93 research outputs found

    Droplet behavior of non-equilibrium condensation in the supersonic separator

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    Optimization of production of PLA microbubble ultrasound contrast agents for Hydroxycamptothecin delivery

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    In this paper, ultrasound contrast agents based on a high molecular polymer poly lactic acid (PLA) and loaded with Hydroxycamptothecin (HCPT) were prepared by combining ultrasound method and a Shirasu Porous Glass (SPG) membrane emulsification technique. A special focus was on the optimization of production of RCPT-PLA microbubbles. Different factors, such as the power and the time of ultrasonic action, the ratio of inner aqueous phase against outer oil phase, and the concentration of PLA were evaluated, and the average size of HCPT-PLA microbubbles, the drug carrying efficiency, as well as the acoustically-triggered drug release at 3kHz ultrasound were determined. The study showed that the HCPT-PLA microbubbles prepared using our optimized conditions, were sphere-like in shape with a mean diameter of 1-7 mu m. The drug loading efficiency reached up to 56.48%. In vitro, the drug release of HCPT-PLA microbubbles increased significantly at 3kHz ultrasound for 30s compared with that of ultrasound free condition. In conclusion, the HCPT-PLA microbubbles has the characteristics desirable for an intravenously administered ultrasound contrast agent for further clinical use

    Construction of an Integrated mCherry Red Fluorescent Protein Expression System for Labeling and Tracing in Lactiplantibacillus plantarum WCFS1

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    Thorough intestinal adhesion and colonization greatly promote the probiotic properties of lactic acid bacteria (LAB). Labeling and tracing with fluorescent proteins are effective and reliable for studying the in vivo physiological activities of LAB including localization, adhesion, and colonization. Lactiplantibacillus plantarum WCFS1 was successfully traced with a red fluorescent protein (RFP), which was expressed by the bacteria-carrying recombinant plasmids. In this study, we aimed to construct a stable RFP mCherry expression system, whose encoding gene was integrated into the bacterial chromosome via double-crossed homologous recombination, and use it for labeling WCFS1 with the goal of avoiding the potential loss of non-chromosomal plasmids along with intestinal growth. First, the constitutive expression of the mCherry protein was improved after adjusting the length of the spacer between the promoter and the gene start codon. Then, the optimized mCherry gene expression cassette was integrated into the chromosome of WCFS1. The resulting strain had normal unimpaired growth and strong fluorescent signals, even after 100 generations, indicating its stability. Furthermore, quantitative polymerase chain reaction (PCR) results revealed a strong positive correlation between the fluorescence intensity of the strain and the number of viable cells, demonstrating its potential usage for the quantification of in vivo WCFS1 cells. Finally, the increased adhesion ability of WCFS1 due to the recombinant expression of the bsh gene was visualized and evaluated using fluorescence intensity, the results of which were consistent with those obtained using the previously established quantification methods. These results suggest that the chromosomal-integrated mCherry labeling system can be extensively used to examine the distribution, colonization, and survival of LAB in vivo in order to determine the mechanism of its probiotic function

    Biological Characteristics of Severe Combined Immunodeficient Mice Produced by CRISPR/Cas9-Mediated Rag2 and IL2rg Mutation

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    Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas)9 is a novel and convenient gene editing system that can be used to construct genetically modified animals. Recombination activating gene 2 (Rag2) is a core component that is involved in the initiation of V(D)J recombination during T- and B-cells maturation. Separately, the interleukin-2 receptor gamma chain gene (IL2rg) encoded the protein-regulated activity of natural killer (NK) cells and shared common receptors of some cytokines. Rag2 and IL2rg mutations cause immune system disorders associated with T-, B-, and NK cell function and some cytokine activities. In the present study, 2 single-guide RNAs (sgRNAs) targeted on Rag2 and IL2rg genes were microinjected into the zygotes of BALB/c mice with Cas9 messenger RNA (mRNA) to create Rag2/IL2rg-/- double knockout mice, and the biological characteristics of the mutated mice were subsequently analyzed. The results showed that CRISPR/Cas9-induced indel mutation displaced the frameshift of Rag2 and IL2rg genes, resulting in a decrease in the number of T-, B-, and NK cells and the destruction of immune-related tissues like the thymus and spleen. Mycobacterium tuberculosis 85B antigen could not induce cellular and humoral immune response in mice. However, this aberrant immune activity compromised the growth of several tumor heterogenous grafts in the mutated mice, including orthotopic and subcutaneous transplantation tumors. Thus, Rag2/IL2rg-/- knockout mice possessed features of severe combined immunodeficiency (SCID), which is an ideal model for human xenograft

    Muscle and Heart Function Restoration in a Limb Girdle Muscular Dystrophy 2I (LGMD2I) Mouse Model by Systemic FKRP Gene Delivery

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    Mutations in fukutin-related protein (FKRP) gene cause a wide spectrum of disease phenotypes including the mild limb-girdle muscular dystrophy 2I (LGMD2I), the severe Walker-Warburg syndrome, and muscle-eye-brain disease. FKRP deficiency results in α-dystroglycan (α-DG) hypoglycosylation in the muscle and heart, which is a biochemical hallmark of dystroglycanopathies. To study gene replacement therapy, we generated and characterized a new mouse model of LGMD2I harboring the human mutation leucine 276 to isoleucine (L276I) in the mouse alleles. The homozygous knock-in mice (L276IKI) mimic the classic late onset phenotype of LGMD2I in both skeletal and cardiac muscles. Systemic delivery of human FKRP gene by AAV9 vector in the L276IKI mice, at either neonatal age or at the age of 9 months, rendered body wide FKRP expression and restored glycosylation of α-DG in both skeletal and cardiac muscles. FKRP gene therapy ameliorated dystrophic pathology and cardiomyopathy such as muscle degeneration, fibrosis, and myofiber membrane leakage, resulting in restoration of muscle and heart contractile functions. Thus, these results demonstrated that the treatment based on FKRP gene replacement was effective

    The effect of inhibitory control and language proficiency on intra-sentential switching costs in reading comprehension

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    Based on the inhibition account and activation account, inhibitory control and language proficiency play big roles on language control mechanisms underlying language switching respectively. But it is still debatable whether inhibitory control and language proficiency play roles in sentential context during bilingual language comprehension, and how and when they work if they both do. The present study examined the specific roles of language proficiency and inhibitory control on modulating intra-sentential switching costs in Chinese-English bilinguals using self-paced reading task. Results indicated that language proficiency and switching direction modulated intra-sentential switching costs significantly. Switching costs were larger when switching into L1 than into L2 for the first two code-switched words due to inhibitory effect, but more costly switching into L2 than into L1 since the third code-switched words owing to the effect of relative language proficiency. During bilingual language processing, inhibition and activation of languages shaped a dynamic interplay of complementary language control processes. Specifically, inhibitory control and language proficiency work as a dynamic continuum in language switching at sentence level. Switching into L1 requires the release of inhibition to L1, but switching into L2 asks the activation and access of L2. The interaction effects of language proficiency and inhibitory control influenced the exertion and components of inhibition. This study reconciled the independent roles of language proficiency and inhibitory control on language switching and constructed a new continuum based on language proficiency

    A new active learning method based on the learning function U of the AK-MCS reliability analysis method

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    In recent years, reliability analysis methods based on the Kriging surrogate model have often been employed to obtain accurate failure probabilities of problems since the Kriging model can be used to provide predictions of the performance function at sample points and the corresponding variance of these predictions. Several learning functions have been explored to update the design of experiments and to complete the iterative process. However, it is still not easy to reduce the number of times the performance function or finite element model (FEM) is called for problems using the Kriging model. In this paper, a new active learning method based on a widely used learning function U is proposed to improve the speed of convergence of the AK-MCS method for problems with a connected domain of failure. Then, three academic examples and one three-unequal-span continuous girder with an implicit performance function are used to verify the accuracy and validity of the AK-MCS method based on the proposed learning method. Comparisons with AK-MCS based on the learning function U and MCS indicate that AK-MCS based on the proposed learning method requires calling the performance function or FEM fewer times than required by AK-MCS based on the learning function U to obtain accurate failure probabilities of the four examples, especially for six-dimensional problems. (C) 2017 Elsevier Ltd. All rights reserved

    Long-term high temperature stress decreases the photosynthetic capacity and induces irreversible damage in chrysanthemum seedlings

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    To study the effects of long-term and short-term high temperature stress and recovery on the physiological functions and appearance quality of chrysanthemums, a controlled experiment with chrysanthemums was conducted. The treatments were 25 °C for 3 days (T1D3), 25 °C for 9 days (T1D9), 41 °C for 3 days (T2D3) and 41°C for 9 days (T2D9). The results indicated that there is no significant difference between the T1D3 and T1D9 groups. Conversely, the total chlorophyll content (Chl), net photosynthetic rate (PN), and maximum quantum yield of Photosystem II (PSII) (FV/FM) under T2D3 and T2D9 decreased by 27.07%, 43.30%, 5.62%, and 44.85%, 68.22%, 8.29%, respectively. The JIP-test results showed that the T2D9-stressed plants had a lower efficiency and functional antenna size, and a higher activity of the reaction centre than T2D3. The contents of malondialdehyde, soluble protein and proline increased by 3.67 nmol/g FM, 298.75 μg/g, and 192.99 μg/g, and the antioxidant enzymes activities were inhibited significantly under T2D9. After the stress was relieved, Chl, PN, and FV/FM under T2D3 recovered to the same level as T1D3, while T2D9 did not. Furthermore, the diameter of the flowers in T2D3 showed no significant difference with the chrysanthemums under T1D3. However, the plants in T2D9 recovered poorly. Both the diameter of the flowers and the anthocyanin under T2D9 reduced significantly comparing with T1D9, indicating that the damage in the chrysanthemum seedlings caused by long-term high temperature was irreversible
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