ObjectiveTo investigate the expression of prefrontal cortical neurons， methyltransferase-like 3 （METTL3）， fat mass and obesity-associated gene （FTO）， and AlkB homolog 5 （ALKBH5） proteins in a mouse model of post-traumatic stress disorder （PTSD）. MethodsA PTSD mouse model was established using a single prolonged stress and foot shock stimulation （SPSS） method. The despair， anxiety， and learning and memory functions of PTSD mice were assessed through the open field test， Y-maze test， and forced swimming test. Neuronal damage was detected via HE and Nissl staining. The expression levels of METTL3， FTO， ALKBH5， and neuronal nuclear protein （NEUN） were assessed by Western blot and immunofluorescence staining. ResultsCompared to control group， PTSD mice subjected to SPSS exhibited signs of despair， anxiety， and impaired learning and memory. HE and Nissl staining results showed neuronal damage in the prefrontal cortex of PTSD mice. Western blot and immunofluorescence staining results showed that the expression of the m6A-related proteins METTL3 and FTO decreased， while the expression of ALKBH5 increased in the prefrontal cortex. Additionally， NEUN protein levels showed a declining trend. ConclusionThe pathogenesis of PTSD may be associated with neuronal damage in the prefrontal cortex and alterations in m6A methylation proteins.

ObjectiveTo investigate the effect of protein arginine methyltransferase 5 （PRMT5） on the angiogenesis of colorectal cancer （CRC） through the Janus kinase 2/signal transducer and activator of transcription 3 （JAK2/STAT3） signaling pathway. MethodsThe expression and prognostic value of PRMT5 in CRC tissues were analyzed based on the cancer genome atlas （TCGA） and gene expression omnibus （GEO） databases. A total of 105 pairs of cancer tissues and matched adjacent non-tumor tissues were collected from CRC patients， and PRMT5 expression was detected by immunohistochemistry （IHC）； Western blot was used to detect the PRMT5 protein level in normal colonic epithelial cells （FHC） and CRC cell lines （including RKO， SW480）. SW48 and LoVo cells were divided into negative control （NC） group and PRMT5 knockdown interference group. After the stable knockdown cell models were successfully constructed， the effects of conditioned medium on the proliferation， cell cycle， migration and tube formation abilities of human umbilical vein endothelial cells （HUVECs） were detected； the correlation between PRMT5 and vascular endothelial growth factor A （VEGFA） was analyzed by real-time quantitative polymerase chain reaction （RT-qPCR） and enzyme-linked immunosorbent assay （ELISA）. Gene set enrichment analysis （GSEA） and cytoplasm-nucleus separation assay were performed to verify the regulatory pathway. ResultsPRMT5 was highly expressed in CRC tissues and cell lines， which was significantly correlated with poor prognosis and pathological characteristics of patients （all P<0.05）； PRMT5 knockdown could significantly inhibit the angiogenesis-related abilities of HUVECs （all P<0.05） and down-regulate the expression of VEGFA （all P<0.01）. GSEA results indicated that PRMT5 expression was significantly associated with the activation of JAK2/STAT3 signaling pathway （all P<0.001）. PRMT5 knockdown could inhibit the activation of JAK2/STAT3， the nuclear translocation of STAT3 and the protein level of phosphorylated STAT3 （p-STAT3） in the nucleus， thus inhibiting the expression of VEGFA. ConclusionPRMT5 is highly expressed in CRC tissues and indicates poor prognosis of patients. It promotes the nuclear translocation and transcriptional activity of STAT3 by activating the JAK2/STAT3 pathway and drives the expression of VEGFA to promote tumor angiogenesis.

In recent years, the spread of clubroot disease caused by Plasmodiophora brassicae infection has seriously affected the yield and quality of Brassica juncea (L.) Czern.. The cascade of mitogen-activated protein kinases (MAPKs), a highly conserved signaling pathway, plays an important role in plant responses to both biotic and abiotic stress conditions. To mine the MAPK genes related to clubroot disease resistance in B. juncea, we conducted a genome-wide analysis on this vegetable, and we analyzed the phylogenetic evolution and gene structure of the MAPK gene family in mustard. The 66 BjuMAPK genes identified by screening the whole genome sequence of B. juncea were unevenly distributed on 17 chromosomes. At the genomic scale, tandem repeats led to an increase in the number of MAPK genes in B. juncea. It was found that members of the same subfamily had similar gene structures, and there were great differences among different subfamilies. These predicted cis-acting elements were related to plant hormones, stress resistance, and plant growth and development. The expression of BjuMAPK02, BjuMAPK15, BjuMAPK17, and BjuMAPK19 were down-regulated or up-regulated in response to P. brassicae infection. The above results lay a theoretical foundation for further studying the functions of BjuMAPK genes in B. juncea in response to the biotic stress caused by clubroot disease.
Mustard Plant/parasitology* ; Plasmodiophorida/pathogenicity* ; Plant Diseases/genetics* ; Mitogen-Activated Protein Kinases/metabolism* ; Phylogeny ; Disease Resistance/genetics* ; Gene Expression Regulation, Plant ; Genome, Plant ; Plant Proteins/genetics*

Phage immunoprecipitation sequencing (PhIP-Seq) is a high-throughput and low-cost method for analyzing the specific binding of target proteins to peptide libraries. The method uses oligonucleotide library synthesis (OLS) to encode proteome-scale peptide libraries for display on phages, and then immunoprecipitates these library phages with target proteins (such as antibodies) for subsequent analysis by high-throughput DNA sequencing. PhIP-Seq enables the screening of peptide targets that react specifically with hundreds of proteins or pathogens. PhIP-Seq has been successfully applied in various fields such as disease detection, screening of autoimmune disease biomarkers, vaccine development, and allergen detection, becoming a high-throughput diagnostic technology. This article systematically describes the development, applications, and result evaluation of PhIP-Seq, in order to gain a more comprehensive understanding of the application and future development prospects of this technology in various fields.
Peptide Library ; Humans ; Immunoprecipitation/methods* ; High-Throughput Nucleotide Sequencing/methods* ; Bacteriophages/genetics*

Objective:To explore the expression changes of N6-methyladenosine (m6A)-related proteins in the hippocampus of mice with post traumatic stress disorder (PTSD)-like behavior and the therapeutic effects of sertraline hydrochloride.Methods:Male C57BL/6J mice aged 4-6 weeks were selected to establish a PTSD model using a single prolonged stress and foot shock stimulation. A total of 24 mice were randomly divided into the control group, model group, and sertraline group using a random number table, with 8 mice in each group. Mice in the sertraline group were intraperitoneally injected with sertraline hydrochloride (15 mg/kg, once daily) 24 h after PTSD modelling, continuing for 14 days. Mice in the control group and model group were injected with an equal volume of 0.9% NaCl solution (once daily, for 14 days). The anxiety, despair, and learning and memory functions of the mice were assessed using the open field test, Y-maze test, and forced swimming test. Western blot was performed to measure the protein expression levels of methyltransferase-like protein 3 (METTL3), fat mass and obesity-associated gene (FTO), ALKB homolog 5 (ALKBH5), Wilms tumour 1 associating protein (WTAP), and methyltransferase-like protein 14 (METTL14) in the hippocampus. Immunofluorescence was used to detect the expression levels of METTL3, FTO, and ALKBH5 in the hippocampus. Statistical analysis was performed using SPSS 26.0 and GraphPad Prism 9.0.Comparisons between two groups were conducted using independent samples t-test, while comparisons among three groups were performed using one-way analysis of variance (ANOVA) or Kruskal-Wallis H test, followed by pairwise comparisons using LSD test. Results:(1) Behavioral results showed that the total distance travelled in the central area ( F=9.231, P<0.05) and the time spent in the central area ( H=8.045, P<0.05) showed statistically significant differences among the control, model, and sertraline groups. Mice in the control and sertraline groups travelled a greater distance((332.68±121.17)cm, (248.56±40.21)cm) and spent more time(24.98(23.08, 26.71)s, 22.52(18.86, 26.20)s) in the central area than those in the model group((131.66±84.90)cm, 9.14(6.56, 18.53)s) (all P<0.05). In the forced swimming test, the number of resting episodes ( F=16.882, P<0.05) and the duration of rest ( H=12.285, P<0.05) differed significantly among the three groups. Mice in the control group ((19.14±8.30) counts, 30.21 (18.98, 52.62) s) and the sertraline group ((17.63±8.14) counts, 25.90 (16.78, 37.56) s) had fewer resting episodes and shorter resting durations compared to those in the model group ((37.75±6.47) counts, 83.37 (64.62, 124.42) s) (all P<0.05). The percentage of alternations in the Y-maze experiment showed significant statistical differences among the three groups( F=6.844, P<0.05). Mice in the control group ((51.33±11.49)%) and the sertraline group ((48.24±3.10)%) exhibited a higher percentage of alternations than that in the model group ((36.70±8.15)%) ( P<0.05). (2) Western blot results showed that the protein expression levels of METTL3, FTO, and ALKBH5 in the hippocampal tissue of the three groups showed significant differences ( F=10.263, 9.010, 6.950, all P<0.05). The METTL3 and FTO protein expression levels in the hippocampus in the control group (0.85±0.07, 0.86±0.04) and the sertraline group (0.93±0.06, 0.95±0.13) were higher than those in the model group (0.74±0.02, 0.68±0.04) (all P<0.05). However, the ALKBH5 protein expression levels in the control group (0.93±0.08) and the sertraline group (0.87±0.13) were lower than that in the model group (1.13±0.04) (both P<0.05). (3) Immunofluorescence results showed that the expression levels of METTL3, FTO, and ALKBH5 proteins in the hippocampal tissue of the three groups showed significant statistical differences ( F=37.912, 62.659, 54.417, all P<0.05). The expression levels of METTL3 and FTO in the hippocampus in the control group (14.03±0.32, 13.85±0.28) and the sertraline group (17.94±0.29, 10.52±0.66) were higher than those in the model group (11.67±1.48, 8.70±0.68) (all P<0.05). The expression levels of ALKBH5 in the control group (12.94±0.38) and the sertraline group (13.30±0.93) were lower than that in the model group (19.24±1.03) (both P<0.05). Conclusion:The expression of m6A-related proteins in the hippocampus of PTSD-like mice is altered. Sertraline treatment can significantly regulate the expression of these proteins and improve anxiety, despair, and learning and memory impairments in the PTSD-like mice.

Objective:To explore changes in brain activity in patients with post-traumatic stress disorder(PTSD).Methods:A total of 40 participants involved in car accidents were included,and functional magnetic reso-nance imaging(fMRI)scans were collected within one week.Anxiety,depression,and personality assessments were conducted with the Hamilton Anxiety Scale(HAMA),Hamilton Depression Scale(HAMD),and Eysenck Person-ality Scale for Adult(EPQ).After two months,a second fMRI scan was conducted,and a PTSD diagnosis was made.Participants were divided into a trauma-exposed group(n=23)and a PTSD group(n=17)based on wheth-er they developed PTSD.Changes in brain functional activity between the trauma-exposed group and the PTSD group were compared using the percentage of amplitude fluctuation(perAF)method.Results:Compared to the trauma-exposed group,the PTSD group showed a decreased perAF value in the left hippocampus at 1 week,and de-creased perAF values in the right mid-cingulate gyrus and left postcentral gyrus at 2 months(P＜0.05).When comparing the PTSD group at different times,the perAF values in the left middle temporal gyrus and left medial su-perior frontal gyrus decreased at 2 months(P＜0.05).Correlation analysis revealed that PCL-5 scores were posi-tively correlated with EPQ Psychoticism(r=0.32,P=0.041),HAMA(r=0.35,P＜0.05),and HAMD(r=0.34,P＜0.05).Regression analysis found that higher scores of EPQ psychoticism(OR=11.79)and HAMA(OR=1.62)were risk factors for post-accident PTSD,while higher scores of EPQ extraversion(OR=0.32)were pro-tective factors.Conclusion:It suggests that patients with post-traumatic stress disorder may show decreased activity in the right middle cingulate cortex,left postcentral gyrus,left middle temporal gyrus,and left medial superior fron-tal gyrus within two months after the traumatic event.

Allergen-specific immunotherapy（AIT） remains the only therapeutic approach with the potential to modify the natural course of allergic rhinitis（AR）. Nevertheless, considerable inter-individual variability exists in patients'responses to AIT. To facilitate more reliable assessment of treatment efficacy, the China Rhinopathy Research Cooperation Group（CRRCG） convened young and middle-aged nasal experts in China to formulate the present consensus. The recommended subjective outcome measures for AIT comprise symptom scores, medication scores, combined symptom and medication scores, quality-of-life assessments, evaluation of disease control, and assessment of comorbidities. Objective indicators may supplement these measures. Currently available objective approaches include skin prick testing, nasal provocation testing, and allergen exposure chambers. However, these methods remain constrained by practical limitations and are not yet appropriate for routine implementation in clinical efficacy evaluation. In addition, several biomarkers, including sIgE and the sIgE/tIgE ratio, sIgG4, serum IgE-blocking activity, IgA, cytokines and chemokines, as well as immune cell surface molecules and their functional activity, have been shown to have associations with AIT outcomes. While these biomarkers may complement subjective assessments, they are subject to significant limitations. Consequently, large-scale multicenter trials and real-world evidence are required to strengthen the evidence base. The present consensus underscores the necessity of integrating patients'subjective experiences with objective testing throughout the treatment process, thereby providing a more comprehensive and accurate framework for efficacy evaluation. Looking forward, future investigations should prioritize the incorporation of multi-omics data and artificial intelligence methodologies, which hold promise for overcoming current limitations in assessment strategies and for advancing both the standardization and personalization of AIT.
Humans ; Allergens/immunology* ; China ; Consensus ; Desensitization, Immunologic ; Immunoglobulin E ; Quality of Life ; Rhinitis, Allergic/therapy* ; Treatment Outcome ; East Asian People

Sonodynamic therapy (SDT) can potentially induce immunogenic cell death in tumor cells, leading to the release of ATP, and facilitating the initiation of an immune response. Nevertheless, the enzymes CD39 and CD73 can swiftly convert ATP into immunosuppressive adenosine (ADO), resulting in an immunosuppressive tumor microenvironment (TME). This study introduced a nanomedicine (QD/POM1@NP@M) engineered to reprogram TME by modulating the CD39/CD73/ADO pathway. The nanomedicine encapsulated sonosensitizers silver sulfide quantum dots, and the CD39 inhibitor POM1, while also incorporating homologous tumor cell membranes to enhance targeting capabilities. This integrated approach, on the one hand, stimulates the release of ATP via SDT, thereby initiating the immune response. In addition, it reduced the accumulation of ADO by inhibiting CD39 activity, which ameliorated the immunosuppressive TME. Upon administration, the nanomedicine demonstrated substantial anti-tumor efficacy by facilitating the infiltration of anti-tumor immune cells, while reducing the immunosuppressive cells. This modulation effectively transformed the TME from an immunologically "cold" state to a "hot" state. Furthermore, combined with the checkpoint inhibitor α-PDL1, the nanomedicine augmented systemic anti-tumor immunity and promoted the establishment of long-term immune memory. This study provides an innovative strategy for combining non-invasive SDT and ATP-driven immunotherapy, offering new ideas for future cancer treatment.

Temporal interference (TI) is a form of stimulation that epitomizes an innovative and non-invasive approach for profound neuromodulation of the brain, a technique that has been validated in mice. Yet, the thin cranial bone structure of mice has a marginal influence on the effect of the TI technique and may not effectively showcase its effectiveness in larger animals. Based on this, we carried out TI stimulation experiments on rats. Following the TI intervention, analysis of electrophysiological data and immunofluorescence staining indicated the generation of a stimulation focus within the nucleus accumbens (depth, 8.5 mm) in rats. Our findings affirm the viability of the TI methodology in the presence of thick cranial bones, furnishing efficacious parameters for profound stimulation with TI administered under such conditions. This experiment not only sheds light on the intervention effects of TI deep in the brain but also furnishes robust evidence in support of its prospective clinical utility.
Animals ; Deep Brain Stimulation/methods* ; Nucleus Accumbens/physiology* ; Male ; Rats ; Rats, Sprague-Dawley ; Time Factors