C/N-Dependent Element Bioconversion Efficiency and Antimicrobial Protein Expression in Food Waste Treatment by Black Soldier Fly Larvae.

The black soldier fly (BSF), Hermetia illucens, has emerged as a promising species for waste bioconversion and source of antimicrobial proteins (AMPs). However, there is a scarcity of research on the element transformation efficiency and molecular characterization of AMPs derived from waste management. Here, food waste treatment was performed using BSF larvae (BSFL) in a C/N ratio of 21:1−10:1, with a focus on the C/N-dependent element bioconversion, AMP antimicrobial activity, and transcriptome profiling. The C-larvae transformation rates were found to be similar among C/Ns (27.0−35.5%, p = 0.109), while the N-larvae rates were different (p = 0.001), with C/N 21:1−16:1 (63.5−75.0%) being higher than C/N 14:1−10:1 (35.0−45.7%). The C/N ratio did not alter the antimicrobial spectrum of AMPs, but did affect the activities, with C/N 21:1 being significantly lower than C/N 18:1−10:1. The lysozyme genes were found to be significantly more highly expressed than the cecropin, defensin, and attacin genes in the AMP gene family. Out of 51 lysozyme genes, C/N 18:1 and C/N 16:1 up-regulated (p < 0.05) 14 and 12 genes compared with C/N 21:1, respectively, corresponding to the higher activity of AMPs. Overall, the element bioconversion efficiency and AMP expression can be enhanced through C/N ratio manipulation, and the C/N-dependent transcriptome regulation is the driving force of the AMP difference.

Anti-Inflammatory Effect of Dimethyl Fumarate Associates with the Inhibition of Thioredoxin Reductase 1 in RAW 264.7 Cells.

Macrophages secrete a variety of pro-inflammatory cytokines in response to pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs) but abnormal release of cytokines unfortunately promotes cytokine storms. Dimethyl fumarate (DMF), an FDA-approved drug for multiple sclerosis (MS) treatment, has been found as an effective therapeutic agent for resolution. In this study, the anti-inflammatory effect of DMF was found to correlate to selenoprotein thioredoxin reductase 1 (TXNRD1). DMF irreversibly modified the Sec498 residue and C-terminal catalytic cysteine residues of TXNRD1 in a time- and dose-dependent manner. In LPS-stimulated RAW 264.7 cells, cellular TXNRD activity was increased through up-regulation of the protein level and DMF inhibited TXNRD activity and the nitric oxide (NO) production of RAW 264.7 cells. Meanwhile, the inhibition of TXNRD1 by DMF would contribute to the redox regulation of inflammation and promote the nuclear factor erythroid 2-related factor 2 (NRF2) activation. Notably, inhibition of cellular TXNRD1 by auranofin or TRi-1 showed anti-inflammatory effect in RAW 264.7 cells. This finding demonstrated that targeting TXNRD1 is a potential mechanism of using immunometabolites for dousing inflammation in response to pathogens and highlights the potential of TXNRD1 inhibitors in immune regulation.

Carboxymethyl Chitosan and Gelatin Hydrogel Scaffolds Incorporated with Conductive PEDOT Nanoparticles for Improved Neural Stem Cell Proliferation and Neuronal Differentiation.

Tissue engineering scaffolds provide biological and physiochemical cures to guide tissue recovery, and electrical signals through the electroactive materials possess tremendous potential to modulate the cell fate. In this study, a novel electroactive hydrogel scaffold was fabricated by assembling poly(3,4-ethylenedioxythiophene) (PEDOT) nanoparticles on a carboxymethyl chitosan/gelatin (CMCS/Gel) composite hydrogel surface via in situ chemical polymerization. The chemical structure, morphology, conductivity, porosity, swelling rate, in vitro biodegradation, and mechanical properties of the prepared hydrogel samples were characterized. The adhesion, proliferation, and differentiation of neural stem cells (NSCs) on conductive hydrogels were investigated. The CMCS/Gel-PEDOT hydrogels exhibited high porosity, excellent water absorption, improved thermal stability, and adequate biodegradability. Importantly, the mechanical properties of the prepared hydrogels were similar to those of brain tissue, with electrical conductivity up to (1.52 ± 0.15) × 10-3 S/cm. Compared to the CMCS/Gel hydrogel, the incorporation of PEDOT nanoparticles significantly improved the adhesion of NSCs, and supported long-term cell growth and proliferation in a three-dimensional (3D) microenvironment. In addition, under the differentiation condition, the conductive hydrogel also significantly enhanced neuronal differentiation with the up-regulation of ß-tubulin III expression. These results suggest that CMCS/Gel-PEDOT hydrogels may be an attractive conductive substrate for further studies on neural tissue repair and regeneration.

An Organic Solvent-Free Method for the Extraction of Ellagic Acid Compounds from Raspberry Wine Pomace with Assistance of Sodium Bicarbonate.

Industrial processing of raspberry juice and wine generates considerable byproducts of raspberry pomace. Ellagic acids/ellagitannins, being characterized by their antioxidant and antiproliferation properties, constitute the majority of polyphenolics in the pomace and are valuable for recovery. In the present study, we developed a novel procedure with sodium bicarbonate assisted extraction (SBAE) to recover ellagic acid from raspberry wine pomace. Key parameters in the procedure, i.e., sodium bicarbonate concentration, temperature, time and solid/liquid (S/L) ratio, were investigated by single factor analysis and optimized subsequently by Response Surface Methodology (RSM). Optimal parameters for the SBAE method here were found to be 1.2% (w/v) NaHCO3, 1:93 (w/v) S/L ratio, 22 min and 100 °C. Under these conditions, the ellagic acid yield was 6.30 ± 0.92 mg/g pomace with an antioxidant activity of 79.0 ± 0.96 µmol Trolox eq/g pomace (DPPH assay), which are 2.37 and 1.32 times the values obtained by extraction with methanol-acetone-water solvent, respectively. The considerable improvement in ellagic acid extraction efficiency could be highly attributed to the reactions of lipid saponification and ellagitannin hydrolysis resulted from sodium bicarbonates. The present study has established an organic solvent-free method for the extraction of ellagic acid from raspberry wine pomace, which is feasible and practical in nutraceutical applications.

Electrical stimulation enhances the neuronal differentiation of neural stem cells in three-dimensional conductive scaffolds through the voltage-gated calcium ion channel.

Electrical stimulation (ES) or electroconductive scaffold has been proved to have the positive effects on the behavior of neural stem cells (NSCs). We previously developed a novel three-dimensional conductive composite scaffold of poly (3, 4-ethylenedioxythiophen)/chitosan/gelatin (PEDOT/Cs/Gel) for neural tissue engineering. In the present study, we further studied the effect of three-dimensional conductive scaffolds combined with ES on the neuronal differentiation of NSCs. The sandwiched ES device was designed to apply single-phase pulse voltage on NSCs cultured in conductive scaffold for 7 days (4 h/day). Proliferation and differentiation related proteins and genes were analyzed by immunofluorescence staining and RT-qPCR. The role of voltage-gated ion channels (VGICs) in regulating NSCs' neuronal differentiation by ES was investigated in presence of ion channels blockers. The results of protein and gene expression indicated that ES not only promoted the proliferation of NSCs cultured in the conductive scaffold, but also enhanced the differentiation of NSCs into neurons. Especially, the voltage-gated calcium channel (Cav2+) played an important role in the neuronal differentiation of NSCs under ES. Our findings demonstrated that ES combined with three-dimensional conductive scaffolds would be a promising strategy to regulate the neuronal differentiation of NSCs for neural regeneration.

Selenite Ameliorates Cadmium-induced Cytotoxicity Through Downregulation of ROS Levels and Upregulation of Selenoprotein Thioredoxin Reductase 1 in SH-SY5Y Cells.

Cadmium (Cd) as a ubiquitous toxic heavy metal in the environment, causes severe hazards to human health, such as cellular stress and organ injury. Selenium (Se) was reported to reduce Cd toxicity and the mechanisms have been intensively studied so far. However, it is not yet crystal clear whether the protective effect of Se against Cd-induced cytotoxicity is related to selenoproteins in nerve cells or not. In this study, we found that Cd inhibited selenoprotein thioredoxin reductase 1 (TrxR1; TXNRD1) and decreased the expression level of TrxR1, resulting in cellular oxidative stress, and Se supplements ameliorated Cd-induced cytotoxicity in SH-SY5Y cells. Mechanistically, the detoxification of Se against Cd is attributed to the increase of the cellular TrxR activity and upregulated TrxR1 protein level, culminating in strengthened antioxidant capacity. Results showed that Se supplements attenuated the ROS production and apoptosis in SH-SY5Y cells, and significantly mitigated Cd-induced SH-SY5Y cell death. This study may be a valuable reference for shedding light on the mechanism of Cd-induced cytotoxicity and the role of TrxR1 in Se-mitigated cytotoxicity of Cd in neuroblast cells, which may be helpful for understanding the therapeutic potential of Cd and Se in treating or preventing neurodegenerative diseases, like Alzheimer's disease (AD) and Parkinson's disease (PD).

Synthesis and characterization of protocatechuic acid grafted carboxymethyl chitosan with oxidized sodium alginate hydrogel through the Schiff's base reaction.

Excessive accumulation of free radicals is closely related to the occurrence and development of various neurodegenerative diseases. In this study, a novel protocatechuic acid grafted carboxymethyl chitosan with oxidized sodium alginate (PCA-g-CMCS/OSA) hydrogel was developed to maintain the oxidation-antioxidation balance activities. By optimizing the pH-soluble range (pH > 6.4) of CMCS, PCA was grafted onto CMCS skeleton via EDC/NHS, and then conjugated with aldehyde group of OSA to form Schiff's base hydrogel at physiological temperature. The gelation time can be adjusted rapidly within 1-3 min by controlling the content of OSA. The shaped hydrogel exhibited porous network structure with high porosity (>90 %), swelling ratio (2000-3000 %) and rheological property, which is beneficial to cell growth and proliferation. The conjugates preserved excellent DPPH and ABTS radicals scavenging abilities and adequate biodegradability within 5 weeks. Moreover, with the release of PCA monomer due to degradation of the PCA-g-CMCS/OSA, the hydrogel also exhibited excellent biocompatibility and protective effect on H2O2-induced oxidative damage in PC12 cells. These results suggested that the PCA-g-CMCS/OSA hydrogel would appear to be a more attractive candidate for potential biomedical applications such as antioxidant drug release and tissue engineering implant material.

NIHSS-the Alberta Stroke Program Early CT Score mismatch in guiding thrombolysis in patients with acute ischemic stroke.

OBJECTIVE: This study investigates the mismatch between the National Institutes of Health Stroke Scale (NIHSS) score and the computed tomography (CT) findings measured by the Alberta Stroke Program Early CT Score (ASPECTS) for predicting the functional outcome and safety of intravenous thrombolysis (IVT) treatment in patients with acute ischemic stroke (AIS). METHODS: This prospective observational study includes patients with AIS who underwent CT imaging within 4.5 h of the onset of symptoms. Patients were divided into the NIHSS-ASPECTS mismatch (NAM)-positive and NAM-negative groups (group P and N, respectively). The clinical outcome was assessed using the Modified Rankin Scale (mRS). Safety outcomes included progression, symptomatic intracerebral hemorrhage (sICH), intracerebral hemorrhage (ICH), adverse events, clinical adverse events, and mortality. RESULTS: A total of 208 patients were enrolled in the study. In group P, IVT treatment was associated with a good functional outcome at 3 months (p = 0.005) and 1 year (p = 0.001). A higher percentage of patients with favorable mRS (0-2) (p = 0.01) and excellent mRS (0-1) (p = 0.011) functional outcomes was obtained at 1 year in group P with IVT treatment. Group N did not benefit from the same treatment (p = 0.352 and p = 0.480 at 3 months and 1 year, respectively). There were no statistically significant differences in sICH, ICH, mortality rates, or other risks between the IVT and conventional treatment groups. CONCLUSION: IVT treatment is associated with a good functional outcome in patients with NAM, without increasing the risks of sICH, ICH, mortality, or other negative outcomes. NAM promises to be an easily obtained indicator for guiding the treatment decisions of AIS.

Overexpression of human MRP1 in neurons causes resistance to antiepileptic drugs in Drosophila seizure mutants.

Multidrug resistance-associated protein 1 (MRP1), an efflux multidrug transporter, was shown to be elevated in both glia and neurons in seizure focus in refractory epilepsy patients. Up-regulation of MRP1 and other multidrug transporters in perivascular astrocytes was suggested to cause resistance to antiepileptic drugs (AEDs) by reducing the concentration of AEDs at the epileptogenic areas. However, it is not known whether the up-regulation of MRP1 in neurons can cause resistance to AEDs, such as sodium phenytoin (PHT) and valproic acid (VPA). PHT inhibits voltage-gated sodium channel (VGSC) by occluding it, but whether PHT enters the channel through its inner or outer pore is not known. The authors overexpressed human MRP1 protein only in neurons in a Drosophila genetic seizure model, bang senseless (bss) mutants. The authors found that overexpression of MRP1 blocked the attenuation of the seizure behavior of bss mutants by acute and chronic application of PHT, and by chronic application of VPA. Conversely, overexpression of MRP1 in neurons increased the tolerance of bss flies to high-dosage PHT and VPA. Thus, up-regulation of MRP1 expression only in neurons causes resistance to AED in seizure flies. Moreover, the current data suggest that PHT enters VGSC through its inner pore.

Isomer-specific effects of conjugated linoleic acid on proliferative activity of cultured neural progenitor cells.

Conjugated linoleic acid (CLA) has been shown to have a variety of biological activities. However, the effects of CLA on the proliferation of neural progenitor cells (NPCs) are not clear. The objective of this study was to determine the effects of cis-9 trans-11 CLA and trans-10 cis-12 CLA, the predominant individual isomers, on the proliferative activity of NPCs in vitro. Cell counts showed that treatment of NPCs with cis-9 trans-11 CLA increased the cell number in a dose- and time- dependent manner while significant inhibition effect of trans-10 cis-12 CLA was observed. Western blot analysis revealed the elevated expression of cyclin D1 induced by cis-9 trans-11 CLA treatment and the decreased expression of cyclin D1 by trans-10 cis-12 CLA treatment in NPCs. Cyclin D1-siRNA transfection significantly inhibited the promotion of cell proliferation by cis-9 trans-11 CLA. In addition, trans-10 cis-12 CLA inhibited the phosphorylation of protein kinase B (PKB/Akt), while cis-9 trans-11 CLA had no effect on phospho Akt levels. Furthermore, immunofluorescence assay showed that after CLA treatment, the cells retained their functional characteristics of neural progenitors. These results indicated that cis-9 trans-11 CLA can effectively enhance the proliferation of hADSCs. The effect of cis-9 trans-11 CLA may be associated with the up-regulation of cyclin D1 expression.

Anti-ageing effects of protocatechuic acid from Alpinia on spleen and liver antioxidative system of senescent mice.

The effects of Alpinia protocatechuic acid (PCA) on spleen and liver antioxidant system in aged rats have been studied. Alpinia PCA, a phenolic compound, was first isolated from the dried fruits of Alpinia Oxyphylla Miq. in our laboratory. Young and aged rats were injected intraperitoneally with Alpinia PCA at single doses of 5 mg kg(-1) (low dose) or 10 mg kg(-1) (high dose) per day for 7 days. The activities of endogenous antioxidants and the content of lipid peroxide in spleen and liver were assayed. Compared with young group, aged rats had significantly lower splenic weights, lower activities of glutathione peroxidase (GSH-PX) and catalase (CAT), higher level of malondialdehyde (MDA) in spleen and liver. The results proved that Alpinia PCA significantly elevated the splenic weights, increased the activities of GSH-PX and CAT and decreased the MDA level of aged rats. All these suggested that Alpinia PCA was a potential anti-ageing agent, and its effects on spleen and liver were achieved at least partly by promoting endogenous antioxidant enzymatic activities and normalizing age-associated alterations. It may be therapeutically useful to minimize age-associated disorders where oxidative damage is the major cause.

Preparation, characterization and antioxidant activity of protocatechuic acid grafted carboxymethyl chitosan and its hydrogel.

In this study, protocatechuic acid (PCA) was grafted onto carboxymethyl chitosan (CMCS) via EDC/NHS to improve the antioxidant effect. The grafting ratio of PCA-g-CMCS conjugates could be controlled by adjusting the pH value and feed ratio of raw materials. The conjugates exhibited similar pH sensitivity to CMCS and showed dramatic enhancements of DPPH and ABTS radicals scavenging activities, total antioxidant capacity, reducing power, and Fe2+-chelating activity. Three-dimensional porous PCA-g-CMCS hydrogel was prepared by lyophilization and secondary cross-linking. The shaped hydrogel preserved its antioxidant activity, and the sustained release of PCA-containing degraded fragment from biodegradable hydrogel could be achieved with the aid of lysozyme in vitro (15 days). PCA-g-CMCS hydrogel also showed excellent biocompatibility and protective effect on H2O2-induced oxidative damage in SH-SY5Y cells. These results suggested that PCA-g-CMCS conjugates and its hydrogel would appear to be a promising oxidation-resistant material for applications such as drug release and tissue engineering.

Chlorophyllin Inhibits Mammalian Thioredoxin Reductase 1 and Triggers Cancer Cell Death.

Food colorants are widely used by humans in food production and preparation; however, their potential toxicity requires an in-depth analysis. In this study, five out of 15 commercial food colorants, namely, lutein, betanin, caramel, crocin and chlorophyll, significantly inhibited wild type selenoprotein thioredoxin reductase 1 (TrxR1, TXNRD1) in vitro. The hyperactive Sec498 residue of TrxR1 was targeted by those five colorants, which was confirmed by the site-directed mutagenesis of TrxR1. Furthermore, two colorants, chlorophyll and betanin, triggered the oligomerization of TrxR1. A chlorophyll-derived compound, chlorophyllin, irreversibly inhibited the 5,5'-dithiobis-2-nitrobenzoic acid (DTNB) reducing activity of TrxR1 with Kinact = 6.96 × 10-3 ± 0.49 × 10-3 µM-1 min-1. Moreover, chlorophyllin reduced the cellular TrxR activity, leading to reactive oxygen species (ROS) accumulation and, subsequently, promoting cancer cell death. In conclusion, this study might contribute to understand the food safety of commercial colorants and provide chemotherapeutic compounds by targeting TrxR1.

Effect of platelet count on long-term prognosis of cerebral infarction.

OBJECTIVE: This study aimed to analyze the correlation between platelet (PLT) count and the modified Rankin scale (mRS) in patients with cerebral infarction (CI) at the later stage of rehabilitation, which can be used to guide the secondary prevention strategy of CI. METHODS: A total of 180 CI patients were divided into three groups according to PLT count: low PLT group (<125×109/L), medium PLT group (126- 225×109/L) and high PLT group (>226×109/L). The mRS was evaluated after three months and one year, respectively, and the difference in long-term prognosis between groups was analyzed. The mRS is an ordered scale coded from 0 (no symptoms at all) through 5 (severe disability) 6 (death). RESULTS: Finally, a total of 99 patients had complete data. The results of the multiple comparisons among the three groups were as follows: the analysis of variance of the mRS at three months after onset yielded F = 6.714 and P = 0.002, and the difference was statistically significant. The mRS was lowest in the medium PLT group (2.09±1.465), and neurological function recovery was the best. After one year, the mRS for the medium PLT group was the lowest (1.49±1.523), with F = 6.860 and P = 0.002. The repeated measures analysis of variance revealed that the effect of continuous rehabilitation was significant in the interval from three months to one year after onset (F = 35.528, P < 0.001). This was very significant, especially for patients taking aspirin (F = 50.908, P < 0.001). However, for patients who did not take aspirin, the effect of continuous rehabilitation was not obvious during the nine months, and the difference between the results of two mRS measurements was not statistically significant (F = 1.089, P = 0.308). CONCLUSIONS: Patients with a PLT count of 126- 225×109/L had the lowest mRS between three months and one year after onset, but had the best recovery of nerve function. Patients who persisted in taking aspirin continued to significantly recover during the 9-month period, from three months to one year after onset. Aspirin is not only a secondary preventive drug, but also an important drug to promote the rehabilitation of CI patients.

Effect of protocatechuic acid from Alpinia oxyphylla on proliferation of human adipose tissue-derived stromal cells in vitro.

The effect of protocatechuic acid (PCA) from Alpinia oxyphylla and catapol from Rehmannia on the proliferation capacity of human adipose tissue-derived stromal cells (hADSCs) was investigated in vitro. Cell counts showed that treatment of hADSCs with PCA for 48 h increased the cell number in a dose-dependent manner, while no obvious effect of catapol on the proliferation of hADSCs was observed. In addition, the cell number of hADSCs treated by 1.5 mM PCA increased in a time-dependent manner. The flow cytometric analysis of DNA content demonstrated the cell cycle progress from the G0/G1 phase to the S phase. Western blot analysis revealed the elevated expression of cyclin D1 in hADSCs induced by PCA treatment. Cyclin D1-siRNA transfection significantly inhibit the promotion of cell proliferation by PCA. Furthermore, the flow cytometric analysis of the cell surface antigens and the multidifferential potential tests of PCA-treated hADSCs showed that the cells retained their functional characteristics of multipotential mesenchymal progenitors. It is concluded that PCA can effectively up-regulate the proliferation of hADSCs.

Protocatechuic acid promotes cell proliferation and reduces basal apoptosis in cultured neural stem cells.

Protocatechuic acid (PCA), a phenolic compound isolated from the kernels of Alpinia oxyphylla, showed anti-oxidant neuroprotective property in our previous study. However, it is still unknown whether PCA have effects on the cultured neural stem cells (NSCs). In this study, we investigated the roles of PCA in the survival and apoptosis of rat NSCs under normal conditions. NSCs obtained from 13.5-day-old rat embryos were propagated as neurospheres and cultured under normal conditions with or without PCA for 4 and 7 days. The cell viability was determined by the cell counting kit-8 (CCK-8) test, while cell proliferation was assayed by bromodeoxyuridine (BrdU) labeling. PCA increased the cellular viability of NSCs and stimulated cell proliferation in a dose- and time-dependent manner. Apoptotic cells were detected after 4 days by observing the nuclear morphological changes and flow cytometric analysis. Compared with the control on both culture days, treatment with PCA effectively reduced the levels of apoptosis of NSCs. At the same time, the reactive oxygen species (ROS) level in NSCs was depressed. In addition, PCA also significantly decreased the activity of elevated caspase-3, indicating that PCA may inhibit apoptosis of NSCs via suppression of the caspase cascade. These results suggest that PCA may be a potential growth inducer and apoptosis inhibitor for NSCs.

Mechanism Study of Bacteria Killed on Nanostructures.

It is important to study the bactericidal mechanism with nanostructures for the design and preparation of high-efficiency sterilization materials. In this paper, the interfacial energy gradient between cells and nanopillars is proposed to be the driving force to promote cells to migrate into nanostructures and get killed. The expressions of interfacial energy and its gradient were first established, then the deformation of cells pressured by nanostructures was calculated. The results show that the interfacial energy gradient or the pressure on cells is influenced by nanopillar parameters substantially. The smaller the nanopillar diameter and the larger the pitch, the greater the pressure on cells. Only high enough nanocolumns can ensure sufficient cell creep deformation and become punctured. Furthermore, a cell volume and its adhesion morphology also influence the interaction between cells and nanostructures. The smaller the cell volume, the greater the pressure on it. And the larger the contact angle of adhered cells, the greater the pressure on the cells by nanopillars. Besides, the wettability of substrate material also influences the interaction between cells and nanopillars. It can be concluded that the model is reasonable and reliable since its calculation results are in good accordance with the experimental measurements.

Higher Platelet-to-Lymphocyte Ratio Is Associated With Worse Outcomes After Intravenous Thrombolysis in Acute Ischaemic Stroke.

Objective: The platelet-to-lymphocyte ratio (PLR) is a new marker of atherosclerotic inflammation and has been identified as a predictive factor in cardiovascular diseases, but its significance in patients with acute ischaemic stroke (AIS) who have undergone intravenous thrombolysis (IVT) is still unknown. Methods: Consecutive patients who were treated with IVT using recombinant tissue plasminogen activator (rtPA) for AIS were included from May 2012 to August 2018. The PLR was calculated according to platelet and lymphocyte counts within 24 h after thrombolysis therapy. Functional outcomes were assessed by the modified Rankin Scale (mRS) at 3 months after thrombolysis. Stroke severity was assessed by National Institutes of Health Stroke Scale (NIHSS) scores. The primary endpoint was an unfavorable outcome (mRS > 2), and the secondary endpoint was death at 3 months. Results: A total of 286 patients were included in the study. The median age was 69.5 (59.0-80.0) years, and 59.1% of patients were men. A total of 120 (42.0%) patients had an unfavorable outcome, and 38 (13.2%) died. Patients with an unfavorable outcome had significantly higher PLR values compared with those with a favorable outcome [172.5 (105.3-239.0) vs. 139 (97.0-194.5), P = 0.008], and the PLR values of the patients who died at 3 months were higher than those of the surviving patients [189.5 (127.5-289.0) vs. 142.0 (98.0-215.5), P = 0.006]. After adjustment for other variables, the PLR was independently associated with the two endpoints: unfavorable outcome (OR 2.220, 95% CI 1.245-3.957, P = 0.007) and death (OR 2.825, 95% CI 1.050-7.601, P = 0.040) at 3 months after thrombolysis. In addition, PLR was correlated with the NIHSS score (R = 0.230, P < 0.001). Conclusions: Higher PLR levels were independently associated with an unfavorable outcome and death at 3 months in AIS patients treated with IVT.

Protocatechuic acid from Alpinia oxyphylla promotes migration of human adipose tissue-derived stromal cells in vitro.

Human adipose tissue-derived stromal cells (hADSCs) demonstrate promising potential in various clinical applications, including the transplantation to regenerate injured or degenerative tissues. The migration of engrafted hADSCs to the correct site of injure is essential for the curative effect of stem cell therapy. We found that protocatechuic acid (PCA) from Alpinia (A.) oxyphylla could promote the migration capacity of hADSCs through transwell coated with gelatin in vitro. PCA enhanced the cell migration rate in a dose-dependent and time-dependent manner. Meanwhile, RT-PCR and quantitative RT-PCR analysis revealed the elevation of membrane-type matrix metalloproteinase-1 (MT1-MMP) mRNA expression in 1.5 mM PCA-treated hADSCs. In the supernatants of these cells, the active matrix metalloproteinase-2 (MMP-2) increased compared with control cells with zymography. Moreover, the promotion of cell migration by PCA could be effectively and obviously inhibited by anti-MT1-MMP or anti-MMP-2 antibodies. Furthermore, flow cytometric analysis of the cell surface antigens, osteogenic induction, adipogenic induction and cardiomyocyte-like cell induction demonstrated that hADSCs retained their functional characteristics of multipotential mesenchymal progenitors after PCA treatment. These results suggest that PCA from A. oxyphylla promote the migration of hADSCs in vitro, which is partially due to the increased expression of MT1-MMP and the promotion of MMP-2 zymogen activation.

3D culture of neural stem cells within conductive PEDOT layer-assembled chitosan/gelatin scaffolds for neural tissue engineering.

Neural stem cells (NSCs), as a self-renewing and multipotent cell population, have been widely studied for never regeneration. Engineering scaffold is one of the important factors to regulate NSCs proliferation and differentiation towards the formation of the desired cells and tissues. Because neural cells are electro-active ones, a conductive scaffold is required to provide three-dimensional cell growth microenvironments and appropriate synergistic cell guidance cues. In this study, a poly (3,4­ethylenedioxythiophene)/chitosan/gelatin (PEDOT/Cs/Gel) scaffold was prepared via in situ interfacial polymerization, with a nanostructured layer of PEDOT assembling on the channel surface of porous Cs/Gel scaffold. This electrically conductive, three-dimensional, porous and biodegradable PEDOT/Cs/Gel scaffold was used as a novel scaffold for NSCs three-dimension (3D) culture in vitro. It was found that the layer of PEDOT on the channel surface of Cs/Gel scaffolds could greatly promote NSCs adhesion and proliferation. Additionally, under the differentiation condition, the protein and gene analysis suggested that PEDOT/Cs/Gel scaffolds could significantly enhance the NSCs differentiation towards neurons and astrocytes with the up-regulation of ß tubulin-III and GFAP expression. In conclusion, these results demonstrated that the PEDOT/Cs/Gel scaffolds as an electrically conductive scaffold could not only promote NSCs adhesion and proliferation but also enhance NSCs differentiation into neurons and astrocytes with higher protein and gene expression. PEDOT-assembled Cs/Gel scaffold will be a promising conductive substrate for NSCs research and neural tissue engineering.