Amelioration effects of the soybean lecithin-gallic acid complex on iron-overload-induced oxidative stress and liver damage in C57BL/6J mice.

CONTEXT: Gallic acid (GA) and lecithin showed important roles in antioxidant and drug delivery, respectively. A complex synthesized from GA and soybean lecithin (SL-GAC), significantly improved bioavailability of GA and pharmacological activities. However, the antioxidant activity of SL-GAC and its effect on iron-overload-induced liver injury remains unexplored. OBJECTIVE: This study investigates the antioxidant properties of SL-GAC in vitro and in mice, and its remediating effects against liver injury by iron-overloaded. MATERIALS AND METHODS: In vitro, free radical scavenging activity, lipid peroxidation inhibition, and ferric reducing power of SL-GAC were measured by absorbance photometry. In vivo, C57BL/6J mice were randomized into 4 groups: control, iron-overloaded, iron-overloaded + deferoxamine, and iron-overloaded + SL-GAC. Treatments with deferoxamine (150 mg/kg/intraperitioneally) and SL-GAC (200 mg/kg/orally) were given to the desired groups for 12 weeks, daily. Iron levels, oxidative stress, and biochemical parameters were determined by histopathological examination and molecular biological techniques. RESULTS: In vitro, SL-GAC showed DPPH and ABTS free radicals scavenging activity with IC50 values equal to 24.92 and 128.36 µg/mL, respectively. In C57BL/6J mice, SL-GAC significantly reduced the levels of serum iron (22.82%), liver iron (50.29%), aspartate transaminase (25.97%), alanine transaminase (38.07%), gamma glutamyl transferase (42.11%), malondialdehyde (19.82%), total cholesterol (45.96%), triglyceride (34.90%), ferritin light chain (18.51%) and transferrin receptor (27.39%), while up-regulated the levels of superoxide dismutase (24.69%), and glutathione (11.91%). CONCLUSIONS: These findings encourage the use of SL-GAC to treat liver injury induced by iron-overloaded. Further in vivo and in vitro studies are needed to validate its potential in clinical medicine.

2†µm sub-GHz harmonic mode-locked soliton generation based on a Bi2S3 saturable absorber.

Saturable absorber (SA) based harmonic mode-locking (HML) techniques at 2 µm waveband are much less reported than those at 1.5 µm waveband, the maximum repetition rate of the harmonic pulse generated by such techniques at 2 µm waveband is also much lower than those generated at 1.5 µm waveband. In this paper, the 39th harmonic with the repetition rate of 908.6 MHz is realized in a Bi2S3-based thulium-doped fiber laser. The fundamental mode-locked pulse has a central wavelength of 1954.2 nm and a 3-dB bandwidth of 5.1 nm. The repetition rate is 23.27 MHz and the pulse width is 902 fs. The characteristics of the material and harmonic mode-locked pulse are investigated. To the best of our knowledge, this is the highest and the closest resonance frequency to GHz among the reported SA-based harmonic mode-locked fiber lasers operating at 2 µm waveband.

Simulation and analysis of supercontinuum generation in the waveband up to 25 µm.

The ultra-wideband supercontinuum generation (SCG) in a Te-based chalcogenide (ChG) photonic crystal fiber (PCF) is simulated in the mid-infrared (MIR) waveband. The PCF core and cladding materials are Ge20As20Se15Te45 and Ge20As20Se17Te43, respectively. The supercontinuum (SC) broadening affected by the core diameter and fiber absorption is considered. The selected PCFs at different pumping wavelengths can demonstrate the generation of ultra-wideband MIR supercontinuum according to the simulated results. We consider SC broadening with and without fiber absorption. A SC range from 3 to 25 µm is demonstrated by simulation in a PCF with a core diameter of 8 µm and a pump wavelength of 6 µm considering the fiber absorption. With the increase of the peak power and the pulse width and the decrease of the core diameter, the degree of coherence gradually degraded. To the best of our knowledge, this is the first demonstration of the possibility of SCG up to the waveband of 25 µm in fiber. Our results highlight the potential of a novel Te-based chalcogenide multi-material PCF for SCG. We also provide a way to generate the SCs to longer wavebands than 20 µm in fiber, especially up to the far-infrared waveband.

The difference of gut microbiome in different biliary diseases in infant before operation and the changes after operation.

BACKGROUND: Evidence supports an association between cholestatic liver disease and changes in microbiome composition. Nevertheless, the identification of this special type of biliary atresia from non-biliary atresia cholestasis is still a major clinical difficulty. The purpose of this study is to compare the differences in the composition of gut microbiome between infants with biliary atresia and infant with non-biliary atrestic cholestasis, to find new ways to identify and diagnose these two diseases early, to understand the influence of the presence or absence of bile on the composition of the gut microbiome in infants with cholestasis. METHODS: Using 16S rDNA gene sequencing technology to analyze the intestinal flora of the participants. RESULTS: In terms of diversity, there is an obvious structural separation in the intestinal microbiota of the BA group and the CD group, and this structural separation also exists in the comparison between the two groups before surgery. Taxonomic analysis demonstrated that the two groups showed an increase in Proteobacteria and Firmicutes before surgery, and the relative abundance of potential pathogens such as Shigella, Streptococcus, Klebsiella, etc. increased, potential probiotics such as Bifidobacteria and Lactobacillus decreased, but the relative abundance of each genus was different between groups. It was found that Enterococcus, Ralstonia, Nitriliruptoraceae, etc. were differentially enriched in the BA group, the CD group are mainly enriched in Veillonella, Clostridium_sensu_stricto_1 and Lactobacillus. Functional analysis of the groups showed that the BA group mainly focused on the processes of energy release processes, and the CD group mainly focused on the biosynthesis of amino-acids to consume energy. CONCLUSIONS: The composition of intestinal flora is different between biliary atresia and non-biliary atretic cholestasis. Enterococcus, Ralstonia, etc. may become biomarkers for the identification and diagnosis of both.

Transcriptome analysis reveals dynamic changes in coxsackievirus A16 infected HEK 293T cells.

BACKGROUND: Coxsackievirus A16 (CVA16) and enterovirus 71 (EV71) are two of the major causes of hand, foot and mouth disease (HFMD) world-wide. Although many studies have focused on infection and pathogenic mechanisms, the transcriptome profile of the host cell upon CVA16 infection is still largely unknown. RESULTS: In this study, we compared the mRNA and miRNA expression profiles of human embryonic kidney 293T cells infected and non-infected with CVA16. We highlighted that the transcription of SCARB2, a cellular receptor for both CVA16 and EV71, was up-regulated by nearly 10-fold in infected cells compared to non-infected cells. The up-regulation of SCARB2 transcription induced by CVA16 may increase the possibility of subsequent infection of CVA16/EV71, resulting in the co-infection with two viruses in a single cell. This explanation would partly account for the co-circulation and genetic recombination of a great number of EV71 and CVA16 viruses. Based on correlation analysis of miRNAs and genes, we speculated that the high expression of SCARB2 is modulated by down-regulation of miRNA has-miR-3605-5p. At the same time, we found that differentially expressed miRNA target genes were mainly reflected in the extracellular membrane (ECM)-receptor interaction and circadian rhythm pathways, which may be related to clinical symptoms of patients infected with CVA16, such as aphthous ulcers, cough, myocarditis, somnolence and potentially meningoencephalitis. The miRNAs hsa-miR-149-3p and hsa-miR-5001-5p may result in up-regulation of genes in these morbigenous pathways related to CVA16 and further cause clinical symptoms. CONCLUSIONS: The present study elucidated the changes in 293T cells upon CVA16 infection at transcriptome level, containing highly up-regulated SCARB2 and genes in ECM-receptor interaction and circadian rhythm pathways, and key miRNAs in gene expression regulation. These results provided novel insight into the pathogenesis of HFMD induced by CVA16 infection.

[NK-92 cells modified with chimeric antigen receptor targeting PD-L1 inhibits the proliferation of A549 lung cancer cells].

Objective To investigate the anti-tumor activity of PD-L1-redirected chimeric antigen receptor modified NK-92 cells in lung cancer. Methods NK-92 cells modified by chimeric antigen receptor (pCAR-92) was obtained by lentivirus transfection. Tumor cells overexpressing PD-L1 were induced by IFN-γ. Lactic dehydrogenase (LDH) level was used cells to identify the cytotoxicity of pCAR-92 cells to target cells. The activation markers of CD107a and IFN-γ in pCAR-92 cells were detected by flow cytometry, and the anti-tumor activity of pCAR-92 cells in vivo was verified by xenograft model. Results Flow cytometry showed that the positive ratio of pCAR-92 cells ranged from 70% to 80%. The LDH detection showed that pCAR-92 cells could significantly lyse tumor cells induced by IFN-γ compared to control group. Flow cytometry for the expression of CD107a and IFN-γ showed that pCAR-92 cells could be significantly activated after co-incubation with tumor cells induced by IFN-γ. The tumor inhibitory effect of pCAR-92 cells was stronger than that of control group. In addition, after pCAR-92 cells treatment, the expression of PD-L1 in tumor decreased and the number of tumor infiltrating NK cells increased. Conclusion NK-92 cells modified with chimeric antigen receptors targeting PD-L1 have evident anti-tumor effects.

The effect of reverse intergenerational support on the happiness of Chinese older adults: A moderating effect analysis based on role conflict.

Intergenerational support is bidirectional, and reverse intergenerational support refers to parents providing financial support, time support, and spiritual support to their offspring. The emergence of reverse intergenerational support has created role conflicts among different groups of older adults. Based on survey data from 3,170 elderly people in eight sample provinces in China, this paper empirically investigates the relationship between reverse intergenerational support and the happiness of the elderly in contemporary China and the moderating effect of role conflict in it, using an ordered logit model. It was found that, first, reverse economic support reduces the happiness of the elderly, and reverse time support and reverse spiritual support can significantly enhance the happiness of the elderly. Second, in the presence of role conflict, the effect of reverse time support and reverse spiritual support on the enhancement of older adults' happiness was suppressed; in the presence of role enhancement, the effect of reverse economic support on the reduction of older adults' happiness was mitigated. The above findings provide new empirical evidence for understanding the relationship between reverse intergenerational support and the happiness of the elderly, which is prevalent in contemporary China, and offer new insights for enhancing happiness.

Association between the Phytochemical Index and Overweight/Obesity: A Meta-Analysis.

Some studies suggest that a higher phytochemical index (PI) is associated with a lower risk of overweight/obesity. This meta-analysis is performed to summarize published studies on the relationship of PI and the risk of overweight/obesity. We searched on PubMed, Cochrane Library and Web of Science from the inception dates to February 2022. The random-effect model was used based on heterogeneity. Meta-regression was used to explore potential sources of between-study heterogeneity. Publication bias was evaluated using Begg's and Egger's tests. The dose-response relationship was assessed using a restricted cubic spline model. Nine studies were included in the meta-analysis, with a total of 100,753 participants. The meta-analysis showed that the phytochemical index was associated with a decreased risk of overweight/obesity. The pooled OR (95% CI) was 0.81 (0.74-0.90). The findings from dose-response analysis showed a nonlinear association between the phytochemical index and the risk of overweight/obesity. The results of the meta-regression showed that gender and area were significant covariates influencing the heterogeneity between studies. There was no publication bias in the meta-analysis of this study. In conclusion, although this meta-analysis indicates that a high phytochemical index is associated with a reduced risk of overweight/obesity, all the studies included in this meta-analysis were cross-sectional studies with high heterogeneity. As such, more data from randomized controlled trials are required to confirm the efficacy of PI in evaluating the risk of overweight/obesity.

Antiviral Efficacy of Flavonoids against Enterovirus 71 Infection in Vitro and in Newborn Mice.

Enterovirus 71 (EV71) infection is known to cause hand, foot, and mouth disease (HFMD), which is associated with neurological complications; however, there is currently no effective treatment for this infection. Flavonoids are a large group of naturally occurring compounds with multiple bioactivities, and the inhibitory effects of several flavonoids against EV71 have been studied in cell cultures; however, to date, there are no reported data on their effects in animal models. In this study, we confirmed the in vitro activities of eight flavonoids against EV71 infection, based on the inhibition of cytopathic effects. Moreover, these flavonoids were found to reduce viral genomic RNA replication and protein synthesis. We further demonstrated the protective efficacy of these flavonoids in newborn mice challenged with a lethal dose of EV71. Apigenin, luteolin, kaempferol, formononetin, and penduletin conferred survival protection of 88.89%, 91.67%, 88.89%, 75%, and 66.67%, respectively, from the lethal EV71 challenge. In addition, isorhamnetin provided the highest mice survival protection of 100% at a dose of 10 mg/kg. This study, to the best of our knowledge, is the first to evaluate the in vivo anti-EV7l activities of multiple flavonoids, and we accordingly identified flavonoids as potential leading compounds for anti-EV71 drug development.

Exosomes Released from Rabies Virus-Infected Cells May be Involved in the Infection Process.

Exosomes are cell-derived vesicles that are secreted by many eukaryotic cells. It has recently attracted attention as vehicles of intercellular communication. Virus-infected cells release exosomes, which contain viral proteins, RNA, and pathogenic molecules. However, the role of exosomes in virus infection process remains unclear and needs to be further investigated. In this study, we aimed to evaluate the effects of exosomes on rabies virus infection. OptiPrep™ density gradient centrifugation was used to isolate exosomes from rabies virus-infected cell culture supernatants. A rabies virus G protein enzyme-linked immunosorbent assay and acetylcholinesterase activity assays were performed to verify the centrifugation fractions. Exosomes were then characterized using transmission electron microscopy and Western blotting. Our results showed that rabies virus infection increased the release of exosomes. Treatment with GW4869 and si-Rab27a, two exosomal secretion inhibitors, inhibited exosome release. Furthermore, the inhibitors reduced the levels of extracellular and intracellular viral RNA. These data indicated that exosomes may participate in the viral infection process. Moreover, our results establish a basis for future research into the roles of exosomes in rabies virus infection and as potential targets for developing new antiviral strategies.

Antiviral Effects of ABMA against Herpes Simplex Virus Type 2 In Vitro and In Vivo.

Herpes simplex virus type 2 (HSV-2) is the causative pathogen of genital herpes and is closely associated with the occurrence of cervical cancer and human immunodeficiency virus (HIV) infection. The absence of an effective vaccine and the emergence of drug resistance to commonly used nucleoside analogs emphasize the urgent need for alternative antivirals against HSV-2. Recently, ABMA [1-adamantyl (5-bromo-2-methoxybenzyl) amine] has been demonstrated to be an inhibitor of several pathogens exploiting host-vesicle transport, which also participates in the HSV-2 lifecycle. Here, we showed that ABMA inhibited HSV-2-induced cytopathic effects and plaque formation with 50% effective concentrations of 1.66 and 1.08 µM, respectively. We also preliminarily demonstrated in a time of compound addition assay that ABMA exerted a dual antiviral mechanism by impairing virus entry, as well as the late stages of the HSV-2 lifecycle. Furthermore, in vivo studies showed that ABMA protected BALB/c mice from intravaginal HSV-2 challenge with an improved survival rate of 50% at 5 mg/kg (8.33% for the untreated virus infected control). Consequently, our study has identified ABMA as an effective inhibitor of HSV-2, both in vitro and in vivo, for the first time and presents an alternative to nucleoside analogs for HSV-2 infection treatment.

Synonymous codon usage analysis of hand, foot and mouth disease viruses: A comparative study on coxsackievirus A6, A10, A16, and enterovirus 71 from 2008 to 2015.

Enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) have been considered major pathogens of hand, foot and mouth disease (HFMD) throughout the world for decades. In recent years, coxsackievirus A6 (CVA6) and coxsackievirus A10 (CVA10) have raised attention as two other serious pathogens of HFMD. The present study focused on the synonymous codon usage of four viruses isolated from 2008 to 2015, with particular attention on P1 (encoding capsid proteins) and P2-P3 regions (both encoding non-structural proteins) in the genomic RNA. Relative synonymous codon usage, effective number of codons, neutrality and correspondence were analyzed. The results indicated that these viruses prefer A/T at the third position in codons rather than G/C. The most frequent codons of 4 essential and 2 semi-essential amino acids, as well as a key amino acid of metabolic junctions (Glu) used in the four viruses are also the most frequently used in humans. Effective number of codons (ENC) values indicated weak codon usage bias in all the viruses. Relatively, the force of mutation pressure in the P1 region was found to be stronger than that in the P2-P3 region, and this force in the P1 region of CVA6 and EV71 was stronger than that of CVA10 and A16. The neutrality analysis results implied that mutation pressure plays a minor role in shaping codon bias of these viruses. Correspondence analysis indicated that the codon usage of EV71 strains varied much more than that of other viruses. In conclusion, the present study provides novel and comparative insight into the evolution of HFMD pathogens at the codon level.

Antiviral effects of Retro-2cycl and Retro-2.1 against Enterovirus 71 in vitro and in vivo.

Enterovirus 71 (EV71) is one of the causative pathogens of hand, foot and mouth disease (HFMD), especially the form associated with fatal neurological disorders. Sustained outbreaks of EV71 infections remain a serious health threat worldwide. However, no antiviral agent against EV71 for clinical therapy has been approved. Retro-2cycl and Retro-2.1 are inhibitors of several pathogens specifically targeting the intracellular vesicle transport, which also participates in the EV71 lifecycle processes including progeny virus release. Here, we reported that Retro-2cycl and Retro-2.1, respectively, could inhibit EV71 infection with 50% effective concentrations of 12.56 µM and 0.05 µM in a cytopathic effect inhibition assay and showed relatively low cytotoxicity with 50% cytotoxicity concentrations of more than 500 µM and 267.80 µM. Preliminary mechanism studies revealed that Retro-2cycl and Retro-2.1 did not inhibit EV71 protein synthesis or RNA replication but could block progeny EV71 release specifically. Furthermore, administration of Retro-2cycl at the dose of 10 mg/kg significantly protected 90% of newborn mice from lethal EV71 challenge. Consequently, our results for the first time identified Retro-2cycl and Retro-2.1 as effective inhibitors of EV71 as well as lead compounds, which would contribute to anti-EV71 drug development. We also identified progeny virus release and the intracellular vesicle transport as antiviral targets for EV71.

Preparation of a mixed-matrix hydrogel of vorinostat for topical administration on the rats as experimental model.

Oral vorinostat has the remarkable curative effect on aggravated and recurrent cutaneous T-cell lymphoma (CTCL), but is accompanied by serious adverse effects. Therefore, oral vorinostat is not applicable to the treatment of early stage CTCL. The aim of this study is to develop a novel vorinostat formulation which is effective for early stage CTCL and free of the serious adverse effects. A mixed-matrix hydrogel of vorinostat was prepared and characterized as a potential topical skin delivery system. Moisture retention, swelling behavior, viscosity, real-time morphology and differential scanning calorimeter analysis (DSC) of hydrogel were evaluated to select the solvent, matrix and humectant. The optimal HPMC/HPC ratio, pH, additive, dose and drug loading of vorinostat hydrogel were determined by evaluating the cumulative vorinostat amount of skin retention and transdermal amount of vorinostat through the skin in vitro. The optimal hydrogel presented a low transdermal amount of vorinostat through the skin, suggesting that the hydrogel reduced the amount of vorinostat that was absorbed in the systemic circulation. More importantly, in vivo percutaneous permeation experiments were also performed to evaluate the permeation behavior of vorinostat into the skin. The topical application with a much lower dose showed higher AUC (the cumulative vorinostat amount of skin retention) than oral application and the hydrogel achieved a sustained permeation of vorinostat in the skin for 24h in vivo. It indicated that a higher relative bioavailability for hydrogel was achieved compared with oral vorinostat. Moreover, there was no damage, inflammation or cell swelling of the skin after administration. Thus, the mixed-matrix vorinostat hydrogel prepared in this study could deliver vorinostat into local skin more efficiently than oral administration.

Design of transparent film-forming hydrogels of tolterodine and their effects on stratum corneum.

A transparent film-forming hydrogel formulation for tolterodine was developed using ternary phase diagram and Box-Behnken design (BBD). Carbopol 980 (neutralized by triethanolamine), hydroxypropyl cellulose (HPC), hydroxypropyl methyl cellulose (HPMC) and Tween 80 were used as matrices. Solvent was the mixture of water and ethyl alcohol. The measured 24 h cumulative drug release rate (86.02%) was consistent with the predicted value (85.42%) in mice. Steady-state flux (J) of tolterodine in optimized formulation across rat full skin, epidermal, dermis and subcutaneous tissue were 15.83, 18.55, 37.15 and 81.82 µg cm(-2) h(-1), respectively. Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) results suggested that the hydrogels could impact lipid status in SC, which was consistent with Ea (8.638 kcal/mol) of tolterodine from optimized formulation in rats. In the pharmacokinetic studies, sustained-release over 24 h and absolute bioavailability of the hydrogels (24.53%) was higher than tolterodine tablets (15.16%) in rats. The hydrogels were suitable for systemic administration of tolterodine for the treatment of overactive bladder.