Intravenous Immunoglobulin Gamma (IVIG) versus IVIG Plus Infliximab in Young Children with Kawasaki Disease.

BACKGROUND Kawasaki disease (KD) is a serious disease characterized by systemic lesions of the skin and mucous membranes, as well as lymphomas and vascular inflammation. KD threatens the health and lives of children, especially young ones. Here, we compared the therapeutic effects of single intravenous immunoglobulin gamma (IVIG) vs. a combination of IVIG and infliximab in young children with Kawasaki disease (KD). MATERIAL AND METHODS A total of 154 children with KD, younger than 5 years old, were enrolled in the study from January 2013 to January 2017. The patients were randomly divided into an IVIG group and a combination of IVIG and infliximab treatment group. After systematic treatments, the therapeutic indicators of the 2 groups were compared. During the treatment process, body temperature and other important inflammatory indicators, including C-reactive protein (CRP), white blood cell count (WBC), and tumor necrosis factor alpha (TNF-α), were monitored in the first 4 days. RESULTS There were fewer refractory KD patients in the combined treatment group than in the IVIG group (4 vs. 14, p<0.001). KD patients in the combined treatment group had better outcomes with shorter fever durations and hospital stays, as well as less coronary artery dilation. However, there was no obvious differences in the incidence rate of coronary artery aneurysms between the 2 groups (p>0.05). Costs of administration were similar between groups (p>0.05). Body temperature, CRP, WBC, and TNF-α in the combined therapy group all showed an earlier drop than in the IVIG group, indicating a more effective anti-inflammation effect. CONCLUSIONS The introduction of IVIG combined with infliximab in the treatment of young children with KD has more advantages than single IVIG therapy and can be considered as a preferred treatment for KD. However, it would be necessary to further investigate whether there is a significant difference in aneurysm frequency and long-term outcome between these 2 strategies among a larger number of patients.

Rapid development of double-hit mRNA antibody cocktail against orthopoxviruses.

The Orthopoxvirus genus, especially variola virus (VARV), monkeypox virus (MPXV), remains a significant public health threat worldwide. The development of therapeutic antibodies against orthopoxviruses is largely hampered by the high cost of antibody engineering and manufacturing processes. mRNA-encoded antibodies have emerged as a powerful and universal platform for rapid antibody production. Herein, by using the established lipid nanoparticle (LNP)-encapsulated mRNA platform, we constructed four mRNA combinations that encode monoclonal antibodies with broad neutralization activities against orthopoxviruses. In vivo characterization demonstrated that a single intravenous injection of each LNP-encapsulated mRNA antibody in mice resulted in the rapid production of neutralizing antibodies. More importantly, mRNA antibody treatments showed significant protection from weight loss and mortality in the vaccinia virus (VACV) lethal challenge mouse model, and a unique mRNA antibody cocktail, Mix2a, exhibited superior in vivo protection by targeting both intracellular mature virus (IMV)-form and extracellular enveloped virus (EEV)-form viruses. In summary, our results demonstrate the proof-of-concept production of orthopoxvirus antibodies via the LNP-mRNA platform, highlighting the great potential of tailored mRNA antibody combinations as a universal strategy to combat orthopoxvirus as well as other emerging viruses.

Multiple underlying images tuned by Mn-doped Zn-Cu-In-S quantum dots.

In this study, ZnS capped Cu-In-S (ZCIS) quantum dots doped with Mn ions are synthesized by a thermal injection method, with luminescence covering almost the entire visible area. The large Stokes shift effectively inhibits the self-absorption effect under luminescence, and the quantum yield of ZCIS quantum dots increased from 38% to 50% after ZnS capping and further to 69% after doping with Mn. First, red-, yellow-, and blue-emitting quantum dots were synthesized and then, polychromatic ensembles were obtained by mixing the trichromatic quantum dots in a different ratio. Using the home-built inkjet printer, multilayered and multicolor mixed patterns were obtained for information pattern storage and multilayer pattern recognition and reading.

CD32 Expression by CD4+ T and CD8+ T Lymphocytes Is Increased in Patients with Chronic Hepatitis B Virus Infection.

FcγR is expressed by many immune cells and plays an important role in the immune response to hepatitis B virus (HBV) infection. CD32 belongs to the FcγR family. This study aimed to observe changes in CD32 expression by CD4+ T and CD8+ T lymphocytes in chronic HBV infection patients and evaluate the clinical utility of CD4+ T and CD8+ T CD32 expression to assess the severity of liver injury in chronic HBV-infected patients. A total of 68 chronic HBV patients and 40 healthy individuals were recruited, and the median fluorescence intensity (MFI) of CD32 expression on CD4+ T, CD8+ T lymphocytes was measured using flow cytometry and the CD4+ T, CD8+ T CD32 index was calculated. The reactivity of the healthy individual lymphocytes to mixed patients' plasma containing HBV was observed. Finally, the correlation between CD4+ T, CD8+ T lymphocytes CD32 MFI and liver function indicator levels was analyzed. The CD4+ T, CD8+ T CD32 MFI and index were significantly elevated in HBV patient groups than in normal control group (p < 0.001, for all). Furthermore, the CD32 MFI of healthy persons' CD4+ T and CD8+ T lymphocytes were remarkably increased when stimulated with mixed patients' plasma containing high HBV copies (p < 0.001; P < 0.001). More importantly, in HBV patients, there was a significant positive correlation between CD4+ T, CD8+ T CD32 MFI and the level of serum aspartate aminotransferase (p < 0.05, p < 0.05). In conclusion, the increased expression of CD32 on CD4+ T and CD8+ T lymphocytes might be potential promising biomarkers for the severity of liver function impairment in chronic HBV patients.

PA-E18G substitution in influenza A virus confers resistance to ZX-7101, a cap-dependent endonuclease inhibitor.

Cap-dependent endonuclease (CEN) in the polymerase acidic protein (PA) of influenza A virus (IAV) represents a promising drug target due to its critical role in viral gene transcription. The CEN inhibitor, baloxavir marboxil (BXM), was approved in Japan and the US in 2018 and several other countries subsequently. Along with the clinical use of BXM, the emergence and spread of IAV variants with reduced susceptibility to BXM have aroused serious concern. Herein, we comprehensively characterized the in vitro and in vivo antiviral activities of ZX-7101A, an analogue of BXM. The active form of prodrug ZX-7101 showed broad-spectrum antiviral potency against various IAV subtypes, including pH1N1, H3N2, H7N9 and H9N2, in MDCK cells, and the 50% effective concentration (EC50) was calculated to nanomole level and comparable to that of baloxavir acid (BXA), the active form of BXM. Furthermore, in vivo assays showed that administration of ZX-7101A conferred significant protection against lethal pH1N1 challenge in mice, with reduced viral RNA loads and alleviated pulmonary damage. Importantly, serial passaging of H1N1 virus in MDCK cells under selection pressure of ZX-7101 led to a resistant variant at the 15th passage. Reverse genetic and sequencing analysis demonstrated that a single E18G substitution in the PA subunit contributed to the reduced susceptibility to both ZX-7101 and BXA. Taken together, our results not only characterized a new CEN inhibitor of IAV but also identified a novel amino acid substitution responsible for CEN inhibitor resistance, which provides critical clues for future drug development and drug resistance surveillance.

Rational development of multicomponent mRNA vaccine candidates against mpox.

The re-emerging mpox (formerly monkeypox) virus (MPXV), a member of Orthopoxvirus genus together with variola virus (VARV) and vaccinia virus (VACV), has led to public health emergency of international concern since July 2022. Inspired by the unprecedent success of coronavirus disease 2019 (COVID-19) mRNA vaccines, the development of a safe and effective mRNA vaccine against MPXV is of high priority. Based on our established lipid nanoparticle (LNP)-encapsulated mRNA vaccine platform, we rationally constructed and prepared a panel of multicomponent MPXV vaccine candidates encoding different combinations of viral antigens including M1R, E8L, A29L, A35R, and B6R. In vitro and in vivo characterization demonstrated that two immunizations of all mRNA vaccine candidates elicit a robust antibody response as well as antigen-specific Th1-biased cellular response in mice. Importantly, the penta- and tetra-component vaccine candidates AR-MPXV5 and AR-MPXV4a showed superior capability of inducing neutralizing antibodies as well as of protecting from VACV challenge in mice. Our study provides critical insights to understand the protection mechanism of MPXV infection and direct evidence supporting further clinical development of these multicomponent mRNA vaccine candidates.

Identification of Six N7-Methylguanosine-Related miRNA Signatures to Predict the Overall Survival and Immune Landscape of Triple-Negative Breast Cancer through In Silico Analysis.

Triple-negative breast cancer (TNBC) is a widely prevalent breast cancer, with a mortality rate of up to 25%. TNBC has a lower survival rate, and the significance of N7-methylguanosine (m7G) modification in TNBC remains unclear. Thus, this study is aimed at investigating m7G-related miRNAs in TNBC patients through in silico analysis. In our research, RNA sequencing and clinical data were obtained from The Cancer Genome Atlas (TCGA) database. The miRNAs targeting typical m7G modification regulators Methyltransferase-like 1 (METTL1) and WD repeat domain 4 (WDR4) were predicted on the TargetScan website. A miRNA risk model was built, and its prognostic value was evaluated by R soft packages. Single-sample gene set enrichment analysis was used to assess immune infiltration, and further expression of immune checkpoints was investigated. As a result, miR-421, miR-5001-3p, miR-4326, miR-1915-3p, miR-3177-5p, and miR-4505 were identified to create the risk model. A nomogram consisting of the stage N and risk model predicted overall survival effectively among TNBC patients. Treg and TIL were shown to be strongly linked to the risk model, and the high-risk group had higher levels of four immune checkpoints expression (CD28, CTLA-4, ICOS, and TNFRSF9). A risk model consisting of m7G-related miRNAs was constructed. The findings of the current study could be used as a prognostic biomarker and can provide a novel immunotherapy insight for TNBC patients.

Integrated analysis of fibroblasts molecular features in papillary thyroid cancer combining single-cell and bulk RNA sequencing technology.

Background: Papillary thyroid cancer (PTC) is the most common pathological type of thyroid cancer with a high incidence globally. Increasing evidence reported that fibroblasts infiltration in cancer was correlated with prognostic outcomes. However, fibroblasts related study in thyroid cancer remains deficient. Methods: Single-cell sequencing data of PTC were analyzed by Seurat R package to explore the ecosystem in PTC and identify fibroblasts cluster. The expression profiles and prognostic values of fibroblast related genes were assessed in TCGA dataset. A fibrosis score model was established for prognosis prediction in thyroid cancer patients. Differentially expressed genes and functional enrichment between high and low fibrosis score groups in TCGA dataset were screened. The correlation of immune cells infiltration and fibrosis score in thyroid cancer patients was explored. Expression levels and prognostic values of key fibroblast related factor were validated in clinical tissues another PTC cohort. Results: Fibroblasts were highly infiltrated in PTC and could interact with other type of cells by single-cell data analysis. 34 fibroblast related terms were differentially expressed in thyroid tumor tissues. COX regression analysis suggested that the constructed fibrosis score model was an independent prognostic predictor for thyroid cancer patients (HR = 5.17, 95%CI 2.31-11.56, P = 6.36E-05). Patients with low fibrosis scores were associated with a significantly better overall survival (OS) than those with high fibrosis scores in TCGA dataset (P = 7.659E-04). Specific immune cells infiltration levels were positively correlated with fibrosis score, including monocytes, M1 macrophages and eosinophils. Conclusion: Our research demonstrated a comprehensive horizon of fibroblasts features in thyroid cancer microenvironment, which may provide potential value for thyroid cancer treatment.

High-Luminescence Electrospun Polymeric Microfibers In Situ Embedded with CdSe Quantum Dots with Excellent Environmental Stability for Heat and Humidity Wearable Sensors.

In this paper, hydrophobic luminescent CdSe quantum dots are successfully dispersed in a mixture of styrene and methyl methacrylate through the oleic to methacrylic acid ligand exchange. Further in situ solution polymerization of the quantum dots in a mixture of styrene and methyl methacrylate followed by electrospinning allowed us to prepare luminescence hybrid styrene-co-methyl methacrylate fibers embedded with quantum dots. CdSe@P(S+MMA) hybrid fibers with 27% quantum yield showed excellent moisture, heat and salt resistance with a photoluminescence output below 120 °C. When dry heated, the hybrid fibers of the fluorescence signals decreased with temperature to 79%, 40%, 28%, 20% and 13% at 120 °C, 140 °C, 160 °C, 180 °C and 200 °C, respectively, due the to the chemical degradation of CdSe QDs. Such hybrid fibers show the potential to manufacture wearable moisture- and heat-sensing protective clothing in a 120-200 °C range due to the thermal-induced quenching of quantum dot photoluminescence.

Analysis of the Gut Microbiota and Inflammatory Factors in mGluR5-Knockout Mice.

INTRODUCTION: Accumulating evidence indicates that the glutamatergic system plays an important role in the development of depression. Notably, the antidepressant effect of metabotropic glutamate receptor 5 (mGluR5) modulation is inconsistent across studies. Here, we attempted to identify the involvement of the gut microbiota and inflammation in mGluR5-/- mice. METHODS: mGluR5-/- mice and their wild-type littermates were used in our study. We used the open field (OF) and elevated plus maze (EPM) tests to assess anxiety-like behaviors, and we used the two-day forced swim test (FST) and tail suspension test (TST) to test despair-like behaviors. 16S rDNA was used to analyze the gut microbiota. Enzyme-linked immunosorbent assays (ELISAs) were used to measure the levels of inflammatory factors. Western blotting was used to detect the levels of various proteins. RESULTS: mGluR5-/- mice had no significant increase or decrease of despair-like behavior in the absence of stress exposure. However, mGluR5-/- mice exhibited despair-like behaviors following stress exposure. No significant changes in other glutamate receptors or representative synaptic proteins were detected in the prefrontal cortex (PFC) or hippocampus of mGluR5-/- mice. Very similar bacterial groups were observed in mGluR5-/- mice and wild-type controls. In addition, there was no significant difference in the α-diversity of the microbiota between mGluR5-/- mice and wild-type controls. The levels of all measured cytokines (IL-1ß, IL-2, IL-4, IL-6, IL-10, and TNF-α) did not change significantly in the PFCs or colons of mGluR5-/- mice. CONCLUSION: In conclusion, we deduced that mGluR5-/- mice are susceptible to despair-like behavior. The systemic knockout of mGluR5 did not affect the gut microbiota or inflammatory factors in mice.

Effect of a low-voltage electrical stimulation on yak meat tenderness during postmortem aging.

This study evaluates the effect of a low-voltage electrical stimulation (ES) on the tenderness of yak longissimus muscle (LM). Samples from 16 yak bulls were divided into four treatment groups: normal chilling (NC), ES and chilling (ES & C) for 72 s (ES &C 72 s), ES & C for 90 s (ES & C 90 s), and ES & C for 108 s (ES & C 108 s). The temperature, the pH, the glycogen content, the Warner-Bratzler shear force (WBSF), the myofibril fragmentation index (MFI), and the muscle ultrastructure were determined during the course of postmortem aging. ES caused a rapid decrease in the pH to form a high-temperature and low-pH environment. The glycogen content gradually decreased with aging. The WBSF value of the ES & C groups was significantly lower than for the NC group (p < .05). The MFI values of ES & C groups after 24 hr postmortem aging were significantly higher than for the NC group. We concluded that ES improved yak meat tenderness during postmortem aging and that the different duration time by ES indicated different effects, and its affect was remarkable in the ES & C 90 s.

Brain mGluR5 in Shank3B-/- Mice Studied With in vivo [18F]FPEB PET Imaging and ex vivo Immunoblotting.

Although several studies have found that metabotropic glutamate 5 receptor (mGluR5) may play an important role in autism spectrum disorders (ASD), the mechanisms remain unclear. Here, we used a Shank3 gene complete knockout mouse model (Shank3B-/-) to explore the change in mGluR5 in the brain. To assess whether deletion of Shank3 in mice results in ASD-like behavior, we conducted a battery of behavioral experiments to characterize Shank3B-/- mice, including repetitive grooming behavior tests, three-chamber tests and resident-intruder tests. Wild-type C57/BL6 and Shank3B-/- mice underwent PET scans with [18F]FPEB, which was highly specific to mGluR5. Mouse brains were extracted post-scan, and mGluR5 protein levels were assessed by immunoblotting. The binding potential (BPnd) of mGluR5 was rich in the hippocampus, thalamus, striatum, and amygdala. More importantly, Shank3B-/- mice showed significantly increased BPnd compared to the control mice in these brain regions. Immunoblotting revealed elevated mGluR5 levels in the hippocampus, thalamus, and amygdala but not in the striatum compared with control mice. These findings indicated that [18F]FPEB could visualize mGluR5 in the mouse brain. The deficiency of Shank3 can impair mGluR5 expression in multiple brain regions. Future work is also needed to understand the reasons for different results between in vivo PET and ex vivo immunoblotting.

Study on the effect of reactive oxygen species-mediated oxidative stress on the activation of mitochondrial apoptosis and the tenderness of yak meat.

This study investigated the effect of reactive oxygen species-mediated oxidative stress on activation of mitochondrial apoptosis and tenderness of yak meat during postmortem ageing. Oxidative stress degree, Ca2+ levels, membrane permeability transition pore opening, mitochondrial membrane potential, apoptotic factors and the shear force were examined. Results showed that the ROS generated by H2O2 significantly increased mitochondrial oxidative stress by decreasing the activities of superoxide dismutase, catalase and glutathione peroxidase, and increasing lipid peroxidation. Furthermore, oxidative stress enhanced Ca2+ production and cytochrome c release, changed the levels of Bcl-2 family proteins and activated caspase-9 and -3 activities. Ultimately, oxidative stress increased the apoptosis rate and tenderness of yak meat. These observations confirmed that ROS-mediated oxidative stress participates in the activation of the apoptotic cascade reaction involving Ca2+ and Bcl-2 family proteins. The results further suggested that ROS-mediated oxidative stress plays a significant role in meat tenderization through the mitochondrial apoptotic pathway.

Effect of mitochondrial apoptotic activation through the mitochondrial membrane permeability transition pore on yak meat tenderness during postmortem aging.

The effect of membrane permeability transition pore dependent mitochondrial apoptotic activation on yak meat tenderness was investigated. Results indicate that MPTP opening increased significantly and the mitochondrial membrane potential decreased markedly in the early aging process (P<0.05). Cytochrome c was released from the mitochondria to the cytoplasm via the MPTP in the early period. Meanwhile, the activation of procaspase-9 occurred earlier than that of procaspase-3. Cyclosporin A suppressed the MPTP opening, depolarization of the mitochondrial membrane potential, activities of caspase-9 and caspase-3, apoptosis rate, myofibril fragmentation index, reactive oxygen species generation, and Ca2+ levels. These results demonstrated that MPTP mediated the release of cytochrome c in the mitochondrial apoptotic pathway. Furthermore, yak meat tenderness was improved by mitochondrial apoptotic pathway during aging. MPTP opening may be influenced by the ROS generation and Ca2+ overloading in yak meat during postmortem aging.