[Clinical observation on syndrome-differentiation acupuncture combined with rehabilitation training for autism spectrum disorder].

OBJECTIVE: To compare the clinical efficacy and safety between syndrome-differentiation acupuncture combined with rehabilitation training and simple rehabilitation training for children with autism spectrum disorder (ASD). METHODS: A total of 60 children with ASD were randomly divided into an observation group and a control group, 30 cases in each group. In the control group, routine rehabilitation training was applied; in the observation group, syndrome-differentiation acupuncture (the main points were Baihui [GV 20], Dingshenzhen, Niesanzhen, etc., the supplementary acupoints were selected according to syndrome-differentiation) combined with rehabilitation training were applied, all the treatments were given once a day, 5-day continuous treatment with 2-day interval, 12 weeks were required. Before treatment and after 6, 12 weeks of treatment, the autism treatment evaluation checklist (ATEC), childhood autism rating scale (CARS) and autism behavior checklist (ABC) scores were observed, the therapeutic effect and safety were evaluated in the two groups. RESULTS: After 6 and 12 weeks of treatment, except for the sensory perception score after 6 weeks of treatment in the control group, the item scores and total scores of ATEC, CARS scores and ABC scores were decreased compared with those before treatment in the two groups (P<0.05). After 6 weeks of treatment, the social score and total score of ATEC, CARS score in the observation group were lower than those in the control group (P<0.05); after 12 weeks of treatment, the item scores and total score of ATEC, CARS score and ABC score in the observation group were lower than those in the control group (P<0.05). The total effective rate in the observation group was 80.0% (24/30), which was higher than 56.7% (17/30) in the control group (P<0.05). There was no serious adverse reactions in the two groups, and there was no significant difference in the incidence rate of adverse reactions between the two groups (P>0.05). CONCLUSION: Syndrome-differentiation acupuncture combined with rehabilitation training could improve the core symptoms in children with ASD, especially sensory perception and social ability, and with good safety, which is superior to simple rehabilitation training.

Transparent, intrinsically stretchable cellulose nanofiber-mediated conductive hydrogel for strain and humidity sensing.

Conductive hydrogels (CHs) have attracted considerable attentions in the fields of wearable electronics, disease diagnosis, and artificial intelligence. However, it is still a great challenge to prepare a single CH system with integrated characteristics of high stretchability, good transparency, and multisensory function through a simple fabrication process. Herein, carboxylic cellulose nanofibers (CCNF) were used to assist the homogeneous distribution of opaque conductive poly(3,4-ethylenedioxythiophene): poly(styrene sulfonate) (PEDOT: PSS) into the crosslinked polyacrylamide network for the fabrication of stretchable and transparent interpenetrating network CH, aiming for a high-performance multisensory system. As expected, the ready formation of hydrogen bonds between the water molecules and a great deal of hydrophilic groups in the hydrogel endow the obtained CH with excellent humidity response behavior in a wide range (0-85%), and the introduction of CCNF and PEDOT: PSS is proved to be an effective strategy to enhance the humidity sensitivity, exhibiting great potential for the noncontact sensing of human respiration and finger movement. Meanwhile, it also displays excellent strain sensing behavior with favorable sensitivity in a broad range (0-837 %), fast response and reliable stability and reproducibility. Importantly, our prepared CH can also detect and discriminate complicated human activities and physiological signals. All these demonstrate the superiority of our prepared CH for the new generation of flexible wearable electronics.

The Role of Mastering Musical Instrument Playing Skills Combined with Student Behavior Data Mining and Analysis in the Digital Campus Environment to Improve Students' Comprehensive Quality.

Music is closely related to people's lives, and it has a certain impact on people's lives. In school teaching activities, mastering the skills of playing musical instruments can effectively improve students' music appreciation ability and level and enhance students' comprehensive quality through subtle influence. Based on the analysis of students' behavior data, this paper analyzes the role of mastering musical instrument playing skills in improving students' comprehensive quality and puts forward research ideas and schemes. It focuses on students' group behavior in the digital campus environment, integrates multisource data in the digital campus, quantificationally calculates students' multidimensional behaviors, studies the behavior rules of students with different academic performance levels, and uses machine learning algorithm to build a multifeature integrated model of students' comprehensive quality, providing personalized feedback for the improvement of students' comprehensive quality. The results show that the effect of mastering musical instrument playing skills combined with data mining analysis of students' behavior is generally 30% higher than that of the previous research. Compared with a single model, the fused model can fully consider each algorithm to observe data from different data spaces and structures and give full play to the advantages of different algorithms. The training of a single model will fall into the local minimum, which may lead to the relatively poor generalization performance of its model. However, the weighted fusion of multiple basic learners can effectively reduce the probability of falling into the local minimum.

Understanding of supramolecular solution polymerization and interfacial polymerization via forming multiple hydrogen bonds: a computer simulation study.

By employing dissipative particle dynamics (DPD) simulations combined with stochastic polymerization models, we have conducted a detailed simulation study of supramolecular solution polymerization as well as interfacial polymerization employing a coarse-grained model which is closer to the real monomer structure. By adding bending angle potentials to coarse-grained models representing supramolecular reactive monomers, we achieved monomer model simulations for different kinds of multiple hydrogen bonds. Our simulation results indicated that for the interfacial polymerization system, the volume of the monomer caused a strong steric hindrance effect, which in turn led to a low average degree of polymerization of the product. Therefore, by appropriately reducing the volume of the reaction monomer (corresponding to different confinement ascribed to the multiple hydrogen bonds), the average polymerization degree, the degree of reaction and the polymerization rate of the monomer can be effectively improved. For the solution polymerization system and the interfacial polymerization system, a certain proportion of rigid monomers and flexible monomers (60% rigid monomers and 40% flexible monomers) are mixed. High molecular weight products can thus be obtained via the polymerization reaction. The simulation strategy proposed in this study can not only provide theoretical guidance for better design of new supramolecular systems, but also provide ideas for the further synthesis of higher molecular weight supramolecular polymers.

Polymorphonuclear myeloid-derived suppressor cells link inflammation and damage response after trauma.

Elimination of the posttraumatic inflammatory response and recovery of homeostasis are crucial for the positive prognosis of trauma patients. Myeloid-derived suppressor cells (MDSCs) are known to play a regulatory role in the posttraumatic immune response in mice, but their induction source and involved potential mechanism are poorly understood. Here, we report that polymorphonuclear MDSCs (PMN-MDSCs) are activated after trauma and are closely associated with the progression of the posttraumatic inflammatory response. In humans, lectin-type oxidized LDL receptor 1 (LOX1) was used to specifically characterize LOX1+ PMN-MDSCs. Trauma patients showed high intracellular reactive oxygen species (ROS) production, as well as activation of LOX1+ PMN-MDSCs. These MDSCs contribute to the anti-inflammatory immune response by regulating the Treg/Th17 and Th2/Th1 balances after trauma, increasing the levels of anti-inflammatory factors, and decreasing the levels of proinflammatory factors. The number of LOX1+ PMN-MDSCs was positively correlated with the positive clinical prognosis of trauma patients with infection. Activation of LOX1+ PMN-MDSCs is mediated by NF-κB signal, and TGF-ß1 may be as an important inducer for LOX1+ PMN-MDSCs in the posttraumatic cytokine environment. In a pseudofracture trauma mouse model, we also observed the activation of PMN-MDSCs, accompanying high levels of intracellular ROS production, NF-κB phosphorylation, and changes in the inflammatory environment, in particularly by regulating the Treg/Th17 and Th2/Th1 balance. And more significantly, posttraumatic inflammation was alleviated in mice after transferring trauma-derived PMN-MDSCs, but aggravated after injecting with Gr1 agonistic antibody. These findings provide evidence for the specific role of PMN-MDSCs in the regulation of posttraumatic inflammation.

Continuous activation of polymorphonuclear myeloid-derived suppressor cells during pregnancy is critical for fetal development.

The maternal immune system is vital in maintaining immunotolerance to the semiallogeneic fetus for a successful pregnancy. Although studies have shown that myeloid-derived suppressor cells (MDSCs) play an important role in maintaining feto-maternal tolerance, little is known about the role of MDSCs in pregnancies with intrauterine growth retardation (IUGR). Here, we reported that the activation of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) during pregnancy was closely associated with fetal growth. In humans, class E scavenger receptor 1 (SR-E1), a distinct marker for human PMN-MDSCs, was used to investigate PMN-MDSC function during pregnancy. Continuous activation of SR-E1+ PMN-MDSCs was observed in all stages of pregnancy, accompanied by high cellular levels of ROS and arginase-1 activity, mediated through STAT6 signaling. However, SR-E1+ PMN-MDSCs in pregnancies with IUGR showed significantly lower suppressive activity, lower arginase-1 activity and ROS levels, and decreased STAT6 phosphorylation level, which were accompanied by an increase in inflammatory factors, compared with those in normal pregnancies. Moreover, the population of SR-E1+ PMN-MDSCs was negatively correlated with the adverse outcomes of newborns from pregnancies with IUGR. In mice, decreases in cell population, suppressive activity, target expression levels, and STAT6 phosphorylation levels were also observed in the pregnancies with IUGR compared with the normal pregnancies, which were rescued by the adoptive transfer of PMN-MDSCs from pregnant mice. Interestingly, the growth-promoting factors (GPFs) secreted by placental PMN-MDSCs in both humans and mice play a vital role in fetal development. These findings collectively support that PMN-MDSCs have another new role in pregnancy, which can improve adverse neonatal outcomes.

Treatment With Endothelin-A Receptor Antagonist BQ123 Attenuates Acute Inflammation in Mice Through T-Cell-Dependent Polymorphonuclear Myeloid-Derived Suppressor Cell Activation.

The endothelin-A receptor antagonist BQ123 is an effective treatment agent for hypertension and obese cardiomyopathy. However, the role of BQ123 in controlling acute inflammatory diseases and its underlying mechanisms are not well understood. Here, we showed that BQ123 activated polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) in mice and that the IL13/STAT6/Arg1 signaling pathway is involved in this process. Importantly, both treatment with BQ123 and the transfer of BQ123-induced PMN-MDSCs (BQ123-MDSCs) were effective in relieving inflammation, including dextran sulfate sodium (DSS)-induced colitis, papain-induced pneumonia, and concanavalin A (ConA)-induced hepatitis, in mice. The treatment effects were mediated by the attenuation of the inflammation associated with the accumulation of PMN-MDSCs in the colon, lung, and liver. However, concurrent injection of Gr1 agonistic antibody with BQ123 induced PMN-MDSC aggravated the observed acute inflammation. Interestingly, no remission of inflammation was observed in Rag2 knockout mice administered BQ123-MDSCs, but co-injection with CD3+ T cells significantly relieved acute inflammation. In summary, BQ123-induced PMN-MDSCs attenuated acute inflammation in a T cell-dependent manner, providing a novel potential strategy to prevent the occurrence of acute inflammation.

TFF3 mediates the NF-κB/COX2 pathway to regulate PMN-MDSCs activation and protect against necrotizing enterocolitis.

Intestinal trefoil factor 3 (TFF3) plays an important role in repairing the intestinal mucosa. However, the detailed mechanism regarding immune regulation by TFF3 is not well defined. Here, we reported that treatment of mouse BM cells and human peripheral blood mononuclear cells from healthy volunteers with TFF3 activated polymorphnuclear myeloid-derived suppressor cells (PMN-MDSCs) in vitro. We also found that prostaglandin E2 is a major TFF3-mediated MDSC target, and that NF-κB/COX2 signaling was involved in this process. Moreover, TFF3 treatment or transfer of TFF3-derived PMN-MDSCs (TFF3-MDSCs) to experimental necrotizing enterocolitis (NEC) mice caused PMN-MDSC accumulation in the lamina propria (LP), which was associated with decreased intestinal inflammation, permeability, bacterial loading, and prolonged survival. Interestingly, no NEC severity remission was observed in Rag1 KO mice that were given TFF3-MDSCs, but coinjection with CD4+ T cells significantly relieved NEC inflammation. Overall, TFF3 mediates the NF-κB/COX2 pathway to regulate PMN-MDSC activation and attenuates NEC in a T-cell-dependent manner, which suggests a novel mechanism in preventing NEC occurrence.

Safety and efficacy of complementary and alternative medicine in the treatment of autism spectrum disorder: A protocol for systematic review and meta-analysis.

INTRODUCTION: The purpose of this study is to evaluate the efficacy and safety of complementary and alternative medicine in the treatment of autism spectrum disorder. METHODS AND ANALYSIS: We will electronically search Pubmed, Medline, Embase, Web of Science, the Cochrane Central Register of Controlled Trial, China National Knowledge Infrastructure, China Biomedical Literature Database, China Science Journal Database, and Wan-fang Database from their inception. Also, we will manually retrieve other resources, including reference lists of identified publications, conference articles, and gray literature. The clinical randomized controlled trials or quasi-randomized controlled trials related to complementary and alternative medicine treating autism spectrum disorder will be included in the study. The language is limited to Chinese and English. Research selection, data extraction, and research quality assessment will be independently completed by 2 researchers. Data were synthesized by using a fixed-effect model or random-effect model depend on the heterogeneity test. The Childhood Autism Rating Scale (CARS) and Autism Behavior Checklist (ABC) scores will be the primary outcomes. The scores of the Autism Treatment Evaluation Checklist and the Ritvo-Freeman Real Life Rating Scale will also be assessed as secondary outcomes. RevMan V.5.3 statistical software will be used for meta-analysis, and the level of evidence will be assessed by Grading of Recommendations Assessment, Development, and Evaluation (GRADE). Continuous data will be expressed in the form of weighted mean difference or standardized mean difference with 95% confidence intervals (CIs), whereas dichotomous data will be expressed in the form of relative risk with 95% CIs. ETHICS AND DISSEMINATION: The protocol of this systematic review does not require ethical approval because it does not involve humans. We will publish this article in peer-reviewed journals and presented at relevant conferences. SYSTEMATIC REVIEW REGISTRATION: OSF Registries, DOI: 10.17605/OSF.IO/ HA97R (https://osf.io/ha97r).

Identification of linear B cell epitopes on VP1 and VP2 proteins of Senecavirus A (SVA) using monoclonal antibodies.

Senecavirus A (SVA), previously called Seneca Valley virus, belongs to the family Picornaviridae, species Senecavirus A, in the Senecavirus genus, and can cause vesicular lesions in sows and acute death in piglets. In this study, recombinant VP1 and VP2 proteins were expressed in prokaryotic expression system and used to generate eight monoclonal antibodies (mAbs) against VP1 or VP2 protein. And all of the mAbs reacted specifically with SVA virus by both Western blot and indirect immunofluorescence assay (IFA). The resurts showed that all of the epitopes aganist these mAbs were B cell linear epitopes. To map the epitopes, both Western blot and indirect enzyme-linked immunosorbant assay (indirect ELISA) were performed. The epitope 21GELAAP26 recognized by mAb 1G9, was likely to be a significant B cell epitope due to the high antigenic index and the fully exposure on the surface of the VP1. Other mAbs were recognized by VP2 protein. MAbs 1E7 and 8E8 recognized the same epitope at 12DRVITQT18, 1A5 recognized the epitope at 71WTKAVK76, 1G2 recognized the epitope at 98GGAFTA103, 9D2 and 6B11 recognized the same epitope at 150KSLQELN156, and 7E4 recognized the epitope at 248YKEGAT253. Alignment of amino acids revealed that four epitopes were completely conserved among all SVA strains, including 21GELAAP26, 71WTKAVK76, 98GGAFTA103, and 248YKEGAT253. Interestingly, there were some amino acid mutations in 12DRVITQT18 and 150KSLQELN156, but no significant difference was detected on the reaction intensity between epitopes and the corresponding mAbs. This is the first report about the SVA epitopes, which will benefit to the study of viral pathogenic mechanism, vaccine design, as well as the establishment of detection methods.

Cholesterol 25-hydroxylase inhibits encephalomyocarditis virus replication through enzyme activity-dependent and independent mechanisms.

Cholesterol-25-hydroxylase (CH25 H) is a reticulum-associated membrane protein induced by an important interferon-stimulating gene (ISG) and can significantly inhibit some virus replication. But the effect of CH25H on encephalomyocarditis virus (EMCV) is still not clear. In this study, we found that EMCV infection increases significantly the endogenous CH25H expression in BHK-21 and N2a cells. CH25H and cholesterol catalytic oxidation product 25-hydroxycholesterol (25HC) obviously inhibits EMCV infection by inhibiting the viral penetration. But the CH25H mutant lacking hydroxylase activity repairs the ability to inhibit the viral replication. Meanwhile, ß-cyclodextrin crystalline as a cholesterol inhibitor significantly decreases the viral replication. In addition, CH25H can selectively interact and degrade the viral RNA-Dependent RNA Polymerase-3D protein by independent on the association of proteasome, lysosome and caspase manner. It provides new insights into the interplay mechanisms between CH25H and non-enveloped single-stranded positive RNA viruses.

Inhibition of encephalomyocarditis virus replication by shRNA targeting 1C and 2A genes in vitro and in vivo.

Encephalomyocarditis virus (EMCV) infects many mammalian species, causing myocarditis, encephalitis and reproductive disorders. The small interference RNA (siRNA) targeting to the virus has not been understood completely. Here, two out of six interference sequences were screened to inhibit significantly EMCV replication by using recombinant plasmids expressing small hairpin RNA (shRNA) targeting to the viral 1C or 2A genes in BHK-21 cells. And two recombinant adenoviruses expressing the shRNAs were constructed and named as rAd-1C-1 and rAd-2A-3. They inhibit EMCV replication in BHK-21 cells in protein levels, as well as the virus yields by approximately 1000 times. Furthermore, they provide high protective efficacy against the challenge with virulent EMCV NJ08 strain in mice. And the EMCV loads in the live mice in rAd-1C-1 and rAd-2A-3 groups decrease by more than 90 % compared with those in the dead mice in the challenge control groups at the same times. It indicates that the adenoviruses medicated shRNA targeting to 1C and 2A genes might provide a potential strategy for combating EMCV infection.

Characterization and functional analysis of chicken APOBEC4.

The APOBEC proteins play significant roles in the innate and adaptive immune system, probably due to their deaminase activities. Because APOBEC1 (A1) and APOBEC3 (A3) are absent in the chicken genome, we were interested in determining whether chicken APOBEC4 (A4) possessed more complex functions than its mammalian homologs. In this study, chicken A4 (chA4) mRNA was identified and cloned for the first time. Based on bioinformatics analyses, the conserved zinc-coordinating motif (HXE … PC(X)2-6C) was identified on the surface of chA4 and contained highly conserved His97, Glu99, Pro130, Cys131 and Cys138 active sites. The highest expression levels of constitutive chA4 were detected in primary lymphocytes and bursa of Fabricius. Newcastle Disease (ND) is one of the most serious infectious diseases in birds, causing major economic losses to the poultry industry. In vitro, Newcastle Disease Virus (NDV) early infection induced significant increases in chA4 expression in the chicken B cell line, DT40, the macrophage cell line, HD11 and the CD4+ T cell line, MSB-1, but not the fibroblast cell line, DF-1. In vivo, the expression levels of chA4 were up-regulated in several tissues from NDV-infected chickens, especially the thymus, testicles, duodenum and kidney. The high level expression of exogenous chA4 displayed inhibitory effects on NDV and reduced viral RNA in infected cells. Taken together, these data demonstrate that chA4 is involved in the chicken immune system and may play important roles in host anti-viral responses.

Hemagglutinin-neuraminidase and fusion proteins of virulent Newcastle disease virus cooperatively disturb fusion-fission homeostasis to enhance mitochondrial function by activating the unfolded protein response of endoplasmic reticulum and mitochondrial stress.

The fusogenically activated F and HN proteins of virulent NDV induce complete autophagic flux in DF-1 and A549 cells. However, the effect of both glycoproteins on mitochondria remains elusive. Here, we found that F and HN cooperation increases mitochondrial biogenesis but does not cause the mitochondria damage. We observed that both glycoproteins change the morphological characteristics and spatial distribution of intracellular mitochondria. F and HN cooperate cooperatively to induce ER stress and UPRmt. Our preliminary data suggested that F and HN cooperatively disturb mitochondrial fusion-fission homeostasis to enhance mitochondrial biogenesis, and eventually meet the energy demand of syncytium formation.

Syncytia generated by hemagglutinin-neuraminidase and fusion proteins of virulent Newcastle disease virus induce complete autophagy by activating AMPK-mTORC1-ULK1 signaling>.

Autophagy triggered by glycoprotein-mediated membrane fusion has been reported for several paramyxoviruses. However, the function of HN and F glycoproteins of NDV and their role in autophagy induction have not been studied. Here, we found that co-transfection of HN and F of virulent NDV rapidly induced syncytium formation and triggered a steady state autophagy flux in adenocarcinomic human alveolar basal epithelial (A549) cells and chicken embryo fibroblast (DF-1) cells. Furthermore, we clearly identified that F and HN synergistically induced autophagosome fusion with lysosomes for subsequent degradation. The seven cleavage site mutations of F significantly decreased the autophagy induction, compared with those of wildtype virulent F. RNAi and pharmacological experiments suggested that autophagy benefitted membrane fusion and syncytium formation induced by F and HN of NDV. Activated F1 co-operated with HN to stimulate AMPK kinase and downstream ULK1 activation to suppress mTORC1 signaling. Our data described the synergistic role of HN and F in the induction of completed autophagic flux through the activation of AMPK- mTORC1- ULK1 pathway.

A node influence based label propagation algorithm for community detection in networks.

Label propagation algorithm (LPA) is an extremely fast community detection method and is widely used in large scale networks. In spite of the advantages of LPA, the issue of its poor stability has not yet been well addressed. We propose a novel node influence based label propagation algorithm for community detection (NIBLPA), which improves the performance of LPA by improving the node orders of label updating and the mechanism of label choosing when more than one label is contained by the maximum number of nodes. NIBLPA can get more stable results than LPA since it avoids the complete randomness of LPA. The experimental results on both synthetic and real networks demonstrate that NIBLPA maintains the efficiency of the traditional LPA algorithm, and, at the same time, it has a superior performance to some representative methods.