Foot-and-Mouth Disease Virus 3B Protein Interacts with Pattern Recognition Receptor RIG-I to Block RIG-I-Mediated Immune Signaling and Inhibit Host Antiviral Response.

Foot-and-mouth disease is a highly contagious disease of pigs, sheep, goats, bovine, and various wild cloven-hoofed animals caused by foot-and-mouth disease virus (FMDV) that has given rise to significant economic loss to global livestock industry. FMDV 3B protein is an important determinant of virulence of the virus. Modifications in 3B protein of FMDV considerably decrease virus yield. In the current study, we demonstrated the significant role of 3B protein in suppression of type I IFN production and host antiviral response in both human embryonic kidney HEK293T cells and porcine kidney PK-15 cells. We found that 3B protein interacted with the viral RNA sensor RIG-I to block RIG-I-mediated immune signaling. 3B protein did not affect the expression of RIG-I but interacted with RIG-I to block the interaction between RIG-I and the E3 ubiquitin ligase TRIM25, which prevented the TRIM25-mediated, Lys63-linked ubiquitination and activation of RIG-I. This inhibition of RIG-I-mediated immune signaling by 3B protein decreased IFN-ß, IFN-stimulated genes, and proinflammatory cytokines expression, which in turn promoted FMDV replication. All of the three nonidentical copies of 3B could inhibit type I IFN production, and the aa 17A in each copy of 3B was involved in suppression of IFN-related antiviral response during FMDV infection in porcine cells. Together, our results indicate the role of 3B in suppression of host innate immune response and reveal a novel antagonistic mechanism of FMDV that is mediated by 3B protein.

Preparation of Silica Aerogel/Resin Composites and Their Application in Dental Restorative Materials.

As the most advanced aerogel material, silica aerogel has had transformative industrial impacts. However, the use of silica aerogel is currently limited to the field of thermal insulation materials, so it is urgent to expand its application into other fields. In this work, silica aerogel/resin composites were successfully prepared by combining silica aerogel with a resin matrix for dental restoration. The applications of this material in the field of dental restoration, as well as its performance, are discussed in depth. It was demonstrated that, when the ratio of the resin matrix Bis-GMA to TEGDMA was 1:1, and the content of silica aerogel with 50 µm particle size was 12.5%, the composite achieved excellent mechanical properties. The flexural strength of the silica aerogel/resin composite reached 62.9546 MPa, which was more than five times that of the pure resin. Due to the presence of the silica aerogel, the composite also demonstrated outstanding antibacterial capabilities, meeting the demand for antimicrobial properties in dental materials. This work successfully investigated the prospect of using commercially available silica aerogels in dental restorative materials; we provide an easy method for using silica aerogels as dental restorative materials, as well as a reference for their application in the field of biomedical materials.

Green Preparation of Durian Rind-Based Cellulose Nanofiber and Its Application in Aerogel.

In this study, a green, highly efficient and low energy consumption preparation method of cellulose nanofiber (CNF) was developed by using agricultural and forestry waste durian rinds as raw materials. The power of ultrasonic treatment was successfully reduced to only 360 W with low molecular weight liquid DMSO. The obtained durian rind-based CNF had a diameter of 8-20 nm and a length of several micrometers. It had good dispersion and stability in water, and could spontaneously cross-link to form hydrogel at room temperature when the concentration was more than 0.5%. The microscopic morphology and compressive properties of CNF aerogels and composite cellulose aerogels prepared from durian rind-based CNF were evaluated. It was found that CNF could effectively prevent the volume shrinkage of aerogel, and the concentration of CNF had a significant effect on the microstructure and mechanical properties of aerogel. The CNF aerogel with 1% CNF exhibited a sheet structure braced by fibers, which had the strongest compression performance. The porosity of CNF aerogels was high to 99%. The compressive strength of the composite cellulose aerogel with durian rind-based CNF was effectively enhanced.

Foot-and-Mouth Disease Virus Capsid Protein VP1 Interacts with Host Ribosomal Protein SA To Maintain Activation of the MAPK Signal Pathway and Promote Virus Replication.

Foot-and-mouth disease virus (FMDV) is the causative agent of foot-and-mouth disease, a highly contagious, economically important viral disease. The structural protein VP1 plays significant roles during FMDV infection. Here, we identified that VP1 interacted with host ribosomal protein SA (RPSA). RPSA is a viral receptor for dengue virus and classical swine fever virus infections. However, the incubation of susceptible cells using the anti-RPSA antibodies did not block the infection of FMDV. Overexpression of porcine RPSA in the insusceptible cells could not trigger FMDV infection, suggesting that RPSA was not responsible for FMDV entry and infection. On the contrary, we found that overexpression of RPSA suppressed FMDV replication, and knockdown of RPSA enhanced FMDV replication. We further determined that FMDV infection activated the mitogen-activated protein kinase (MAPK) pathway and demonstrated that MAPK pathway activation was critically important for FMDV replication. RPSA negatively regulated MAPK pathway activation during FMDV infection and displayed an antiviral function. FMDV VP1 interacted with RPSA to abrogate the RPSA-mediated suppressive role in MAPK pathway activation. Together, our study indicated that MAPK pathway activation was required for FMDV replication and that host RPSA played a negatively regulatory role on MAPK pathway activation to suppress FMDV replication. FMDV VP1 bound to RPSA to promote viral replication by repressing RPSA-mediated function and maintaining the activation of MAPK signal pathway.IMPORTANCE Identification of virus-cell interactions is essential for making strategies to limit virus replication and refine the models of virus replication. This study demonstrated that FMDV utilized the MAPK pathway for viral replication. The host RPSA protein inhibited FMDV replication by suppressing the activation of the MAPK pathway during FMDV infection. FMDV VP1 bound to RPSA to repress the RPSA-mediated regulatory effect on MAPK pathway activation. This study revealed an important implication of the MAPK pathway for FMDV infection and identified a novel mechanism by which FMDV VP1 has evolved to interact with RPSA and maintain the activation of the MAPK pathway, elucidating new information regarding the signal reprogramming of host cells by FMDV.

Clinical features of obstructive sleep apnea in children with obesity. / è‚¥èƒ–å„¿ç«¥é˜»å¡žæ€§ç¡çœ å‘¼å¸æš‚åœçš„ä¸´åºŠç‰¹å¾.

OBJECTIVES: To study the clinical features of obstructive sleep apnea (OSA) in children with obesity. METHODS: A retrospective analysis was performed on the medical data of 33 obese children aged 7-15 years, who were diagnosed with OSA and received polysomnography (PSG) in the Department of Respiratory Medicine in Shenzhen Children's Hospital. Fifty OSA children with normal body weight, matched for sex and age, were enrolled as the control group. RESULTS: Among the 33 obese children with OSA, the three most common daytime symptoms were inattention in 30 children (91%), somnolence in 22 children (67%), and morning fatigue in 21 children (64%), and the three most common nocturnal symptoms were snoring in 27 children (82%), mouth breathing in 20 children (61%), and sweating in 16 children (49%). Compared with the reference values of normal children, both the OSA + obesity group and the control group had prolonged light sleep, shortened deep sleep, and a significantly shortened rapid eye movement (REM) period, while there was no significant difference in these indices between the two groups (P>0.05). The apnea-hypopnea index, obstructive apnea/hypopnea index, and oxygen desaturation index in both REM and non-REM periods in the OSA +obesity group were significantly higher than those in the control group (P<0.05), while the lowest blood oxygen saturation during sleep was significantly lower in the OSA + obesity group (P<0.05). CONCLUSIONS: The children with obesity and OSA have the main daytime symptoms of inattention, somnolence, and morning fatigue and the main nocturnal symptoms of snoring, mouth breathing, and sweating. There is no significant difference in sleep structure between OSA children with obesity and those with normal body weight; however, respiratory events and blood oxygen saturation decline are more severe in OSA children with obesity. Citation.

The Cotton BEL1-Like Transcription Factor GhBLH7-D06 Negatively Regulates the Defense Response against Verticillium dahliae.

Verticillium wilt will seriously affect cotton yield and fiber quality. BEL1-Like transcription factors are involved in the regulation of secondary cell wall (SCW) formation, especially the biosynthesis of lignin that also plays a key role in cotton disease resistance. However, there is no report on the role of BEL1-Like transcription factor in the regulation of plant biological stress. In this study, tissue expression pattern analysis showed that a BEL1-Like transcription factor GhBLH7-D06 was predominantly expressed in vascular tissues and the SCW thickening stage of fiber development, while its expression could also respond to Verticillium dahliae infection and the phytohormone MeJA treatment, which indicated that GhBLH7-D06 might be involved in the defense response of Verticillium wilt. Using virus-induced gene silencing (VIGS) technology, we found silencing the expression of GhBLH7-D06 could enhance the resistance of cotton plants to Verticillium wilt, and the acquisition of resistance might be mainly due to the significant overexpression of genes related to lignin biosynthesis and JA signaling pathway, which also proves that GhBLH7-D06 negatively regulates the resistance of cotton to Verticillium wilt. Based on the results of yeast two-hybrid (Y2H) library screening and confirmation by bimolecular fluorescence complementary (BiFC) experiment, we found an Ovate Family Protein (OFP) transcription factor GhOFP3-D13 which was also a negative regulator of cotton Verticillium wilt resistance could that interacts with GhBLH7-D06. Furthermore, the dual-luciferase reporter assay and yeast one-hybrid (Y1H) experiment indicated that GhBLH7-D06 could target binding to the promoter region of GhPAL-A06 to suppress its expression and eventually lead to the inhibition of lignin biosynthesis. In general, the GhBLH7-D06/GhOFP3-D13 complex can negatively regulate resistance to Verticillium wilt of cotton by inhibiting lignin biosynthesis and JA signaling pathway.

Genome-wide identification and characterization of TALE superfamily genes in cotton reveals their functions in regulating secondary cell wall biosynthesis.

BACKGROUND: Cotton fiber length and strength are both key traits of fiber quality, and fiber strength (FS) is tightly correlated with secondary cell wall (SCW) biosynthesis. The three-amino-acid-loop-extension (TALE) superclass homeoproteins are involved in regulating diverse biological processes in plants, and some TALE members has been identified to play a key role in regulating SCW formation. However, little is known about the functions of TALE members in cotton (Gossypium spp.). RESULTS: In the present study, based on gene homology, 46, 47, 88 and 94 TALE superfamily genes were identified in G. arboreum, G. raimondii, G. barbadense and G. hirsutum, respectively. Phylogenetic and evolutionary analysis showed the evolutionary conservation of two cotton TALE families (including BEL1-like and KNOX families). Gene structure analysis also indicated the conservation of GhTALE members under selection. The analysis of promoter cis-elements and expression patterns suggested potential transcriptional regulation functions in fiber SCW biosynthesis and responses to some phytohormones for GhTALE proteins. Genome-wide analysis of colocalization of TALE transcription factors with SCW-related QTLs revealed that some BEL1-like genes and KNAT7 homologs may participate in the regulation of cotton fiber strength formation. Overexpression of GhKNAT7-A03 and GhBLH6-A13 significantly inhibited the synthesis of lignocellulose in interfascicular fibers of Arabidopsis. Yeast two-hybrid (Y2H) experiments showed extensive heteromeric interactions between GhKNAT7 homologs and some GhBEL1-like proteins. Yeast one-hybrid (Y1H) experiments identified the upstream GhMYB46 binding sites in the promoter region of GhTALE members and defined the downstream genes that can be directly bound and regulated by GhTALE heterodimers. CONCLUSION: We comprehensively identified TALE superfamily genes in cotton. Some GhTALE members are predominantly expressed during the cotton fiber SCW thicking stage, and may genetically correlated with the formation of FS. Class II KNOX member GhKNAT7 can interact with some GhBEL1-like members to form the heterodimers to regulate the downstream targets, and this regulatory relationship is partially conserved with Arabidopsis. In summary, this study provides important clues for further elucidating the functions of TALE genes in regulating cotton growth and development, especially in the fiber SCW biosynthesis network, and it also contributes genetic resources to the improvement of cotton fiber quality.

Effects of glycine 64 substitutions in RNA-dependent RNA polymerase on ribavirin sensitivity and pathogenicity of coxsackievirus A6.

Hand, foot, and mouth disease (HFMD) caused by a group of enteroviruses is a global public health problem. In recent years, coxsackievirus A6 (CVA6) has emerged as an important HFMD agent. Previous studies have shown that mutations of glycine 64 in RNA-dependent RNA polymerase (3D polymerase), which is central to viral replication, cause phenotypic changes such as ribavirin resistance, increased replication fidelity, and virulence attenuation in poliovirus and enterovirus A71. In this study, we constructed CVA6 mutants with G64R, G64S, and G64T substitutions by site-directed mutagenesis in full-length cDNA of an infectious CVA6 strain cloned in pcDNA3.1. Viral RNA was obtained by in vitro transcription, and the rescued virus strains were propagated in RD cells. Sequencing after six passages revealed that G64S and G64T mutations were stably inherited, whereas G64R was genetically unstable and reversed to the wild type. Comparison of the biological characteristics of the wild-type and mutant CVA6 strains in an in vivo model (one-day-old ICR mice) revealed that the pathogenicity of CVA6-G64S and CVA6-G64T was significantly reduced compared to wild-type CVA6. In vitro experiments indicated the mutant CVA6-G64S and CVA6-G64T strains had increased resistance to 0.8 mM ribavirin and a decreased replication rate in the presence of 0.8 mM guanidine hydrochloride. Our results show that mutation of residue 64 reduces CVA6 susceptibility to ribavirin and increases CVA6 susceptibility to guanidine hydrochloride, together with increased replication fidelity and attenuated viral pathogenicity, thus laying a foundation for the development of safe and effective live attenuated CVA6 vaccine.

A novel report on the distribution of microplastics in ocellated icefish (Chionodraco rastrospinosus) in the northern Antarctic Peninsula.

Plastics are ubiquitous in the global marine ecosystem; however, studies on microplastic (MPs) ingestion by fish in the remote Antarctic are extremely limited. We detected MPs in ocellated icefish (Chionodraco rastrospinosus)-a limitedly distributed but ecologically important species-in the northern Antarctic Peninsula. This is the first study reporting MPs distribution in the stomach, intestine, and gill of this species. Of 32 C. rastrospinosus individuals, 10 (31.3 %) individuals ingested MPs. Totally, 157 suspected microplastic particles were detected, and only 10 MPs were finally confirmed from the subsampled 38 highly suspected particles after FTIR detection, with an average of 0.36 particles per individual. The stomach, intestine, and gills of each fish contained 0.06 ± 0.24, 0.16 ± 0.36, and 0.09 ± 0.29 particles, respectively. Fibrous MPs with a diameter of <200 µm were predominant in all tissues, with blue MPs being the most common. Polyester and acrylic acid were the most frequently observed plastic polymers. MPs abundance in C. rastrospinosus was unrelated to biometrics.

Toxic effects of microplastics and nanoplastics on plants: A global meta-analysis.

Microplastics (MPs) and nanoplastics (NPs) pervade and accumulate within the soil, exerting influences not only on plant growth and development but also on human health through the food chain. However, recent studies on the effects of MPs/NPs on plants yield diverse results. Thus, this study conducts a meta-analysis of 101 recent publications to summarize the influence of type, size, and concentration of MPs/NPs on physiological responses, photosynthetic pigments, and biochemical reactions in various plant species. The findings indicate that the effects of MPs/NPs on plants vary depending on the specific plant species. The impact of different polymer types of MPs/NPs on plants does not show a clear trend, possibly due to variations in polymer functional groups. However, it is noteworthy that polyethylene terephthalate with phenyl groups exhibits the most significant effect on plant fresh weight, chlorophyll a and b, and H2O2 content among all microplastic types. Moreover, MPs exhibit stronger inhibition on most physiological, photosynthetic pigments, and biochemical indicators of plants compared to NPs, although root length, chlorophyll, and H2O2 demonstrate opposite effects. The concentration of MPs/NPs elicits different responses on plant indicators, each with specific trends. Notably, exposure to MPs/NPs stimulates an increase in plant biochemical enzyme indicators. Finally, this study identifies current knowledge gaps and offers insights for future research directions.

Coxsackievirus A6 Infection Causes Neurogenic Pathogenesis in a Neonatal Murine Model.

Coxsackievirus A6 (CVA6), a member of species A enterovirus, is associated with outbreaks of hand-foot-and-mouth disease and causes a large nationwide burden of disease. However, the molecular pathogenesis of CVA6 remains unclear. In the present study, we established a suckling Institute of Cancer Research (ICR) mouse infection model to explore the neural pathogenicity of CVA6. Five-day-old mice infected with CVA6 strain F219 showed lethargy and paralysis, and died 5 or 6 days after infection via IM injection. Cerebral edema and neuronal cell swelling were observed in the infected brain tissue, and we found that the CVA6 VP1 antigen could co-localize with GFAP-positive astrocytes in infected mouse brain using an immunofluorescence assay. CVA6 strain F219 can also infect human glioma (U251) cells. Transcriptome analysis of brain tissues from infected mice and infected U251 cells showed that significantly differentially expressed genes were enriched in antiviral and immune response and neurological system processes. These results indicate that CVA6 could cause neural pathogenesis and provide basic data for exploring the mechanism of how host-cell interactions affect viral replication and pathogenesis. Importance: Coxsackievirus A6 (CVA6) surpasses the two main pathogens, enterovirus 71 (EV-A71) and coxsackievirus A16 (CVA16), which are the leading pathogens causing HFMD in many provinces of China. In our study, CVA6 infection caused neurogenic pathogenesis in a neonatal murine model, manifesting as cerebral edema and neuronal cell swelling, CVA6 VP1 antigen could co-localize with GFAP-positive astrocytes in the infected mouse brain. Based on CVA6-infected brain tissue and U251 cell transcriptome analysis, we found upregulated antiviral and immune response-related genes such as Zbp1, Usp18, Oas2, Irf7, Ddx60, Ifit3, Ddx58, and Isg15, while the neurological system process-related genes were downregulated, including Fcrls, Ebnrb, Cdk1, and Anxa5.

Transcriptome sequencing analysis of echovirus 30 infection reveals its potential pathogenesis.

Echovirus 30 (E30) causes various diseases, such as viral encephalitis; aseptic meningitis; hand, foot, and mouth diseases; and acute flaccid paralysis. Related neurological infections are most concerning. However, the molecular mechanisms of E30 pathogenesis are not fully understood. There is a growing research interest in E30 as a cause of neurological disease. The aim of this study was to describe E30 infection, especially the changes in differential factor expressions after infection, in human glioma (U251) cells and mice brains using transcriptome sequencing analysis. Clear changes in the gene expression of factors associated with the defense response to viruses, inflammation-related signaling pathways, and neurological complication-related pathways were observed. Our results suggest that after E30 infection, the genes related to immune response were induced in the human glioma cells and mice brains, whereas genes functioning in the development and function of neural tissue were inhibited. Overall, this study successfully established E30 infection of U251 and mouse brain tissue, profiled the infection-induced changes in cellular and organizational transcriptomes, and revealed the molecular level changes during E30 infection.

Analysis of Thermal Insulation Thickness for a Container House in the Yanqing Zone of the Beijing 2022 Olympic and Paralympic Winter Games.

A large number of temporary housings (THs) were used in the Yanqing zone of the Beijing 2022 Olympic and Paralympic Winter Games. Taking a kind of container house (CH) used in Yanqing zone as a model, the objective of this paper is to analyze the effect of insulation thickness on heating energy consumption and corresponding carbon emission. The effect of service life of THs on economic thickness was also discussed. The simulation model was developed using EnergyPlus and the heating energy consumption with different insulation materials was simulated based on the meteorological parameters of the top of Xiaohaituo Mountain (2177.5 m) and the Olympic/Paralympic Village (950 m) in Yanqing zone. In the simulation process, the thermal insulation performance of the CH was enhanced with reference to the requirements of GB/T 51350-2019 Technical Standard for Nearly Zero Energy Buildings (NZEB) on one hand. Additionally, the insulation performance was evaluated in terms of payback period and carbon emission. On the other hand, the economic thickness of different insulation materials (rock wool (RW), extruded polystyrene (XPS), polyurethane (PU)) and the high performance vacuum insulation panel (HVIP)) for different service lives of CH was studied. Results show that the U-values of the envelope meeting the NZEB standard can decrease approximately 21.4-32.8% of the heating energy consumption, compared with the original envelope. When the service life of CH is extended to 20 years, the carbon emission is reduced by 18.5% and 29.5%. The payback period of HVIP is longest, up to 31.4 a, and the results of economic thickness show that when the service life of the CH ranges from 1 year to 20 years, the economic thickness range of RW is 47-235 mm, XPS is 41-197 mm, PU is 33-149 mm and HVIP is 4-18 mm at the altitude of 2177.5 m. At the altitude of 950 m, the economic thickness range of RW is 28-158 mm, XPS is 26-131 mm, PU is 25-118 mm, and HVIP is 2-12 mm. From an economic point of view, the service life of a CH has a significant impact on the choice of insulation thickness.

Virus-Host Interactions in Foot-and-Mouth Disease Virus Infection.

Foot-and-mouth disease (FMD) is a highly contagious disease of cloven-hoofed animals, which has been regarded as a persistent challenge for the livestock industry in many countries. Foot-and-mouth disease virus (FMDV) is the etiological agent of FMD that can spread rapidly by direct and indirect transmission. FMDV is internalized into host cell by the interaction between FMDV capsid proteins and cellular receptors. When the virus invades into the cells, the host antiviral system is quickly activated to suppress the replication of the virus and remove the virus. To retain fitness and host adaptation, various viruses have evolved multiple elegant strategies to manipulate host machine and circumvent the host antiviral responses. Therefore, identification of virus-host interactions is critical for understanding the host defense against virus infections and the pathogenesis of the viral infectious diseases. This review elaborates on the virus-host interactions during FMDV infection to summarize the pathogenic mechanisms of FMD, and we hope it can provide insights for designing effective vaccines or drugs to prevent and control the spread of FMD and other diseases caused by picornaviruses.

Modification of Metal-Organic Framework Nanoparticles Using Dental Pulp Mesenchymal Stem Cell Membranes to Target Oral Squamous Cell Carcinoma.

Engineering a targetable nanoparticle to tumor cell is a challenge issue for clinical application. Our results demonstrated that the chemokine CXCL8 secreted by oral squamous cell carcinoma (OSCC) could act as a chemoattractant to attract dental pulp mesenchymal stem cell (DPSC), which expressed the CXCL8 binding receptor, CXCR2, to the OSCC. Therefore, to create OSCC targetable nanoparticles, we used DPSC membranes to modify nanoparticles of metal-organic framework nanoparticles (MOFs) resulting in a novel MOF@DPSCM nanoparticle. Interestingly, results from in vitro and in vivo experiments illustrated that MOF@DPSCM possessed specificity for the OSCC, and the MOF@DPSCM carried DOX (doxorubicin), MOF-DOX@DPSCM could induce CAL27 cell death in vitro and block CAL27 tumor growth in vivo. Our data suggest that this novel MOF-DOX@DPSCM nanoparticle is a potential targetable drug delivery system for the OSCC in the future clinical application.

Porcine Picornavirus 3C Protease Degrades PRDX6 to Impair PRDX6-mediated Antiviral Function.

Peroxiredoxin-6 (PRDX6) is an antioxidant enzyme with both the activities of peroxidase and phospholipase A2 (PLA2), which is involved in regulation of many cellular reactions. However, the function of PRDX6 during virus infection remains unknown. In this study, we found that the abundance of PRDX6 protein was dramatically decreased in foot-and-mouth disease virus (FMDV) infected cells. Overexpression of PRDX6 inhibited FMDV replication. In contrast, knockdown of PRDX6 expression promoted FMDV replication, suggesting an antiviral role of PRDX6. To explore whether the activity of peroxidase and PLA2 was associated with PRDX6-mediated antiviral function, a specific inhibitor of PLA2 (MJ33) and a specific inhibitor of peroxidase activity (mercaptosuccinate) were used to treat the cells before FMDV infection. The results showed that incubation of MJ33 but not mercaptosuccinate promoted FMDV replication. Meanwhile, overexpression of PRDX6 slightly enhanced type I interferon signaling. We further determined that the viral 3Cpro was responsible for degradation of PRDX6, and 3Cpro-induced reduction of PRDX6 was independent of the proteasome, lysosome, and caspase pathways. The protease activity of 3Cpro was required for induction of PRDX6 reduction. Besides, PRDX6 suppressed the replication of another porcine picornavirus Senecavirus A (SVA), and the 3Cpro of SVA induced the reduction of PRDX6 through its proteolytic activity as well. Together, our results suggested that PRDX6 plays an important antiviral role during porcine picornavirus infection, and the viral 3Cpro induces the degradation of PRDX6 to overcome PRDX6-mediated antiviral function.

Interleukin-7 promotes CD8+ T cell activity in patients with enterovirus 71 associated encephalitis.

Enterovirus 71 (EV71) always induces severe hand, foot, and mouth disease with neurological complications, such as encephalitis. Interleukin (IL)-7 augments CD8+ T cells activity in chronic viral infection and cancers. However, few studies have focused on common γ-chain (γc) cytokine expression and regulatory function of IL-7 to CD8+ T cells in EV71 associated encephalitis. In this study, twenty-one patients with EV71 associated encephalitis, twenty-seven patients with febrile convulsion (FC), and twenty healthy individuals were enrolled. γc cytokine (IL-2, IL-4, IL-7 and IL-15) concentration was measured by ELISA. IL-7 receptor α chain (membrane/soluble CD127) expression was also investigated. Purified CD8+ T cells were stimulated with recombinant human IL-7 in vitro. The regulatory activity of IL-7 to CD8+ T cells from peripheral blood and cerebrospinal fluids (CSF) was investigated in direct and indirect contact co-culture with U-87MG cells. IL-7 in the serum and CSF, but not IL-2, IL-4, or IL-15, was significant increased in EV71 associated encephalitis. Both total CD127 mRNA relative level and membrane/soluble CD127 expression was comparable among three groups. IL-7 stimulation promoted CD8+ T cells proliferation, up-regulated perforin/granzyme B level, but reduced programmed death-1 expression in CD8+ T cells from EV71 associated encephalitis patients. Cytotoxicity and interferon-γ production of CD8+ T cells from peripheral blood and CSF was also augmented in response to IL-7 stimulation in both direct and indirect co-culture systems in EV71 associated encephalitis. The present data indicated that IL-7 induced cytolytic and non-cytolytic functions of CD8+ T cells in EV71 associated encephalitis. IL-7 might be considered as one of the immunomodulatory therapeutic candidates for EV71 infection.

BMSC affinity peptide-functionalized ß-tricalcium phosphate scaffolds promoting repair of osteonecrosis of the femoral head.

BACKGROUND: Osteonecrosis of the femoral head (ONFH) is a disabling disease. Early treatment is crucial to the prognosis of the disease. Core decompression (CD) is one of the most commonly used methods for the treatment of early ONFH. But it could not prevent the collapse of the necrotic femoral head. How to improve the therapeutic effect of early ONFH on the basis of CD has become an area of focused research. METHODS: Functional ß-tricalcium phosphate (ß-TCP) scaffolds modified by DPIYALSWSGMA (DPI) peptide, a bone marrow-derived mesenchymal stem cell (BMSC) affinity peptide, were constructed using an adsorption/freeze-drying strategy. The affinity of DPI peptide towards rabbit BMSCs was investigated using flow cytometry and fluorescence cytochemistry. In vitro cell adhesion assay was performed to study the adherent ability of rabbit BMSCs on functional ß-TCP scaffolds. After the rabbit model of early ONFH was established, DPI peptide-modified and pure ß-TCP scaffolds were transplanted into the remaining cavity after CD. Meanwhile, rabbits treated with pure CD were used as blank control. Twelve weeks after surgery, histological analysis was performed to show the therapeutic effect of three methods on early ONFH. RESULTS: The result of ImageXpress Micro Confocal indicated that fabricated DPI peptide-modified functional ß-TCP scaffolds exhibited green fluorescence. In flow cytometry, the average fluorescence intensity for rabbit BMSCs incubated with FITC-DPI was significantly higher than that of FITC-LSP (P = 2.733 × 10-8). In fluorescence cytochemistry, strong fluorescent signals were observed in rabbit BMSCs incubated with FITC-DPI and FITC-RGD, whereas no fluorescent signals in cells incubated with FITC-LSP. In cell adhesion assay, the number of adherent cells to ß-TCP-DPI scaffolds was more than that of pure ß-TCP scaffolds (P = 0.033). The CD + ß-TCP-DPI group expressed the lowest vacant bone lacunae percentage compared to CD group (P = 2.350 × 10-4) and CD + ß-TCP group (P = 0.020). The expression content of COL1 in CD + ß-TCP-DPI group was much higher than CD group (P = 1.262 × 10-7) and CD + ß-TCP group (P = 1.666 × 10-7) according to the integrated optical density (IOD) analyses. CONCLUSION: Functional ß-TCP scaffolds modified by DPI peptide were successfully synthesized using an adsorption/freeze-drying strategy. DPI peptide has good affinity towards rabbit BMSCs. The adhesion of rabbit BMSCs on DPI peptide-modified ß-TCP scaffolds was apparently enhanced. CD followed by implantation of DPI peptide-modified ß-TCP scaffolds can apparently improve the treatment of early ONFH compared with pure CD and CD followed by implantation of unmodified ß-TCP scaffolds. Our current study provides an improved method for the treatment of early ONFH.

ß-Arrestin-1 directly interacts with Gαs and regulates its function.

ß-Arrestins function to mediate G protein-coupled receptor (GPCR) desensitization and internalization and to initiate G protein independent signaling of GPCRs. Elucidating how ß-arrestin and G protein signal pathways coordinate with each other is important to fully understand GPCR signaling. Here we report that ß-arrestin-1 directly interacts with Gα(s). Purified ß-arrestin-1 binds to Gα(s) in a rapid association and dissociation manner. ß-Arrestin-1 promotes the binding and the release of GTPγS from Gα(s) in vitro. ß-Arrestin-1 L33K mutant shows reduced interaction with Gα(s) and has no detectable effects on Gα(s) function. Our study thus reveals a direct crosstalk of ß-arrestin-1 with Gα(s).