Insight into selectivity of photocatalytic methane oxidation to formaldehyde on tungsten trioxide.

Tungsten trioxide (WO3) has been recognized as the most promising photocatalyst for highly selective oxidation of methane (CH4) to formaldehyde (HCHO), but the origin of catalytic activity and the reaction manner remain controversial. Here, we take {001} and {110} facets dominated WO3 as the model photocatalysts. Distinctly, {001} facet can readily achieve 100% selectivity of HCHO via the active site mechanism whereas {110} facet hardly guarantees a high selectivity of HCHO along with many intermediate products via the radical way. In situ diffuse reflectance infrared Fourier transform spectroscopy, electron paramagnetic resonance and theoretical calculations confirm that the competitive chemical adsorption between CH4 and H2O and the different CH4 activation routes on WO3 surface are responsible for diverse CH4 oxidation pathways. The microscopic mechanism elucidation provides the guidance for designing high performance photocatalysts for selective CH4 oxidation.

Coronin 2B deficiency induces nucleolar stress and neuronal apoptosis.

In eukaryotes, the nucleolus is the critical non-membranous organelle within nuclei that is responsible for ribosomal DNA (rDNA) transcription and ribosome biogenesis. The transcription of rDNA, a rate-limiting step for ribosome biogenesis, is tightly regulated to meet the demand for global protein synthesis in response to cell physiology, especially in neurons, which undergo rapid changes in morphology and protein composition during development and synaptic plasticity. However, it is unknown how the pre-initiation complex for rDNA transcription is efficiently assembled within the nucleolus in neurons. Here, we report that the nucleolar protein, coronin 2B, regulates rDNA transcription and maintains nucleolar function through direct interaction with upstream binding factor (UBF), an activator of RNA polymerase I transcriptional machinery. We show that coronin 2B knockdown impairs the formation of the transcription initiation complex, inhibits rDNA transcription, destroys nucleolar integrity, and ultimately induces nucleolar stress. In turn, coronin 2B-mediated nucleolar stress leads to p53 stabilization and activation, eventually resulting in neuronal apoptosis. Thus, we identified that coronin 2B coordinates with UBF to regulate rDNA transcription and maintain proper nucleolar function in neurons.

A high-permeability method for extracting purple yam saponins based on ultrasonic-assisted natural deep eutectic solvent.

The extraction of active ingredients from traditional Chinese medicine has received considerable attentions. In this study, 16 kinds of natural deep eutectic solvent (NADES) with ultrasonic were selected to extract saponins from purple yam root and the extraction mechanism was investigated. The results showed that chloride/acrylic acid (1:2; n/n) had the highest extraction yield for saponins. The optimal extraction process parameters were 24% water content, 20 mL/g liquid-solid ratio, and ultrasonic extraction for 85 min (81 °C, 600 W). The extraction rate (ER) of purple yam saponins was 0.935%, close to the fitted result of 96.5 mg/g. Molecular dynamics simulations and FT-IR results showed that the NADES may extract the saponin constituents from purple yam through hydrogen bonding. Compared with traditional extraction methods and molecularly imprinted polymer methods, NADES has a higher ER and lower cost (1.53 $/g), which provides a reference for subsequent industrial quantitative production.

Machine Learning Radiomics-Based Prediction of Non-sentinel Lymph Node Metastasis in Chinese Breast Cancer Patients with 1-2 Positive Sentinel Lymph Nodes: A Multicenter Study.

RATIONALE AND OBJECTIVES: This study aimed to construct a machine learning radiomics-based model using dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) images to evaluate non-sentinel lymph node (NSLN) metastasis in Chinese breast cancer (BC) patients who underwent total mastectomy (TM) and had 1-2 positive sentinel lymph nodes (SLNs). MATERIALS AND METHODS: In total, 494 patients were retrospectively enrolled from two hospitals, and were divided into the training (n = 286), internal validation (n = 122), and external validation (n = 86) cohorts. Features were extracted from DCE-MRI images for each patient and screened. Six ML classifies were trained and the best classifier was evaluated to calculate radiomics (Rad)-scores. A combined model was developed based on Rad-scores and clinical risk factors, then the calibration, discrimination, reclassification, and clinical usefulness were evaluated. RESULTS: 14 radiomics features were ultimately selected. The random forest (RF) classifier showed the best performance, with the highest average area under the curve (AUC) of 0.833 in the validation cohorts. The combined model incorporating RF-based Rad-scores, tumor size, lymphovascular invasion, and proportion of positive SLNs resulted in the best discrimination ability, with AUCs of 0.903, 0.890, and 0.836 in the training, internal validation, and external validation cohorts, respectively. Furthermore, the combined model significantly improved the classification accuracy and clinical benefit for NSLN metastasis prediction. CONCLUSION: A RF-based combined model using DCE-MRI images exhibited a promising performance for predicting NSLN metastasis in Chinese BC patients who underwent TM and had 1-2 positive SLNs, thereby aiding in individualized clinical treatment decisions.

Amlodipine inhibits the proliferation and migration of esophageal carcinoma cells through the induction of endoplasmic reticulum stress.

BACKGROUND: L-type calcium channels are the only protein channels sensitive to calcium channel blockers, and are expressed in various cancer types. The Cancer Genome Atlas database shows that the mRNA levels of multiple L-type calcium channel subunits in esophageal squamous cell carcinoma tumor tissue are significantly higher than those in normal esophageal epithelial tissue. Therefore, we hypothesized that amlodipine, a long-acting dihydropyridine L-type calcium channel blocker, may inhibit the occurrence and development of esophageal cancer (EC). AIM: To investigate the inhibitory effects of amlodipine on EC through endoplasmic reticulum (ER) stress. METHODS: Cav1.3 protein expression levels in 50 pairs of EC tissues and corresponding paracancerous tissues were examined. Subsequently, the inhibitory effects of amlodipine on proliferation and migration of EC cells in vitro were detected using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide and Transwell assays. In vivo experiments were performed using murine xenograft model. To elucidate the underlying mechanisms, in vitro cell studies were performed to confirm that ER stress plays a role in inhibition proliferation and migration of EC cells treated with amlodipine. RESULTS: The expression level of Cav1.3 in esophageal carcinoma was 1.6 times higher than that in paracancerous tissues. Amlodipine treatment decreased the viability of esophageal carcinoma cells in a dose- and time-dependent manner. In vivo animal experiments also clearly indicated that amlodipine inhibited the growth of EC tumors in mice. Additionally, amlodipine reduces the migration of tumor cells by inhibiting epithelial-mesenchymal transition (EMT). Mechanistic studies have demonstrated that amlodipine induces ER stress-mediated apoptosis and suppresses EMT. Moreover, amlodipine-induced autophagy was characterized by an increase in autophagy lysosomes and the accumulation of light chain 3B protein. The combination of amlodipine with the ER stress inhibitor 4-phenylbutyric acid further confirmed the role of the ER stress response in amlodipine-induced apoptosis, EMT, and autophagy. Furthermore, blocking autophagy increases the ratio of apoptosis and migration. CONCLUSION: Collectively, we demonstrate for the first time that amlodipine promotes apoptosis, induces autophagy, and inhibits migration through ER stress, thereby exerting anti-tumor effects in EC.

MiR-4521 affects the propagation of Cryptosporidium parvum in HCT-8 cells through targeting foxm1 by regulating cell apoptosis.

Cryptosporidium parvum could regulate the expression of microRNAs of epithelial cells to facilitate its intracellular propagation. MiR-4521 has been reported to play an important role during the development and progression of tumors and infectious diseases by regulating cell proliferation, apoptosis, and autophagy. However, the implication of miR-4521 during C. parvum infection was still unknown. In this study, the expression of miR-4521 was found to be upregulated in HCT-8 cells infected with C. parvum from 8 h post-infection (pi) to 48 hpi, and its upregulation would be related with the TLR/NF-κB signal pathway during C. parvum infection. One potential target of miR-4521, foxm1, was down-regulated in HCT-8 cells from 24 hpi to 48 hpi, and the expression of foxm1 was negatively regulated by miR-4521. The target relationship between miR-4521 and foxm1 was further validated by using dual luciferase reporter assay. Further studies showed that miR-4521 promoted the propagation of C. parvum in HCT-8 cells through targeting foxm1 by regulating BCL2-mediating cell apoptosis. These results contribute to further understanding of the regulatory mechanisms of host miRNAs during Cryptosporidium infection.

Golden-angle radial sparse parallel magnetic resonance imaging of rectal perfusion: utility in the diagnosis of poorly differentiated rectal cancer.

Background: The objective of this retrospective investigation is to evaluate the diagnostic efficacy of a dual-parameter strategy that integrates either time-resolved angiography with stochastic trajectories (TWIST) or golden-angle radial sparse parallel (GRASP)-derived dynamic contrast agent-enhanced magnetic resonance imaging (DCE-MRI) with diffusion-weighted imaging (DWI) for the identification of poorly differentiated rectal cancer (RC). The purpose of this investigation is to contrast the aforementioned methodology with conventional single-factor assessments that rely solely on DWI, and ascertain its comparative efficacy. Methods: This study was not registered on a clinical trial platform. Consecutive individuals diagnosed with non-mucinous rectal adenocarcinoma through endoscopy-guided biopsy between December 2020 and October 2022 were involved in our study. These patients had also undergone DCE-MRI and DWI. The perfusion metrics of influx forward volume transfer constant (Ktrans) and rate constant (Kep), along with the apparent diffusion coefficient (ADC), were quantified by a pair of investigators. The study compared the area under the curve (AUC) of the receiver operating characteristic (ROC) for both sequences to identify poorly differentiated RC. The investigation incorporated patients who fulfilled the specified criteria. The inclusion criteria for the investigation were as follows: (I) a diagnosis of RC proved through pathological examination, either via endoscopically-guided biopsy or surgical resection; (II) availability of complete MRI images; (III) absence of any prior history of neoadjuvant chemoradiotherapy during the MRI scan. Results: Our investigation comprised a total of 179 participants. Compared to diffusion parameter alone, an integrated assessment of diffusion parameter (ADC) and perfusion parameters (Ktrans or Kep) obtained with GRASP leads to a superior diagnostic accuracy (AUC, 0.97±0.02 vs. 0.89±0.03, 0.97±0.02 vs. 0.89±0.03, P=0.005 and 0.003, respectively); however, there was no additional benefit from ADC with perfusion parameters obtained from TWIST (Ktrans or Kep) (AUC, 0.93±0.04 vs. 0.89±0.03, 0.93±0.03 vs. 0.89±0.03; P= 0.955 and 0.981, respectively, for the integration of ADC with Ktrans and Kep). Conclusions: By integrating diffusion and perfusion features into a dual-parameter model, the GRASP method enhances the diagnostic efficacy of MRI in discriminating RCs with poor differentiation. Conversely, the TWIST approach did not yield the aforementioned outcome.

Predicting lymphovascular invasion in rectal cancer: evaluating the performance of golden-angle radial sparse parallel MRI for rectal perfusion assessment.

This study aims to determine whether the dual-parameter approach combined with either time-resolved angiography with stochastic trajectories (TWIST) or golden-angle radial sparse parallel (GRASP) and diffusion-weighted imaging (DWI) has superior diagnostic performance in predicting pathological lymphovascular invasion (pLVI) rectal cancer when compared with traditional single-parameter evaluations using DWI alone. Patients with pathologically confirmed rectal cancer were enrolled. Perfusion (influx forward volume transfer constant [Ktrans] and rate constant [Kep]) and apparent diffusion coefficient (ADC) were measured by two researchers. For both sequences, areas under receiver operating characteristic (ROCs) to predict pLVI-positive rectal cancer were compared. A total of 179 patients were enrolled in our study. A combined analysis of ADC and perfusion parameters (Ktrans) acquired with GRASP yielded a higher diagnostic performance compared with diffusion parameters alone (area under the curve, 0.91 ± 0.03 vs. 0.71 ± 0.06, P < 0.001); However, ADC with GRASP-acquired Kep and ADC with TWIST-acquired perfusion parameters (Ktrans or Kep) did not offer any additional benefit. The Ktrans of the GRASP technique improved the diagnostic performance of multiparametric MRI to predict rectal cancers with pLVI-positive. In contrast, TWIST did not achieve this effect.

Design of diversified chimeric antigen receptors through rational module recombination.

Chimeric antigen receptor (CAR)-T cells have shown great promise in cancer therapy. However, the anti-tumor efficiency is limited due to the CAR-induced T cell apoptosis or exhaustion. The intracellular domain of CAR comprised of various signaling modules orchestrates CAR-T cell behaviors. The modularity of CAR signaling domain functions as the "mainboard" to assemble diversified downstream signaling components. Here, we implemented the modular recombination strategy to construct a library of CARs with synthetic co-signaling modules adopted from immunoglobin-like superfamily (IgSF) and tumor necrosis factor receptor superfamily (TNFRSF). We quantitatively characterized the signaling behaviors of these recombinants by both NFAT and NF-κB reporter, and identified a set of new CARs with diverse signaling behaviors. Specifically, the 28(NM)-BB(MC) CAR-T cells exhibited improved cytotoxicity and T cell persistence. The synthetic approach can promote our understanding of the signaling principles of CAR molecule, and provide a powerful tool box for CAR-T cell engineering.

A deep learning model for lymph node metastasis prediction based on digital histopathological images of primary endometrial cancer.

Background: The current study aimed to develop a deep learning (DL) model for prediction of lymph node metastasis (LNM) based on hematoxylin and eosin (HE)-stained histopathological images of endometrial cancer (EC). The model was validated using external data. Methods: A total of 2,104 whole slide image (WSI) from 564 patients with pathologically confirmed LNM status were collated from West China Second University Hospital. An artificial intelligence (AI) model was built on the multiple instance-learning (MIL) framework for automatic prediction of the probability of LNM and its performance compared with "Mayo criteria". An additional external data source comprising 533 WSI was collected from two independent medical institutions to validate the model's robustness. Heatmaps were generated to demonstrate regions of the WSI that made the greatest contributions to the DL network output to improve understanding of these processes. Results: The proposed MIL model achieved an area under the curve (AUC) of 0.938, a sensitivity of 0.830 and a specificity of 0.911 for LNM prediction to EC. The AUC according to Mayo criteria was 0.666 for the same test dataset. For types I, II and mixed EC, AUCs were 0.927, 0.979 and 0.929, respectively. The predictive performance of the MIL model also achieved an AUC of 0.921 for early staging. In external validation data, the proposed model achieved an AUC of 0.770, a sensitivity of 0.814 and a specificity of 0.520 for LNM prediction. AUCs were 0.783 for type I and 0.818 for early stage EC. Conclusions: The proposed MIL model generated from histopathological images of EC has a much better LNM predictive performance than that of Mayo criteria. A novel DL-based biomarker trained on different histological subtypes of EC slides was revealed to predict metastatic status with improved accuracy, especially for early staging patients. The current study proves the concept of MIL-based prediction of LNM in EC for the first time, and brought a new sight to improve the accuracy of LNM prediction. Multicenter prospective validation data is required to further confirm the clinical utility.

Temporal transcriptomic changes in microRNAs involved in the host immune response and metabolism during Neospora caninum infection.

BACKGROUND: Neospora caninum infection is a major cause of abortion in cattle, which results in serious economic losses to the cattle industry. However, there are no effective drugs or vaccines for the control of N. caninum infections. There is increasing evidence that microRNAs (miRNAs) are involved in many physiological and pathological processes, and dysregulated expression of host miRNAs and the biological implications of this have been reported for infections by various protozoan parasites. However, to our knowledge, there is presently no published information on host miRNA expression during N. caninum infection. METHODS: The expression profiles of miRNAs were investigated by RNA sequencing (RNA-seq) in caprine endometrial epithelial cells (EECs) infected with N. caninum at 24 h post infection (pi) and 48 hpi, and the functions of differentially expressed (DE) miRNAs were predicted by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. The transcriptome data were validated by using quantitative real-time polymerase chain reaction. One of the upregulated DEmiRNAs, namely chi-miR-146a, was selected to study the effect of DEmiRNAs on the propagation of N. caninum tachyzoites in caprine EECs. RESULTS: RNA-seq showed 18 (17 up- and one downregulated) and 79 (54 up- and 25 downregulated) DEmiRNAs at 24 hpi and 48 hpi, respectively. Quantitative real-time polymerase chain reaction analysis of 13 randomly selected DEmiRNAs (10 up- and three downregulated miRNAs) confirmed the validity of the RNA-seq data. A total of 7835 messenger RNAs were predicted to be potential targets for 66 DEmiRNAs, and GO and KEGG enrichment analysis of these predicted targets revealed that DEmiRNAs altered by N. caninum infection may be involved in host immune responses (e.g. Fc gamma R-mediated phagocytosis, Toll-like receptor signaling pathway, tumor necrosis factor signaling pathway, transforming growth factor-ß signaling pathway, mitogen-activated protein kinase signaling pathway) and metabolic pathways (e.g. lysine degradation, insulin signaling pathway, AMP-activated protein kinase signaling pathway, Rap1 signaling pathway, calcium signaling pathway). Upregulated chi-miR-146a was found to promote N. caninum propagation in caprine EECs. CONCLUSIONS: This is, to our knowledge, the first report on the expression profiles of host miRNAs during infection with N. caninum, and shows that chi-miR-146a may promote N. caninum propagation in host cells. The novel findings of the present study should help to elucidate the interactions between host cells and N. caninum.

Dysregulation of Ceramide Metabolism Is Linked to Iron Deposition and Activation of Related Pathways in the Aorta of Atherosclerotic Miniature Pigs.

The miniature pig is a suitable animal model for investigating human cardiovascular diseases. Nevertheless, the alterations in lipid metabolism within atherosclerotic plaques of miniature pigs, along with the underlying mechanisms, remain to be comprehensively elucidated. In this study, we aim to examine the alterations in lipid composition and associated pathways in the abdominal aorta of atherosclerotic pigs induced by a high-fat, high-cholesterol, and high-fructose (HFCF) diet using lipidomics and RNA-Seq methods. The results showed that the content and composition of aortic lipid species, particularly ceramide, hexosyl ceramide, lysophosphatidylcholine, and triglyceride, were significantly altered in HFCF-fed pigs. Meanwhile, the genes governing sphingolipid metabolism, iron ion homeostasis, apoptosis, and the inflammatory response were significantly regulated by the HFCF diet. Furthermore, C16 ceramide could promote iron deposition in RAW264.7 cells, leading to increased intracellular reactive oxygen species (ROS) production, apoptosis, and activation of the toll-like receptor 4 (TLR4)/nuclear Factor-kappa B (NF-қB) inflammatory pathway, which could be mitigated by deferoxamine. Our study demonstrated that dysregulated ceramide metabolism could increase ROS production, apoptosis, and inflammatory pathway activation in macrophages by inducing iron overload, thus playing a vital role in the pathogenesis of atherosclerosis. This discovery could potentially provide a new target for pharmacological therapy of cardiovascular diseases such as atherosclerosis.

Elevating Photooxidation of Methane to Formaldehyde via TiO2 Crystal Phase Engineering.

Photocatalytic conversion of methane to value-added products under mild conditions, which represents a long sought-after goal for industrial sustainable production, remains extremely challenging to afford high production and selectivity using cheap catalysts. Herein, we present the crystal phase engineering of commercially available anatase TiO2 via simple thermal annealing to optimize the structure-property correlation. A biphase catalyst with anatase (90%) and rutile (10%) TiO2 with the optimal phase interface concentration exhibits exceptional performance in the oxidation of methane to formaldehyde under the reaction conditions of water solvent, oxygen atmosphere, and full-spectrum light irradiation. An unprecedented production of 24.27 mmol gcat-1 with an excellent selectivity of 97.4% toward formaldehyde is acquired at room temperature after a 3 h reaction. Both experimental results and theoretical calculations disclose that the crystal phase engineering of TiO2 lengthens the lifetime of photogenerated carriers and favors the formation of intermediate methanol species, thus maximizing the efficiency and selectivity in the aerobic oxidation of methane to formaldehyde. More importantly, the feasibility of the scale-up production of formaldehyde is demonstrated by inventing a "pause-flow" reactor. This work opens the avenue toward industrial methane transformation in a sustainable and economical way.

Case Report: Giant Cell Tumor of Tendon Sheath After Breast Augmentation.

Giant cell tumor of tendon sheath (GCTTS) is a benign tumor. It occurs predominantly in the hands, ankles, and knees. A 39-year-old female presented with GCTTS in the right breast after breast augmentation. There was a clear borderline between the tumor and breast tissue. In terms of morphological appearance, synovial metaplasia could be observed in part of the collagenous capsule. The tumor was moderately cellular and was composed of synovium-like monocytes. The main part of the tumor was blended with nested and scattered xanthomatous cells, lymphocytes, and osteoclast-like giant cells. Hemosiderin granules were distributed in the lesion. Immunohistochemical staining and fluorescence in situ hybridization (FISH) analyses were performed. CD68 staining was positive in osteoclast-like giant cells. In addition, neither significant USP6 translocation nor CSF1 translocation was detected by FISH. We hypothesized that the pathogenesis of this rare GCT-TS was based on synovial metaplasia and did not depend on the translocation of classical CSF1.

A Chimeric Conjugate of Antibody and Programmable DNA Nanoassembly Smartly Activates T Cells for Precise Cancer Cell Targeting.

Synthetically directing T-cells against tumors emerges as a promising strategy in immunotherapy, while it remains challenging to smartly engage T cells with tunable immune response. Herein, we report an intelligent molecular platform to engineer T-cell recognition for selective activation to potently kill cancer cells. To this end, we fabricated a hybrid conjugate that uses a click-type DNA-protein conjugation to equip the T cell-engaging antibody with two distinct programmable DNA nanoassemblies. By integrating multiple aptameric antigen-recognitions within a dynamic DNA circuit, we achieved combinatorial recognition of triple-antigens on cancer cells for selective T-cell activation after high-order logic operation. Moreover, by coupling a DNA nanostructure, we precisely defined the valence of the antigen-binding aptamers to tune avidity, realizing effective tumor elimination in vitro and in vivo. Together, we present a versatile and programmable strategy for synthetic immunotherapy.

Perspectives on Ligand Properties of N-Heterocyclic Carbenes in Iron Porphyrin Complexes.

There has been considerable research interest in the ligand nature of N-heterocyclic carbenes (NHCs). In this work, two six-coordinate NHC iron porphyrin complexes [FeII(TTP)(1,3-Me2Imd)2] (TTP = tetratolylporphyrin, 1,3-Me2Imd = 1,3-dimethylimidazol-2-ylidene) and [FeIII(TDCPP)(1,3-Me2Imd)2]ClO4 (TDCPP = 5,10,15,20-tetrakis(2,6-dichlorophenyl)porphyrin) are reported. Single-crystal X-ray characterizations demonstrate that both complexes have strongly ruffled conformations and relatively perpendicular ligand orientations which are forced by the sterically bulky 1,3-Me2Imd NHC ligands. Multitemperature (4.2-300 K) and high magnetic field (0-9 T) Mössbauer and low-temperature (4.0 K) EPR spectroscopies definitely confirmed the low-spin states of [FeII(TTP)(1,3-Me2Imd)2] (S = 0) and [FeIII(TDCPP)(1,3-Me2Imd)2]ClO4 (S = 1/2). The similarity of 1,3-Me2Imd and imidazole, as well as the well-established correlations between the ligand nature and spectroscopic characteristics of [FeII,III(Porph)(L)2]0,+ (Porph: porphyrin; L: planar base ligand) species, allowed direct comparisons between the pair of ligands which revealed for the first time that NHC has a stronger π-acceptor ability than imidazoles, in addition to its very strong σ-donation.

Clinicopathologic characteristics of HER2-positive metaplastic squamous cell carcinoma of the breast.

AIMS: The aim of this study was to analyse the clinicopathological features and prognosis of human epidermal growth factor receptor-2 (HER2)-positive metaplastic squamous cell carcinoma (MSCC). METHODS: Fifty-eight patients with MSCC of the breast who were classified into 45 triple-negative and 13 HER2-positive subgroups diagnosed at the West China Hospital, Sichuan University, from 2004 to 2018, were enrolled. Clinicopathological features were collected and compared between HER2-positive MSCC, triple-negative MSCC, HER2-positive invasive breast carcinoma of no special type (NST) and triple-negative NST groups. In the prognostic survival analysis, HER2-positive MSCCs was compared with triple-negative MSCCs, HER2-positive NSTs and triple-negative NSTs. RESULTS: Compared with triple-negative MSCCs, more patients with Ki-67 low expression were in HER2-positive MSCCs (p<0.05). More patients with HER2-positive MSCC than patients with HER2-positive NST were postmenopausal (p<0.05). Compared among HER2-positive MSCCs, triple-negative MSCCs and triple-negative NSTs, patients of HER2-positive MSCCs with high Ki-67 expression were the least, and HER2-positive MSCCs had more strongly associated with postmenopausal disease status (p<0.05). In survival analyses, HER2-positive MSCCs had a high risk of recurrence and poor prognosis (p<0.05). Lymph node status was significantly associated with the disease-free survival of patients with HER2-positive MSCC. CONCLUSION: In conclusion, our study indicates that HER2-positive MSCC is an aggressive disease with unique clinicopathological characteristics. Both HER2-positive status and an SCC component are critical factors for poor prognosis. HER2-positive MSCC and triple-negative MSCC are distinct subgroups. Corresponding targeted therapy recommendations should be made for this HER2-positive MSCC group.

Genistein reverses the effect of 17ß-estradiol on exacerbating experimental occlusal interference-induced chronic masseter hyperalgesia in ovariectomised rats.

BACKGROUND: Oro-facial pain is more prevalent in women than in men, and oestrogen may underlie this sex difference. Genistein reversed the potentiation of 17ß-estradiol (E2) on glutamate-induced acute masseter nociceptive behaviour, but its role in dental experimental occlusal interference (EOI)-induced chronic masseter hyperalgesia remains unclear. OBJECTIVE: This study aimed to investigate sex differences, and to explore the role and underlying mechanisms of genistein in E2-potentiated EOI-induced chronic masseter hyperalgesia in rats. METHODS: Female and male rats were prepared to compare the sex differences of masseter hyperalgesia induced by EOI using a 0.4-mm-thick metal crown. Female rats were ovariectomised (OVX) and treated with E2 and genistein, followed by EOI. The head withdrawal threshold (HWT) was examined to assess masseter sensitivity. The protein expression of transient receptor potential vanilloid-1 (TRPV1) in the trigeminal ganglion (TG) was detected using western blotting. Immunofluorescence staining was used to reveal the colocalisation of oestrogen receptors (ERs) with TRPV1 and the percentage of TRPV1-positive neurons in the TG. RESULTS: To some extent, female rats displayed enhanced sensitivity to EOI-induced chronic masseter hyperalgesia compared with males. Female rats showed the lowest HWT in the pro-oestrus phase. Pre-treatment with genistein antagonised E2 potentiation in EOI-induced masseter hyperalgesia and blocked the effect of E2 by downregulating TRPV1 protein expression and the percentage of TRPV1-positive neurons in the TG. CONCLUSION: Female rats showed greater masseter hyperalgesia than males under EOI. Genistein antagonised the facilitation of EOI-induced chronic masseter hyperalgesia by E2 probably through inhibiting TRPV1 in the TG.

AAV capsid variants with brain-wide transgene expression and decreased liver targeting after intravenous delivery in mouse and marmoset.

Genetic intervention is increasingly being explored as a therapeutic option for debilitating disorders of the central nervous system. The safety and efficacy of gene therapies rely upon expressing a transgene in affected cells while minimizing off-target expression. Here we show organ-specific targeting of adeno-associated virus (AAV) capsids after intravenous delivery, which we achieved by employing a Cre-transgenic-based screening platform and sequential engineering of AAV-PHP.eB between the surface-exposed AA452 and AA460 of VP3. From this selection, we identified capsid variants that were enriched in the brain and targeted away from the liver in C57BL/6J mice. This tropism extends to marmoset (Callithrix jacchus), enabling robust, non-invasive gene delivery to the marmoset brain after intravenous administration. Notably, the capsids identified result in distinct transgene expression profiles within the brain, with one exhibiting high specificity to neurons. The ability to cross the blood-brain barrier with neuronal specificity in rodents and non-human primates enables new avenues for basic research and therapeutic possibilities unattainable with naturally occurring serotypes.

Bioinformatics and Transcriptome Analysis of CFEM Proteins in Fusarium graminearum.

Fusarium blight of wheat is usually caused by Fusarium graminearum, and the pathogenic fungi will secrete effectors into the host plant tissue to affect its normal physiological process, so as to make it pathogenic. The CFEM (Common in Fungal Extracellular Membrane) protein domain is unique to fungi, but it is not found in all fungi. The CFEM protein contained in F. graminearum may be closely related to pathogenicity. In this study, 23 FgCFEM proteins were identified from the F. graminearum genome. Then, features of these proteins, such as signal peptide, subcellular localization, and transmembrane domains, etc., were analyzed and candidate effectors were screened out. Sequence alignment results revealed that each FgCFEM protein contains one CFEM domain. The amino acids of the CFEM domain are highly conserved and contain eight spaced cysteines, with the exception that FgCFEM8, 9, and 15 lack two cysteines and three cysteines were missed in FgCFEM18 and FgCFEM22. A recently identified CFEM_DR motif was detected in 11 FgCFEMs, and importantly we identified two new conserved motifs containing about 29 and 18 amino acids (CFEM_WR and CFEM_KF), respectively, in some of FgCFEM proteins. Transcriptome analysis of the genes encoding CFEM proteins indicated that all the CFEM-containing genes were expressed during wheat infection, with seven and six genes significantly up- and down-regulated, respectively, compared with in planta and in vitro. Based on the above analysis, FgCFEM11 and FgCFEM23 were predicted to be F. graminearum effectors. This study provides the basis for future functional analyses of CFEM proteins in F. graminearum.