TRIM33 plays a critical role in regulating dendritic cell differentiation and homeostasis by modulating Irf8 and Bcl2l11 transcription.

The development of distinct dendritic cell (DC) subsets, namely, plasmacytoid DCs (pDCs) and conventional DC subsets (cDC1s and cDC2s), is controlled by specific transcription factors. IRF8 is essential for the fate specification of cDC1s. However, how the expression of Irf8 is regulated is not fully understood. In this study, we identified TRIM33 as a critical regulator of DC differentiation and maintenance. TRIM33 deletion in Trim33fl/fl Cre-ERT2 mice significantly impaired DC differentiation from hematopoietic progenitors at different developmental stages. TRIM33 deficiency downregulated the expression of multiple genes associated with DC differentiation in these progenitors. TRIM33 promoted the transcription of Irf8 to facilitate the differentiation of cDC1s by maintaining adequate CDK9 and Ser2 phosphorylated RNA polymerase II (S2 Pol II) levels at Irf8 gene sites. Moreover, TRIM33 prevented the apoptosis of DCs and progenitors by directly suppressing the PU.1-mediated transcription of Bcl2l11, thereby maintaining DC homeostasis. Taken together, our findings identified TRIM33 as a novel and crucial regulator of DC differentiation and maintenance through the modulation of Irf8 and Bcl2l11 expression. The finding that TRIM33 functions as a critical regulator of both DC differentiation and survival provides potential benefits for devising DC-based immune interventions and therapies.

Dynamic atlas of immune cells reveals multiple functional features of macrophages associated with progression of pulmonary fibrosis.

Idiopathic pulmonary fibrosis (IPF) is a chronic interstitial lung disease with a high mortality rate and unclarified aetiology. Immune response is elaborately regulated during the progression of IPF, but immune cells subsets are complicated which has not been detailed described during IPF progression. Therefore, in the current study, we sought to investigate the role of immune regulation by elaborately characterize the heterogeneous of immune cells during the progression of IPF. To this end, we performed single-cell profiling of lung immune cells isolated from four stages of bleomycin-induced pulmonary fibrosis-a classical mouse model that mimics human IPF. The results revealed distinct components of immune cells in different phases of pulmonary fibrosis and close communication between macrophages and other immune cells along with pulmonary fibrosis progression. Enriched signals of SPP1, CCL5 and CXCL2 were found between macrophages and other immune cells. The more detailed definition of the subpopulations of macrophages defined alveolar macrophages (AMs) and monocyte-derived macrophages (mo-Macs)-the two major types of primary lung macrophages-exhibited the highest heterogeneity and dynamic changes in expression of profibrotic genes during disease progression. Our analysis suggested that Gpnmb and Trem2 were both upregulated in macrophages and may play important roles in pulmonary fibrosis progression. Additionally, the metabolic status of AMs and mo-Macs varied with disease progression. In line with the published data on human IPF, macrophages in the mouse model shared some features regarding gene expression and metabolic status with that of macrophages in IPF patients. Our study provides new insights into the pathological features of profibrotic macrophages in the lung that will facilitate the identification of new targets for disease intervention and treatment of IPF.

OX40L-Armed Oncolytic Virus Boosts T-cell Response and Remodels Tumor Microenvironment for Pancreatic Cancer Treatment.

Rationale: The resistance of pancreatic ductal adenocarcinoma (PDAC) to immunotherapies is caused by the immunosuppressive tumor microenvironment (TME) and dense extracellular matrix. Currently, the efficacy of an isolated strategy targeting stromal desmoplasia or immune cells has been met with limited success in the treatment of pancreatic cancer. Oncolytic virus (OV) therapy can remodel the TME and damage tumor cells either by directly killing them or by enhancing the anti-tumor immune response, which holds promise for the treatment of PDAC. This study aimed to investigate the therapeutic effect of OX40L-armed OV on PDAC and to elucidate the underlying mechanisms. Methods: Murine OX40L was inserted into herpes simplex virus-1 (HSV-1) to construct OV-mOX40L. Its expression and function were assessed using reporter cells, cytopathic effect, and immunogenic cell death assays. The efficacy of OV-mOX40L was then evaluated in a KPC syngeneic mouse model. Tumor-infiltrating immune and stromal cells were analyzed using flow cytometry and single-cell RNA sequencing to gain insight into the mechanisms of oncolytic virotherapy. Results: OV-mOX40L treatment delayed tumor growth in KPC tumor-bearing C57BL/6 mice. It also boosted the tumor-infiltrating CD4+ T cell response, mitigated cytotoxic T lymphocyte (CTL) exhaustion, and reduced the number of regulatory T cells. The treatment of OV-mOX40L reprogrammed macrophages and neutrophils to a more pro-inflammatory anti-tumor state. In addition, the number of myofibroblastic cancer-associated fibroblasts (CAF) was reduced after treatment. Based on single-cell sequencing analysis, OV-mOX40L, in combination with anti-IL6 and anti-PD-1, significantly extended the lifespan of PDAC mice. Conclusion: OV-mOX40L converted the immunosuppressive tumor immune microenvironment to a more activated state, remodeled the stromal matrix, and enhanced T cell response. OV-mOX40L significantly prolonged the survival of PDAC mice, either as a monotherapy or in combination with synergistic antibodies. Thus, this study provides a multimodal therapeutic strategy for pancreatic cancer treatment.

Protective effects of Fagopyrum dibotrys on oxidized oil-induced oxidative stress, intestinal barrier impairment, and altered cecal microbiota in broiler chickens.

The objective of this study was to evaluate protective effects of Fagopyrum dibotrys on antioxidant ability, intestinal barrier functions, and cecal microbiota in broiler chickens fed oxidized soybean oil. A total of 640 male Tiejiaoma broilers were randomly assigned to 8 treatments with 8 cages (10 birds per cage), as follows: birds fed basal diets containing fresh soybean oil and 0, 0.5, 1, or 2% F. dibotrys (FSCON, FSFAL, FSFAM, and FSFAH, respectively), and birds fed basal diets containing oxidized oil and 0, 0.5, 1, or 2% F. dibotrys (OSCON, OSFAL, OSFAM, and OSFAH). Oxidized oil significantly decreased transcription of Nrf2 and its downstream genes, including CAT and SOD1 in the jejunal mucosa, increased jejunal mucosa IL-6 mRNA expression, and decreased jejunal mucosa IL-22 mRNA expression and downregulated Claudin-1 and ZO-1; however, all these effects were reversed by F. dibotrys. Either 1 or 2% F. dibotrys alleviated the decreased liver SOD induced by oxidized oil on d 42. The decreased SOD and GPX, and increased MDA induced by oxidized oil were reversed by adding 1 or 2% F. dibotrys in jejunal mucosa. In addition, based on 16S rDNA, 2% F. dibotrys promoted the Firmicutes phylum and Candidatus_Arthromitus genera, but suppressed the Proteobacteria phylum and Streptococcus, Enterococcus, and Escherichia genera. In summary, oxidative stress induced by oxidized oil was ameliorated by F. dibotrys upregulating transcription of Nrf2 and its downstream genes to restore redox balance, reinforcing the intestinal barrier via higher expression of Claudin-1/ZO-1, ameliorating the inflammatory response by regulating expression of IL-6 and IL-22, and facilitating growth of Candidatus_arthromitus in the cecum. Therefore, F. dibotrys has potential as a feed additive for poultry by ameliorating oxidative stress caused by oxidized oil, enhancing barrier function, and improving gut microbiome composition.

Predictors of thromboembolic complications after stent-assisted coiling of acutely ruptured intracranial aneurysms: A retrospective multicenter study.

Background: Stent-assisted coiling (SAC) has been reported to safely and effectively treat wide-necked unruptured intracranial aneurysms. However, SAC of acutely ruptured aneurysms is controversial because of perioperative thromboembolic complications. We aimed to investigate the predictors of the thromboembolic complications after SAC of acutely ruptured aneurysms. Methods: We performed a retrospective multicenter analysis of 110 consecutive patients with ruptured intracranial aneurysms treated with SAC within 72 h of the onset of subarachnoid hemorrhage. Thromboembolic complications were defined as any angiographic filling defects at the aneurysms base or the distal artery during the stent treatment and the new onset of symptomatic ischemia and a new hypo-density in a vascular distribution confirmed by CT scan within 24 h of treatment. These patients were grouped into patients with thromboembolic complications and those without thromboembolic complications. A multivariate logistic regression analysis was performed to identify predictors of thromboembolic complications. Results: One hundred and one patients with 101 ruptured aneurysms were included in this study. 9 (8.9%) patients experienced thromboembolic complications. Patients with thromboembolic complications had a higher rate of unfavorable outcomes at discharge (P < 0.001) and at the last follow-up (p = 0.017). Of these patients, four patients presented with intraprocedural thrombus formation, and 5 experienced postprocedural ischemia. There was a trend toward thromboembolic complications in patients with a higher Fisher grade (p = 0.076) and those treated with intravenous tirofiban (p = 0.052). Patients with thromboembolic complications more often presented with poor grade clinical conditions (p = 0.005) and aneurysms with a large dome to neck ratio (p = 0.031). In the multivariate analysis, a worse World Federation World Federation of Neurological Societies (WFNS) grade (OR = 8.241; 95% CI 1.686-40.292; P = 0.009) and a larger dome to neck ratio (OR = 5.385; 95% CI 1.023-28.337; P = 0.047) were independent predictors of thromboembolic complications. Conclusion: Patients with thromboembolic complications are more likely to have an unfavorable outcome. A worse clinical condition before the treatment and a larger dome to neck ratio were independent predictors of thromboembolic complications after SAC of acutely ruptured intracranial aneurysms.

An integrated nanoflower-like MoS2@CuCo2O4 heterostructure for boosting electrochemical glucose sensing in beverage.

Excessive glucose in food poses a non-negligible threat to its inherent quality and human health, which makes it imperative to develop a highly sensitive sensor for real-time glucose detection. In this work, an integrated electrochemical glucose sensor based on a nanoflower-like MoS2@CuCo2O4 heterostructure was carefully constructed. Under optimal conditions, the as-fabricated sensor exhibited a high sensitivity of 1,303 µA mM-1 cm-2 over a wide range of 0.5-393.0 µmol/L, accompanied by a low determination limit (0.5 µmol/L) and short response time (2.1 s). The favorable sensing performance of the MoS2@CuCo2O4 nanocomposite-modified electrode in electrochemical analyses was attributed to the introduction of unique nanoflower-like heterostructure and the synergistic effects between MoS2 and CuCo2O4. Furthermore, the satisfactory applicability of this sensor in beverages was confirmed. These results demonstrate that the MoS2@CuCo2O4/GCE may be a promising platform for sensitive monitoring of glucose content in food samples.

An armed oncolytic virus enhances the efficacy of tumor-infiltrating lymphocyte therapy by converting tumors to artificial antigen-presenting cells in situ.

The full potential of tumor-infiltrating lymphocyte (TIL) therapy has been hampered by the inadequate activation and low persistence of TILs, as well as inefficient neoantigen presentation by tumors. We transformed tumor cells into artificial antigen-presenting cells (aAPCs) by infecting them with a herpes simplex virus 1 (HSV-1)-based oncolytic virus encoding OX40L and IL12 (OV-OX40L/IL12) to provide local signals for optimum T cell activation. The infected tumor cells displayed increased expression of antigen-presenting cell-related markers and induced enhanced T cell activation and killing in coculture with TILs. Combining OV-OX40L/IL12 and TIL therapy induced complete tumor regression in patient-derived xenograft and syngeneic mouse tumor models and elicited an antitumor immunological memory. In addition, the combination therapy produced aAPC properties in tumor cells, activated T cells, and reprogrammed macrophages to a more M1-like phenotype in the tumor microenvironment. This combination strategy unleashes the full potential of TIL therapy and warrants further evaluation in clinical studies.

Preventive Effects of Sesamol on Deep-Frying Oil-Induced Liver Metabolism Disorders by Altering Gut Microbiota and Protecting Gut Barrier Integrity.

SCOPE: The total polar components (TPC) in the edible oil are produced during the frying process, and excessive intake of TPC may lead to metabolic disorders. This study aims to investigate the preventive effects of sesamol, a functional component from sesame, on suppressing TPC production, and on the deep-frying oil (DFO)-induced liver lipid metabolism disorders and gut homeostasis disruption. METHODS AND RESULTS: Sesamol addition (0.2 mg mL-1 ) improves the quality of the soybean oil by reducing the production of TPC during the deep-frying process (180 °C for 40 h). The diet-induced obesity model is established by feeding mice with a high-fat diet for 8 weeks. Either sesamol pre-treated DFO or sesamol supplementation (0.2%, w/w) in the diet reduces the liver lipid accumulation in the obese mice by increasing lipolysis-related genes expression. Sesamol elevates the antioxidant enzyme activities, protectes the integrity of the jejunum and colon barrier, and enhances the relative abundance of Bifidobacterium and Akkermansia in obese mice. CONCLUSION: Sesamol suppresses TPC production and prolongs the use time of deep-frying oil. Meanwhile, sesamol can improve TPC-induced liver lipid metabolism disorders and gut dysbiosis, thus reducing the health risks associated with deep-fried food.

LRRK2 plays essential roles in maintaining lung homeostasis and preventing the development of pulmonary fibrosis.

Perturbation of lung homeostasis is frequently associated with progressive and fatal respiratory diseases, such as pulmonary fibrosis. Leucine-rich repeat kinase 2 (LRRK2) is highly expressed in healthy lungs, but its functions in lung homeostasis and diseases remain elusive. Herein, we showed that LRRK2 expression was clearly reduced in mammalian fibrotic lungs, and LRRK2-deficient mice exhibited aggravated bleomycin-induced pulmonary fibrosis. Furthermore, we demonstrated that in bleomycin-treated mice, LRRK2 expression was dramatically decreased in alveolar type II epithelial (AT2) cells, and its deficiency resulted in profound dysfunction of AT2 cells, characterized by impaired autophagy and accelerated cellular senescence. Additionally, LRRK2-deficient AT2 cells showed a higher capacity of recruiting profibrotic macrophages via the CCL2/CCR2 signaling, leading to extensive macrophage-associated profibrotic responses and progressive pulmonary fibrosis. Taken together, our study demonstrates that LRRK2 plays a crucial role in preventing AT2 cell dysfunction and orchestrating the innate immune responses to protect against pulmonary fibrosis.

High-throughput functional screening for next-generation cancer immunotherapy using droplet-based microfluidics.

Currently, high-throughput approaches are lacking in the isolation of antibodies with functional readouts beyond simple binding. This situation has impeded the next generation of cancer immunotherapeutics, such as bispecific T cell engager (BiTE) antibodies or agonist antibodies against costimulatory receptors, from reaching their full potential. Here, we developed a highly efficient droplet-based microfluidic platform combining a lentivirus transduction system that enables functional screening of millions of antibodies to identify potential hits with desired functionalities. To showcase the capacity of this system, functional antibodies for CD40 agonism with low frequency (<0.02%) were identified with two rounds of screening. Furthermore, the versatility of the system was demonstrated by combining an anti-Her2 × anti-CD3 BiTE antibody library with functional screening, which enabled efficient identification of active anti-Her2 × anti-CD3 BiTE antibodies. The platform could revolutionize next-generation cancer immunotherapy drug development and advance medical research.

Structural basis for SARS-CoV-2 neutralizing antibodies with novel binding epitopes.

The ongoing Coronavirus Disease 2019 (COVID-19) pandemic caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) threatens global public health and economy unprecedentedly, requiring accelerating development of prophylactic and therapeutic interventions. Molecular understanding of neutralizing antibodies (NAbs) would greatly help advance the development of monoclonal antibody (mAb) therapy, as well as the design of next generation recombinant vaccines. Here, we applied H2L2 transgenic mice encoding the human immunoglobulin variable regions, together with a state-of-the-art antibody discovery platform to immunize and isolate NAbs. From a large panel of isolated antibodies, 25 antibodies showed potent neutralizing activities at sub-nanomolar levels by engaging the spike receptor-binding domain (RBD). Importantly, one human NAb, termed PR1077, from the H2L2 platform and 2 humanized NAb, including PR953 and PR961, were further characterized and subjected for subsequent structural analysis. High-resolution X-ray crystallography structures unveiled novel epitopes on the receptor-binding motif (RBM) for PR1077 and PR953, which directly compete with human angiotensin-converting enzyme 2 (hACE2) for binding, and a novel non-blocking epitope on the neighboring site near RBM for PR961. Moreover, we further tested the antiviral efficiency of PR1077 in the Ad5-hACE2 transduction mouse model of COVID-19. A single injection provided potent protection against SARS-CoV-2 infection in either prophylactic or treatment groups. Taken together, these results shed light on the development of mAb-related therapeutic interventions for COVID-19.

A general Fc engineering platform for the next generation of antibody therapeutics.

Rationale: Fc engineering has become the focus of antibody drug development. The current mutagenesis and in silico protein design methods are confined by the limited throughput and high cost, while the high-throughput phage display and yeast display technologies are not suitable for screening glycosylated Fc variants. Here we developed a mammalian cell display-based Fc engineering platform. Methods: By using mammalian cell display and next generation sequencing, we screened millions of Fc variants for optimized affinity and specificity for FcγRIIIa or FcγRIIb. The identified Fc variants with improved binding to FcγRIIIa were substituted into trastuzumab and rituximab and the effector function of antibodies were examined in the PBMC-based assay. On the other hand, the identified Fc variants with selectively enhanced FcγRIIb binding were applied to CD40 agonist antibody and the activities of the antibodies were measured on different cell assays. The immunostimulatory activity of CD40 antibodies was also evaluated by OVA-specific CD8+ T cell response model in FcγR/CD40-humanized mice. Results: Using this approach, we screened millions of Fc variant and successfully identified several novel Fc variants with enhanced FcγRIIIa or FcγRIIb binding. These identified Fc variants displayed a dramatic increase in antibody-dependent cellular cytotoxicity in PBMC-based assay. Novel variants with selectively enhanced FcγRIIb binding were also identified. CD40 agonist antibodies substituted with these Fc variants displayed activity more potent than the parental antibody in the in vitro and in vivo models.Conclusions: This approach increased the throughput of Fc variant screening from thousands to millions magnitude, enabled screening variants containing multiple mutations and could be integrated with glycoengineering technology, represents an ideal platform for Fc engineering. The initial efforts demonstrated the capability of the platform and the novel Fc variants could be substituted into nearly any antibody for the next generation of antibody therapeutics.

An innovative evaluation method based on polymer mass detection to evaluate the contribution of microfibers from laundry process to municipal wastewater.

Clothes washing releases numerous microfibers, including microplastic fibers (MPFs). Although MPFs in laundry wastewater are an important source of microplastics (MPs) in wastewater treatment plants (WWTPs), credible quantitative assessments of their contributions remain limited. Polyester fiber is the most important textile fiber. Its component, polyethylene terephthalate (PET) polymer, can be quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The release of MPFs from polyester clothes through washing was quantified via simulation experiments, and the MPFs in two WWTPs were measured by microscopic counting and LC-MS/MS. Direct comparison of the abundances of PET MPFs in laundry wastewater and WWTP influents led to an undervalued contribution rate of 9%-11% of the PET MPFs in laundry wastewater to those in WWTP influents. However, comparison of the mass of PET MPFs in laundry wastewater and WWTPs influents revealed that the PET MPFs from laundry contributed approximately 50% of those in the WWTPs. The latter was confirmed by comparing the number of polyester fibers released during clothes washing to the calculated number of "model MPFs" in WWTPs according to the PET mass concentration. Based on the PET concentration, the annual discharge of PET MPs from WWTPs to the water environment could also be estimated.

Identification and immunogenic evaluation of T cell epitopes based on tembusu virus envelope protein in ducks.

Newly emerging tembusu virus (TMUV) is a severe threat to poultry industry and causes huge economic losses. Humoral and cell-mediated immunity are both play vital roles in TMUV infection. Up to now, there has been no report on identification of T cell epitopes of the TMUV. In this work, we identified T cell epitopes within TMUV envelope (E) protein using synthesized peptides predicted in silico. A total of ten peptides could stimulate TMUV-specific T cells in murine ELISPOT and duck lymphocyte proliferation assay. Subsequently, DNA vaccine containing these T cell epitopes was constructed (pVAX-T) and the expression of multiepitope protein was confirmed by transfection of BHK-21 cells in vitro. Ducks were administrated intramusclarly to evaluated the immunologic effect of pVAX-T. In ducks immunized with pVAX-T, antibody against TMUV was undetectable, but the expression level of cytokines (IL-2, IL-6, IFN-γ) was upregulated both in peripheral blood lymphocytes and spleen. Furthermore, TMUV challenge revealed that cell-mediated immune response sitmulated by pVAX-T contributed to protection against TMUV infection. The identification of these T cell epitopes will contribute to designing epitope vaccine for preventing infection of TMUV and possibly provide the basis for further studies on cell-mediate immune response activated by TMUV.

Screening and identification of B-cell epitopes within envelope protein of tembusu virus.

BACKGROUND: Tembusu virus is a newly emerging flavivirus that caused egg-drop syndrome in ducks in China. TMUV envelope protein is a major structural protein locates at the surface of tembusu virus particle. During tembusu virus infection, envelope protein plays a pivotal role in induction of neutralizing antibody. However, B cell epitopes within envelope protein have not been well studied. METHOD: A series of 13 peptides derived from E protein of tembusu virus were synthesized and screened by Dot blot with tembusu virus-positive duck serum. Potential B-cell epitopes were respectively fused with GST tag and expressed in E. coli. The immunogenicity and protective efficiency of epitopes were assessed in ducks. RESULTS: Dot blot assay identified the peptides P21 (amino acids 301-329), P23 (amino acids 369-387), P27 (amino acids 464-471) and P28 (amino acids 482-496) as potential B-cell epitopes within the envelope protein of tembusu virus. Immunization of prokaryotically expressed epitopes elicited specific antibodies in ducks and the specific antibody elicited by P21, P27 and P28 could neutralized tembusu virus. In addition, protective test suggested that P21 and P27 could completely protect immunized ducks from TMUV challenge. CONCLUSION: Four potential B cell epiotpes within tembusu virus envelope protein were identified and analyzed in vitro and in vivo. It was demonstrated that two of them (P21 and P27) could elicit neutralizing antibodies in ducks and offer complete protection against tembusu virus challenge. This findings will contribute to the development of epitope vaccine for tembusu virus prevention.

Elevated heat shock proteins in bipolar disorder patients with hypothalamic pituitary adrenal axis dysfunction.

BACKGROUND: Heat shock proteins (HSP) might be useful as biomarkers for bipolar disorder (BD) which would be clinically valuable since no reliable biomarker for BD has so far been identified. The purpose of this study was to assess the heat shock proteins CPN10, CPN60, and CPN70 as potential biomarkers of BD. METHODS: The study included 100 BD patients recruited from a hospital during 2012 and 2013. The study also included 94 healthy controls. Among the BD patients, 33 had abnormal hypothalamic-pituitary-adrenal (HPA) axis activity. Blood samples were obtained from the patients and controls. The chemiluminescence method, mass spectrometry, and flow cytometry were used for analysis. RESULTS: The BD patients compared with the controls had a significantly lower level of CPN10 and significantly higher levels of CPN60 and CPN70. The BD patients with abnormal HPA axis activity had a significantly lower level of CPN60 compared with the normal HPA axis activity group of BD patients. The CPN60 level significantly inversely correlated with adrenocorticotropic hormone (ACTH) level in patients with bipolar depression and in patients with bipolar hypomania, and CPN70 significantly correlated with ACTH level in patients with bipolar depression and hypomania. CONCLUSIONS: Our findings suggest that the heat shock proteins CPN10, CPN60, and CPN70 might have potential as biomarkers for BD and CPN60 blood level might distinguish patients with abnormal HPA axis activity from those with normal HPA axis activity.

Identification of Glucose-Regulated Protein 78 (GRP78) as a Receptor in BHK-21 Cells for Duck Tembusu Virus Infection.

Since 2010, outbreak and spread of tembusu virus (TMUV) caused huge losses to the breeding industry of waterfowl in several provinces of China. In this study, we identify the glucose-regulated protein 78 (GRP78) as a receptor in BHK-21 cells for duck TMUV infection. Using cell membrane from BHK-21 cells, a TMUV-binding protein of approximately 70 kDa was observed by viral overlay protein binding assay (VOPBA). LC-MS/MS analysis and co-immunoprecipitation identified GRP78 as a protein interacting with TMUV. Antibody against GRP78 inhibited the binding of TMUV to the cell surface of BHK-21 cells. Indirect immunofluorescence studies showed the colocalization of GRP78 with TMUV in virus-infected BHK-21 cells. We found that GRP78 over-expression increased TMUV infection, whereas GRP78 knockdown by using a specific small interfering RNA inhibited TMUV infection in BHK-21 cells. Taken together, our results indicate that GRP78 is a novel host factor involved in TMUV entry.

The relationship between brain reaction and English reading tests for non-native English speakers.

This research analyzed the brain activity of non-native English speakers while engaged in English reading tests. The brain wave event-related potentials (ERPs) of participants were used to analyze the difference between making correct and incorrect choices on English reading test items. Three English reading tests of differing levels were designed and 20 participants, 10 males and 10 females whose ages ranged from 20 to 24, voluntarily participated in the experiment. Experimental results were analyzed by performing independent t-tests on the ERPs of participants for gender, difficulty level, and correct versus wrong options. Participants who chose incorrect options elicited a larger N600, verifying results found in the literature. Another interesting result was found: For incorrectly answered items, different areas of brain showing a significant difference in ERPs between the chosen and non-chosen options corresponded to gender differences; for males, this area was located in the right hemisphere whereas for females, it was located in the left. Experimental results imply that non-native English speaking males and females employ different areas of the brain to comprehend the meaning of difficult items.