Synthesis and Luminescence of POSS-Containing Perylene Bisimide-Bridged Amphiphilic Polymers

A novel well-defined amphiphilic fluorescent polymer containing asymmetric perylene bisimide was designed and synthesized by combining reaction of perylene anhydride with amino functional polyhedral oligomeric silsesquioxane (POSS) and atom transfer radical polymerization (ATRP) of <i>N</i>-isopropylacrylamide (NIPAM). All the intermediate and final products were characterized by NMR, Fourier transform infrared spectroscopy (FT-IR), elemental analyses, and gel permeation chromatograph (GPC). Self-assembly of the amphiphilic polymers was investigated in aqueous solution and POSS-containing hybrid nanoparticles were obtained and characterized by dynamic laser light scattering (DLS) and transmission electron microscopy (TEM). The novel hybrid nanoparticles exhibit attractive high red fluorescence at 645 nm due to the significant effect of the bulky POSS moieties. Moreover, based on the thermoresponsive PNIPAM coronas, the fluorescence intensity of the self-assembled hybrid nanoparticles can be further enhanced and tuned by changing temperature

Immunofluorescence identification of human Tenon fibroblasts.

<p>Cells were identified by immunofluorescence with vimentin and cytokeratin. Propidium iodide was used as a nuclear counter stain. (<b>A</b>) Immunofluorescence double-labelling of vimentin (green) and the nuclear (red). (<b>B</b>) Cells did not express cytokeratin.</p

Rosiglitazone and SB203580 attenuated activation of fibroblast motility induced by TGF-β1.

<p>Stimulation with TGF-β1 significantly improved the fibroblast motility compared with control group (<i>p</i><0.05). However, both rosiglitazone and SB203580 suppressed the enhancing effect of TGF-β1 on cell migration. There is no significant difference between rosiglitazone group and SB203580 group (<i>p</i>>0.05). Cells were seeded in six-well plates, and scratch wounds were performed. After washing away suspended particles, we preincubated the cells with rosiglitazone for 2 h or SB203580 for 1 h in serum-starved medium. Then, the cells were stimulated with 5 ng/mL TGF-β1 for another 24 h in serum-free medium in the presence of rosiglitazone or SB203580. Width of wound was measured at six different sites and averaged in each sample. The width in each group was expressed as a percentage of the original measurement before stimulation, and the original width was presented as 100%. Each bar (mean) is the average of repetitions of three repeated experiments, and the error bars shown the standard deviation. Asterisk indicates p<0.05 compared with control group; Dagger indicates p<0.05 compared with TGF-β1 group.</p

Rosiglitazone attenuated activation of the p38 signal pathway induced by TGF-β1.

<p>(<b>A</b>) TGF-β1 increased the expression of phosphorylated p38 in cultured fibroblasts. Cells were were stimulated with 5 ng/mL TGF-β1 under serum-free conditions. The phosphorylated p38 MAPK level was measured at various time points from 15 to 240 min. Phosphorylated p38 as measured by western blot increased after 15 min and peaked at 30 min. Total p38 expression was unchanged before and after stimulation with TGF-β1. This experiment was repeated 3 times with similar results. (<b>B</b>) Rosiglitazone exhibited no significant effect on the phosphorylation of p38 in unstimulated cells (<i>p</i>>0.05). However, the addition of rosiglitazone significantly suppressed p38 pathway activation in the cells stimulated with TGF-β1. Each bar (mean) is the average of repetitions of three repeated experiments, and the error bars shown the standard deviation. Asterisk indicates p<0.05 compared with control group; Dagger indicates p<0.05 compared with TGF-β1 group.</p

Rosiglitazone and SB203580 attenuated fibroblast proliferation induced by TGF-β1.

<p>(<b>A</b>) Cell Counting Kit-8 assays showed that treatment with TGF-β1 significantly increased the proliferation of fibroblasts after 24 h (<i>p</i><0.05). While the TGF-β1-induced proliferation was inhibited by SB203580 in a dose-dependent manner (<i>p</i><0.05). There is no significant differences among 10, 20 and 50 µmol/L (<i>p</i>>0.05). Cells were treated with TGF-β1 in the presence or absence of p38 specific inhibitor SB20380 at concentrations of 1, 10, 20 and 50 µmol/L. (<b>B</b>) SB203580 (10 µmol/L) alone exhibited no significant effect on fibroblast viability (<i>p</i>>0.05). The inhibitory effect of SB203580 on TGF-β1 stimulated cell proliferation was weaker than the effect of rosiglitazone (<i>p</i><0.05). Each bar (mean) is the average of repetitions of three repeated experiments, and the error bars shown the standard deviation. Asterisk indicates p<0.05 compared with control group; Dagger indicates p<0.05 compared with TGF-β1 group.</p

Recruitment of α-SMA in the cytoplasm of fibroblasts.

<p>Rosiglitazone and SB203580 attenuated the TGF-β1–induced upregulation of α-SMA: (<b>A</b>) vehicle control, (<b>B</b>) vehicle and TGF-β1, (<b>C</b>) rosiglitazone and TGF-β1, and (<b>D</b>) SB203580 and TGF-β1. Fibroblasts were preincubated with rosiglitazone or SB203580 in serum-starved medium. Then, the cells were incubated with 5 ng/mL TGF-β1 in the presence of rosiglitazone or SB203580 for another 48 h. Cells analyzed by immunofluorescence with α-SMA (green) as a representative fibrosis marker and propidium iodide to stain nuclear (red).</p

Rosiglitazone and SB203580 suppressed the mRNA expression of α-SMA, Col I, and CTGF.

<p>Fibroblasts were treated with rosiglitazone for 2 h or SB203580 for 1 h. Next, the cells were incubated with 5 ng/mL TGF-β1 in the presence of rosiglitazone or SB203580 for 12 h (for CTGF mRNA analysis) or 24 h (for α-SMA and Col I mRNA analysis). Fibroblasts treated with vehicle control expressed little α-SMA, Col I and CTGF, increased α-SMA, Col I and CTGF mRNA expression was observed in fibroblasts treated with TGF-β1 (<i>p</i><0.05, respectively). Furthermore, additional SB203580 treatments significantly decreased the TGF-β1–induced increase in α-SMA mRNA expression (<i>p</i><0.05). The expression of α-SMA in cells pretreated with rosiglitazone was lower than that in cells pretreated with SB203580 (<i>p</i><0.05). Cells were treated with vehicle, rosiglitazone, SB203580, and TGF-β1. Next, the cells were incubated with 5 ng/mL TGF-β1 in the presence of rosiglitazone or SB203580. α-SMA mRNA abundance was analyzed qPCR, and the housekeeping gene GAPDH transcript was used for normalization. Each bar (mean) is the average of repetitions of three repeated experiments, and the error bars shown the standard deviation. Asterisk indicates p<0.05 compared with control group; Dagger indicates p<0.05 compared with TGF-β1 group.</p

Data_Sheet_1_The Network of Interactions Among Porcine Reproductive and Respiratory Syndrome Virus Non-structural Proteins.docx

<p>The RNA synthesis of porcine reproductive and respiratory syndrome virus (PRRSV), a positive-strand RNA virus, is compartmentalized in virus-induced double-membrane vesicles where viral proteins and some cellular proteins assemble into replication and transcription complexes (RTCs). The viral replicase proteins are the major components of the RTCs but the physical associations among these non-structural proteins (nsps) remain elusive. In this study, we investigated the potential interactions between PRRSV nsps by yeast two-hybrid (Y2H), bimolecular fluorescence complementation (BiFC) and pull-down assays. Our analyses revealed a complex network of interactions involving most of PRRSV nsps. Among them, nsp9 and nsp12 were identified as the hubs of the nsp interactome; transmembrane proteins nsp2 and nsp5 both interacted with nsp3, indicating that the three membrane-bound proteins might bind together to form the scaffold to support the association of RTCs with the intracellular membrane. The PRRSV nsp interactions identified in this study may provide valuable clues for future researches on the RTC formation and function.</p

Table_2_A cross-sectional survey of medical and other groups’ awareness, perceptions, and willingness to use e-cigarettes during the COVID-19 pandemic.docx

BackgroundIn China, people’s perceptions towards electronic cigarettes during Corona Virus Disease 2019 (COVID-19) pandemic compared with pre-pandemic conditions have not been explored. Exploring the perceptions of medical workers regarding e-cigarettes is crucial, as they serve as a trusted source of information and providers of smoking cessation counselling for smokers. This cross-sectional study was designed to explore the awareness and perceptions of e-cigarettes among Chinese medical and other groups in the context of the COVID-19 pandemic.MethodsA cross-sectional survey was performed using an online, anonymous, and self-administered questionnaire. The questionnaire contained sections for collecting participants’ general information and Likert scale questions regarding smoking status, perceptions of e-smoking, attitude, and willingness to use e-cigarettes. The respondents included medical students, clinical doctors, and other occupations. Data analysis was performed using tools such as descriptive analysis, binary logistic regression, and multivariate regression.ResultsA total of 952 people completed the questionnaire, and 96.54% of them reported to have heard about e-cigarettes. The most common source of information about e-cigarettes was advertising. Notably, 28 of the 116 smokers reported that they had used e-cigarettes. Independent-samples T-tests results showed that medical groups believed e-cigarettes contained tar (p = 0.03). Most of the medical and non-medical participants maintained neutral attitudes towards e-cigarette policies (38.3%) and prices (49.2%) but their views were significantly different (p  40, OR = 9.794, 95CI% = 1.683–56.989, p = 0.011) and the older adult used fewer than the young (18–40, OR = 4.797, 95CI% = 0.930–24.744, p = 0.061; > 40, OR = 9.794, 95CI% = 1.683–56.989, p = 0.011).ConclusionThis study found that individuals tend to hold negative attitudes towards the awareness, perceptions, and willingness to use e-cigarettes. Medical groups are less likely to use e-cigarettes, but misperceptions are still prevalent among them. This calls for additional training for such medical personnel to improve their capacity to provide necessary counselling to smokers. E-cigarettes advertisements were the main source of information for young individuals to learn about e-cigarettes, and hence measures should be taken to restrict exposure of young individuals to e-cigarettes.</p

Table_1_A cross-sectional survey of medical and other groups’ awareness, perceptions, and willingness to use e-cigarettes during the COVID-19 pandemic.docx

BackgroundIn China, people’s perceptions towards electronic cigarettes during Corona Virus Disease 2019 (COVID-19) pandemic compared with pre-pandemic conditions have not been explored. Exploring the perceptions of medical workers regarding e-cigarettes is crucial, as they serve as a trusted source of information and providers of smoking cessation counselling for smokers. This cross-sectional study was designed to explore the awareness and perceptions of e-cigarettes among Chinese medical and other groups in the context of the COVID-19 pandemic.MethodsA cross-sectional survey was performed using an online, anonymous, and self-administered questionnaire. The questionnaire contained sections for collecting participants’ general information and Likert scale questions regarding smoking status, perceptions of e-smoking, attitude, and willingness to use e-cigarettes. The respondents included medical students, clinical doctors, and other occupations. Data analysis was performed using tools such as descriptive analysis, binary logistic regression, and multivariate regression.ResultsA total of 952 people completed the questionnaire, and 96.54% of them reported to have heard about e-cigarettes. The most common source of information about e-cigarettes was advertising. Notably, 28 of the 116 smokers reported that they had used e-cigarettes. Independent-samples T-tests results showed that medical groups believed e-cigarettes contained tar (p = 0.03). Most of the medical and non-medical participants maintained neutral attitudes towards e-cigarette policies (38.3%) and prices (49.2%) but their views were significantly different (p  40, OR = 9.794, 95CI% = 1.683–56.989, p = 0.011) and the older adult used fewer than the young (18–40, OR = 4.797, 95CI% = 0.930–24.744, p = 0.061; > 40, OR = 9.794, 95CI% = 1.683–56.989, p = 0.011).ConclusionThis study found that individuals tend to hold negative attitudes towards the awareness, perceptions, and willingness to use e-cigarettes. Medical groups are less likely to use e-cigarettes, but misperceptions are still prevalent among them. This calls for additional training for such medical personnel to improve their capacity to provide necessary counselling to smokers. E-cigarettes advertisements were the main source of information for young individuals to learn about e-cigarettes, and hence measures should be taken to restrict exposure of young individuals to e-cigarettes.</p