglobin into the striatum in Thy1-YFP mice, in which pyramidal neurons and axons express yellow fluorescent protein. We also simulated intracerebral hemorrhage in vitro in PC12 cells using oxyhemoglobin. We found that axonal degeneration in the early stage of intracerebral hemorrhage depended on mitochondrial swelling induced by cyclophilin D activation and mitochondrial permeability transition pore opening. We further investigated the mechanism underlying the role of cyclophilin D in mouse models and PC12 cell models of intracerebral hemorrhage. We found that both cyclosporin A inhibition and short hairpin RNA interference of cyclophilin D reduced mitochondrial permeability transition pore opening and mitochondrial injury. In addition, inhibition of cyclophilin D and mitochondrial permeability transition pore opening protected corticospinal tract integrity and alleviated motor dysfunction caused by intracerebral hemorrhage. Our findings suggest that cyclophilin D is used as a key mediator of axonal degeneration after intracerebral hemorrhage; inhibition of cyclophilin D expression can protect mitochondrial structure and function and further alleviate corticospinal tract injury and motor dysfunction after intracerebral hemorrhage. Our findings provide a therapeutic target for preventing axonal degeneration of white matter injury and subsequent functional impairment in central nervous diseases.

We build a model of the origin of physical constants, including masses and coupling constants. We consider the quantum correction of masses and coupling constants. Some exactly solved leading quantum corrections are given. In the model, the physical constant originates from a coupling between the matter field and a background field. We show that if such a background field as it should be in the real physical world is a quantum field, then the physical constant will have a space- and time-dependent quantum correction and will no longer be a constant. We build a scalar field model and a mechanics model. In these two models, we discuss the quantum correction of masses and coupling constants in the field framework and in the mechanics framework.

Three principal ER quality-control mechanisms, namely, the unfolded protein response, ER-associated degradation (ERAD), and ER-phagy are each important for the maintenance of ER homeostasis, yet how they are integrated to regulate ER homeostasis and organellar architecture in vivo is largely unclear. Here we report intricate crosstalk among the 3 pathways, centered around the SEL1L-HRD1 protein complex of ERAD, in the regulation of organellar organization in β cells. SEL1L-HRD1 ERAD deficiency in β cells triggers activation of autophagy, at least in part, via IRE1α (an endogenous ERAD substrate). In the absence of functional SEL1L-HRD1 ERAD, proinsulin is retained in the ER as high molecular weight conformers, which are subsequently cleared via ER-phagy. A combined loss of both SEL1L and autophagy in β cells leads to diabetes in mice shortly after weaning, with premature death by approximately 11 weeks of age, associated with marked ER retention of proinsulin and β cell loss. Using focused ion beam scanning electron microscopy powered by deep-learning automated image segmentation and 3D reconstruction, our data demonstrate a profound organellar restructuring with a massive expansion of ER volume and network in β cells lacking both SEL1L and autophagy. These data reveal at an unprecedented detail the intimate crosstalk among the 3 ER quality-control mechanisms in the dynamic regulation of organellar architecture and β cell function.

Abstract. Objective:. Blocking mechanistic target of rapamycin (mTOR) activation with mTOR inhibitors has promising therapeutic potential for keloids. However, the precise mechanism of mTOR inhibitors remains unclear. This study was aimed to investigate the role of autophagy machinery in the anti-keloid effects of mTOR inhibitors. Methods:. We first validated the biological effects induced by the mTOR inhibitors rapamycin (100 nmol/L) and KU-0063794 (5 μmol/L) on the proliferation, apoptosis, migration, and collagen synthesis of keloid fibroblasts (KFs) derived from Han Chinese persons through a Cell Counting Kit-8 assay, 5-Bromo-2’-deoxyuridine incorporation, Annexin V/propidium iodide staining, migration, and western blotting. To explore whether autophagy machinery is involved in the anti-keloid effects of mTOR inhibitors, we first blocked the autophagy activation induced by rapamycin and KU-0063794 with a pharmacological autophagy inhibitor (wortmannin) or by silencing the key autophagy gene (ATG5), and we then re-evaluated these biological effects on KFs. Results:. Blocking mTOR activation with either rapamycin or KU-0063794 completely inhibited proliferation, migration, and collagen synthesis of primary KFs but did not affect apoptosis. Incubating KFs with the autophagy inhibitor wortmannin or performing ATG5 silencing abrogated the subsequent activation of autophagic activity induced by rapamycin (rapamycin + E-64d + pepstatin vs. rapamycin + wortmannin + E-64d + pepstatin: 1.88 ± 0.38 vs. 1.02 ± 0.35, F = 6.86, P = 0.013), (non-sense control + rapamycin vs. ATG5 small interfering RNA + rapamycin: 1.46 ± 0.18 vs. 0.75 ± 0.20, respectively; F = 7.68, P = 0.01) or KU-0063794 (KU-0063794 + E-64d + pepstatin vs. KU-0063794 + wortmannin + E-64d + pepstatin: 1.65 ± 0.35 vs. 0.76 ± 0.17, F = 10.01, P = 0.004), (NC + KU-0063794 vs. ATG5 small interfering RNA + KU-0063794: 1.59 ± 0.50 vs. 0.77 ± 0.09, F = 5.93, P = 0.02) as evidenced by decreased accumulation of LC3-II. However, blockage of autophagy induction in mTOR inhibitor-treated KFs with both methods did not disturb their anti-keloid effects, such as inhibition of cell viability, cell migration, and collagen synthesis (P > 0.05 each). Conclusion:. Blocking mTOR activation with the mTOR inhibitors rapamycin and KU-0063794 showed anti-keloid effects in KFs. Restoration of autophagy inhibition by mTOR inhibitors does not contribute to their anti-keloid effects.

Abstract. Ultraviolet (UV) induced skin inflammation (UISI) is associated with many skin disorders. However, the mechanism by which UV causes skin inflammation remains unclear. Studies evaluating UISI in vivo have mainly been conducted using mouse models. Current investigations indicate that the classic inflammatory pathways involving nuclear factor kappa B and Toll-like receptor contribute to the regulation of UISI. However, more novel signaling factors have been identified as being involved in this process, including interleukin 22 receptor-α, cluster of differentiation 28 and cluster of differentiation 1d, serum amyloid A1, estrogen, melatonin, peroxisome proliferator activated receptors β/δ, isocitrate dehydrogenase 2, and transglutaminase 2. In addition, the gene mutation of fermitin family member 1 and selenium deficiency are reported to affect the phenotype of UISI. Although the actual roles of UISI in UV-related skin diseases need to be clarified, recent studies have reported the potent contribution of UISI to photocarcinogesis. To clarify the process and modulation of UISI, the special profiles of cytokines and inflammatory mediators and the core regulatory pathways should be identified clearly. These investigations would be promoted rapidly, accompanied by the conduction of high-quality clinical research on patients with UV-related skin disease and the construction of precise animal models of UISI.

Degradation of misfolded proteins from the endoplasmic reticulum (ER) is important for maintaining proper cellular protein homeostasis. Here, the authors discovered that the ER membrane protein PGRMC1 binds to misfolded prohormones for recruitment into the ER-phagy degradative pathway.

Abstract Background Cancer and ischemic stroke are two common diseases that threaten human health and have become the main causes of death in the world. It is estimated that one‐in‐ten patients with ischemic stroke have concomitant cancer, and this incidence is expected to increase as improvements in medical technology extends the life expectancy of cancer patients. Discussion Cancer‐related stroke (CRS) refers to unexplained ischemic stroke in patients with active cancer that cannot be explained by current stroke mechanisms. Available evidence suggests that CRS accounts for 5–10% of embolic stroke of undetermined source (ESUS). Although the incidence of CRS is gradually increasing, its underlying pathogenesis remains unclear. Also, there is no consensus on acute treatment and secondary prevention of stroke. Conclusion In this review, we retrospectively analyzed the incidence, mechanisms of CRS, its potential as a new stroke subtype, options for acute treatment, secondary prevention strategies, and disease progression, with the aim of attempting to explore personalized therapy strategies.

Yu-An Chen et al characterize the role of cell surface epitopes of the tumor suppressor protein WWOX in cell–cell recognition. The find that cells expressing functional WWOX, repel cancer cells which express dysfunctional WWOX or lack this protein, in a manner mediated by WWOX epitope 286-299.

Abstract Obesity increases the risk of developing cardiovascular disease and other metabolic diseases. We intended to compare three different anthropometric indicators of obesity, in predicting the incidence of cardiovascular events in Chinese type 2 diabetes. Beijing Community Diabetes Study was a prospective multi-center study conducted in Beijing community health centers. Type 2 diabetes patients from fourteen community health centers were enrolled at baseline. The primary endpoint was cardiovascular events. The upper quartile of neck circumference (NC) was set as greater NC. A total of 3299 diabetes patients were enrolled. In which, 941 (28.52%) had cardiovascular disease at baseline. Logistic analysis showed that central obesity (waist circumference (WC) above 90 cm in men and 85 cm in women) and greater NC were all related to baseline cardiovascular disease (adjusted OR = 1.49, and 1.55). After 10-year follow-up, 340 (10.31%) had cardiovascular events. Compared with patients without cardiovascular events, those having cardiovascular events had higher BMI, larger WC and NC. Cox regression analysis showed that greater WC and NC were all associated with the occurrence of cardiovascular events (adjusted HR = 1.41, and 1.38). A higher NC and WC might increase the risk of cardiovascular events by about 40% in type 2 diabetes patients in Beijing communities.

Glucose sensors based blood glucose detection are of great significance for the diagnosis and treatment of diabetes because diabetes has aroused wide concern in the world. In this study, bovine serum albumin (BSA) was used to cross-link glucose oxidase (GOD) on a glassy carbon electrode (GCE) modified by a composite of hydroxy fullerene (HFs) and multi-walled carbon nanotubes (MWCNTs) and protected with a glutaraldehyde (GLA)/Nafion (NF) composite membrane to prepare a novel glucose biosensor. The modified materials were analyzed by UV-visible spectroscopy (UV-vis), transmission electron microscopy (TEM), and cyclic voltammetry (CV). The prepared MWCNTs-HFs composite has excellent conductivity, the addition of BSA regulates MWCNTs-HFs hydrophobicity and biocompatibility, and better immobilizes GOD on MWCNTs-HFs. MWCNTs-BSA-HFs plays a synergistic role in the electrochemical response to glucose. The biosensor shows high sensitivity (167 μA·mM−1·cm−2), wide calibration range (0.01–3.5 mM), and low detection limit (17 μM). The apparent Michaelis–Menten constant Kmapp is 119 μM. Additionally, the proposed biosensor has good selectivity and excellent storage stability (120 days). The practicability of the biosensor was evaluated in real plasma samples, and the recovery rate was satisfactory.

Rutin is a natural antioxidant flavonoid compound with anti-inflammatory, antioxidant, and antiviral effects that is used to prepare drugs with wide application in clinical treatment. Therefore, the quantitative detection of rutin has important practical significance. In this work, a novel electrochemical sensor based on glassy carbon electrodes (GCEs) modified with sodium carboxymethylcellulose (CMC), multi-walled carbon nanotubes (MWCNTs), and 1-butyl-3-methylimid (ionic liquid, IL) was developed for the super-sensitive detection of the flavonoid rutin. The properties of these modified materials were analyzed by transmission electron microscope (TEM), cyclic voltammograms (CVs), and electrochemical-impedance spectroscopy (EIS). CMC was used to disperse MWCNTs to further enhance their hydrophilicity and biocompatibility. The modified MWCNTs improved the sensitivity of rutin detection. The square-wave voltammetry (SWV) technique showed that the linear range of rutin concentration determination was 0.01 μM to 1 µM and 1 µM to 10 µM. The minimum concentration detection of rutin was 0.83 nM and 6.6 nM, respectively. The proposed sensor presented good selectivity for rutin and successfully analyzed rutin content in the pharmaceutical rutin tablets. These results are consistent with those measured by ultra-high-performance liquid chromatography (UHPLC). Therefore, this sensor has latent application value in the analysis of rutin in food and drug tablets and nutraceutical samples.

Drug abuse has been and continues to be, a common social issue worldwide, yet the efficiency of widely adopted sweeping speech for anti-drug campaigns has proven inefficient. To provide students with a safe and efficient learning situation related to drug refusal skills, we used a novel approach rooted in a serious learning game and concept map during a brief extracurricular period to help students understand drugs and their negative effects. The proposed game-based situational learning system allowed all students to participate simultaneously and individually in multiple scenarios of drug temptation posed by peers and classmates to practice responding and refusing drugs in school and community settings. Moreover, to explore whether different personality traits (such as the Big Five personality traits) result in different anti-drug responses, we used a serious game to conduct an anti-drug experiment on 53 junior middle school students aged 13–15. Each participant’s decision-making process was recorded in the serious game as behavioral patterns for lag sequential analysis (LSA). The outcomes revealed seven behavioral patterns including differentiation (D), acceptance (A), effective (ER) and ineffective responses (IR), effective (ES) and ineffective solution-seeking (IS), and failure to refuse (F). The GSEQ (Generalized Sequential Querier) which is a computer program for analyzing sequential observational data was used. The results indicated the following: (1) Neuroticism was performed at a relatively low level under the guidance of a concept map. (2) “Neuroticism” was associated with the lowest risk of accepting drugs. (3) Students with “openness to experiences” were at high risk of accepting drugs. (4) Almost all personality behavioral transition diagrams showed that failure to refuse (F) drugs was followed by inefficient seeking of help (SI) and inefficient refusal (RI). These findings provide reference points for designing adaptive anti-drug education programs.

Pleurotus ostreatus (Jacq.) P. Kumm is cultivated worldwide, and its growth is seriously threatened by heat stress. Here, we performed a comprehensive analysis to investigate the influence of the phytohormone salicylic acid (SA) in P. ostreatus under HS. The results showed that the hyphal growth recovery rate and the antioxidant capacity of P. ostreatus increased with exogenous SA application (0.01 mmol/L and 0.05 mmol/L) after HS treatment. Metabolomic and transcriptomic analyses showed that SA application (0.05 mmol/L) weakened central carbon metabolism to allow cells to survive HS efficiently. In addition, SA shifted glycolysis to one-carbon metabolism to produce ROS scavengers (GSH and NADPH) and reduced ROS production by altering mitochondrial metabolism. SA also maintained nucleotide homeostasis, led to membrane lipid remodeling, activated the MAPK pathway, and promoted the synthesis of cell-wall components. This study provides a reference for further study of SA in microorganisms.

Theoretical predictions of carbon dioxide absorption flux were analyzed by developing one-dimensional mathematical modeling using the chemical absorption theory based on mass-transfer resistances in series. The CO2 absorption into monoethanolamine (MEA) solutions was treated as chemical absorption, accompanied by a large equilibrium constant. The experimental work of the CO2 absorption flux using MEA solution was conducted in double-unit flat-plate membrane contactors with embedded 3D turbulence promoters under various absorbent flow rates, CO2 feed flow rates, and inlet CO2 concentrations in the gas feed stream for both concurrent and countercurrent flow operations. A more compact double-unit module with embedded 3D turbulence promoters could increase the membrane stability to prevent flow-induced vibration and enhance the CO2 absorption rate by overwhelming the concentration polarization on the membrane surfaces. The measured absorption fluxes with a near pseudo-first-order reaction were in good agreement with the theoretical predictions for the CO2 absorption efficiency in aqueous MEA solutions, which was shown to be substantially larger than the physical absorption in water. By embedding 3D turbulence promoters in the MEA feed channel, the new design accomplishes a considerable CO2 absorption flux compared with an empty channel as well as the single unit module. This demonstrates the value and originality of the present study regarding the technical feasibility. The absorption flux enhancement for the double-unit module with embedded 3D turbulence promoters could provide a maximum relative increase of up to 40% due to the diminution in the concentration polarization effect. The correlated equation of the average Sherwood number was obtained numerically using the fourth Runge–Kutta method in a generalized and simplified expression to calculate the mass transfer coefficient of the CO2 absorption in the double-unit flat-plate membrane contactor with turbulence promoter channels.

Abstract Objectives Triple‐negative breast cancer (TNBC) is well known for its strong invasiveness, rapid recurrence and poor prognosis. Immunotherapy, including chimeric antigen receptor‐modified T (CAR‐T) cells, has emerged as a promising tool to treat TNBC. The identification of a specific target tumor antigen and the design of an effective CAR are among the many challenges of CAR‐T therapy. Methods We reported that epidermal growth factor receptor (EGFR) is highly expressed in TNBC and consequently designed an optimal third generation of CAR targeting EGFR. The efficacy of primary T lymphocytes infected with EGFR CAR lentivirus (EGFR CAR‐T) against TNBC was evaluated both in vitro and in vivo. The signalling pathways activated in tumor and EGFR CAR‐T cells were revealed by RNA sequencing analysis. Results Third‐generation EGFR CAR‐T cells exerted potent and specific suppression of TNBC cell growth in vitro, whereas limited cytotoxicity was observed towards normal breast epithelial cells or oestrogen receptor‐positive breast cancer cells. This capability was further demonstrated in vivo in a xenograft mouse model, with minimal off‐tumor cytotoxicity. Mechanistically, in vitro stimulation with TNBC cells induced the expansion of naïve‐associated EGFR CAR‐T cells and enhanced their persistence. Furthermore, EGFR CAR‐T cells activated the interferon γ, granzyme–perforin–PARP and Fas–FADD–caspase signalling pathways in TNBC cells. Conclusion We demonstrate that EGFR is a relevant immunotherapeutic target in TNBC, and EGFR CAR‐T exhibits potent and specific antitumor activity against TNBC, suggesting the potential of this third‐generation EGFR CAR‐T as an immunotherapy tool to treat TNBC in the clinic.

Gravitational waves are regarded as linear waves in the weak field approximation, which ignores the spacetime singularity. In this paper, we analyze singularities in exact gravitational wave solutions. We provide an exact general solution of the gravitational wave with cylindrical symmetry. The general solution includes some known cylindrical wave solutions as special cases. We show that there are two kinds of singularities in the cylindrical gravitational wave solution. The first kind of singularity corresponds to a singular source. The second kind of singularity corresponds to a resonance between different gravitational waves. When two gravitational waves coexist, the interference term in the source may vanish in the sense of time averaging.

The effects of antibacterial compounds produced by Saccharomyces cerevisiae in Koumiss on pathogenic Escherichia coli 08and its cell surface characteristics were investigated. S. cerevisiae isolated from Koumiss produced antibacterial compounds which were active against pathogenic E. coli O8 as determined by the Oxford cup method. The aqueous phases from S. cerevisiae at pH=2.0 (S2) and pH=8.0 (S8) were extracted and tested, respectively. The organic acids of S2 and S8 were determined by high performance liquid chromatography (HPLC), and the concentrations of killer toxins were determined by enhanced bicinchoninic acid (BCA) Protein Assay Kit. The minimum inhibition concentration (MIC) and the minimum bactericidal concentration (MBC) of S2 and S8 on E. coli O8 were determined by the broth microdilution method. The effects of S2 and S8 on the growth curve of E. coli O8were determined by turbidimetry, and the hydrophobicities of E. coli O8 cell surface were determined using the microbial adhesion to solvents method, the permeation of E. coli O8cell membrane were determined by the o-nitrophenyl-β-D-galactoside (ONPG) method. Aqueous phases at pH 2.0 and 8.0 had larger inhibition zones and then S2 and S8 were obtained by freeze-drying. The main component in S2 was citric acid and it was propanoic acid in S8. Other organic acids and killer toxins were also present. Both the MICs of S2 and S8 on E. coli O8 were 0.025 g mL−1, the MBCs were 0.100 and 0.200 g mL−1, respectively. The normal growth curve of E. coli O8was S-shaped, however, it changed after addition of S2 and S8. E. coli O8was the basic character, and had a relatively hydrophilic surface. The hydrophobicity of E. coli O8cell surface and the permeation of E. coli O8cell membrane were increased after adding S2 and S8. The present study showed that S2 and S8 inhibit the growth of pathogenic E. coli O8 and influence its cell surface characteristics.

Mutations in PLA2G6 gene cause PLA2G6-associated neurodegeneration, including recessive familial type 14 of Parkinson's disease (PARK14). Previously, we identified PARK14 patients with compound heterozygous c.991G > T/c.1077G > A (p.D331Y/p.M358IfsX) mutations. The c.1077G > A mutation led to a four base-pairs deletion and frameshift mutation (p.M358IfsX) of PLA2G6 mRNA. We established induced pluripotent stem cells (iPSCs) from peripheral blood mononuclear cells of a female patient with compound heterozygous c.991G > T/c.1077G > A (p.D331Y/ p.M358IfsX) mutations by using Sendai-virus delivery system. The iPSCs exhibited pluripotency and in vivo differentiation potential. The iPSCs can be used for studying the molecular pathogenic mechanism of PARK14.

MicroRNAs (miRs) downregulate or upregulate the mRNA level by binding to the 3′-untranslated region (3′UTR) of target gene. Dysregulated miR levels can be used as biomarkers of Parkinson’s disease (PD) and could participate in the etiology of PD. In the present study, 45 brain-enriched miRs were evaluated in serum samples from 50 normal subjects and 50 sporadic PD patients. The level of miR-204-5p was upregulated in serum samples from PD patients. An upregulated level of miR-204-5p was also observed in the serum and substantia nigra (SN) of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of PD. Expression of miR-204-5p increased the level of α-synuclein (α-Syn), phosphorylated (phospho)-α-Syn, tau, or phospho-tau protein and resulted in the activation of endoplasmic reticulum (ER) stress in SH-SY5Y dopaminergic cells. Expression of miR-204-5p caused autophagy impairment and activation of c-Jun N-terminal kinase (JNK)-mediated apoptotic cascade in SH-SY5Y dopaminergic cells. Our study using the bioinformatic method and dual-luciferase reporter analysis suggests that miR-204-5p positively regulates mRNA expression of dual-specificity tyrosine phosphorylation regulated kinase 1A (DYRK1A) by directly interacting with 3′UTR of DYRK1A. The mRNA and protein levels of DYRK1A were increased in SH-SY5Y dopaminergic cells expressing miR-204-5p and SN of MPTP-induced PD mouse model. Knockdown of DYRK1A expression or treatment of the DYRK1A inhibitor harmine attenuated miR-204-5p-induced increase in protein expression of phospho-α-Syn or phospho-tau, ER stress, autophagy impairment, and activation of JNK-mediated apoptotic pathway in SH-SY5Y dopaminergic cells or primary cultured dopaminergic neurons. Our results suggest that upregulated expression of miR-204-5p leads to the death of dopaminergic cells by targeting DYRK1A-mediated ER stress and apoptotic signaling cascade.

Summary: Although flaviviruses co-opt the function of the host endoplasmic reticulum (ER) membrane protein complex (EMC) during infection, a mechanistic explanation for this observation remains unclear. Here, we show that the EMC promotes biogenesis of dengue virus (DENV) and Zika virus (ZIKV) non-structural multi-pass transmembrane proteins NS4A and NS4B, which are necessary for viral replication. The EMC binds to NS4B and colocalizes with the DENV replication organelle. Mapping analysis reveals that the two N-terminal marginally hydrophobic domains of NS4B confer EMC dependency. Furthermore, altering the hydrophobicity of these two marginally hydrophobic domains relieves NS4B’s EMC dependency. We demonstrate that NS4B biogenesis, but not its stability, is reduced in EMC-depleted cells. Our data suggest that the EMC acts as a multi-pass transmembrane chaperone required for expression of at least two virally encoded proteins essential for flavivirus infection and point to a shared vulnerability during the viral life cycle that could be exploited for antiviral therapy. : Multiple genetic screens have identified the ER membrane protein complex (EMC) as essential for infection by dengue and Zika flaviviruses. Lin et al. demonstrate that efficient biogenesis of the viral non-structural proteins NS4A and NS4B requires the EMC. Keywords: flavivirus, dengue virus, Zika virus, endoplasmic reticulum, ER membrane protein complex

During entry, a virus must be transported through the endomembrane system of the host cell, penetrate a cellular membrane, and undergo capsid disassembly, to reach the cytosol and often the nucleus in order to cause infection. To do so requires the virus to coordinately exploit the action of cellular membrane transport, penetration, and disassembly machineries. How this is accomplished remains enigmatic for many viruses, especially for viruses belonging to the nonenveloped virus family. In this review, we present the current model describing infectious entry of the nonenveloped polyomavirus (PyV) SV40. Insights from SV40 entry are likely to provide strategies to combat PyV-induced diseases, and to illuminate cellular trafficking, membrane transport, and disassembly mechanisms.