Periodontitis is an infectious and inflammatory disease. The inability to control clinical inflammation significantly contributes to unsuccessful periodontal treatment. As a key regulator of epigenetics, Sirtuin 6 is one of the NAD+-dependent histone deacetylase family members and has recently gained considerable interest in the context of inflammation. While MDL-800 has been demonstrated as an activator of SIRT6, its function in treating periodontitis remains unknown. In this study, animal experiments revealed that MDL-800 reduced alveolar bone resorption in ligature-induced periodontitis, particularly via reducing inflammation and inhibiting osteoclast production in a SIRT6-dependent manner. Furthermore, human gingival fibroblasts (HGFs) were exposed to lipopolysaccharide (LPS) to mimic periodontal inflammation in vitro. We noted that MDL-800 remarkably reduced expressions of pro-inflammatory cytokines in LPS-induced HGFs, which was neutralized by inhibiting SIRT6. Besides, MDL-800 could also reduce oxidative stress damage under inflammatory conditions. The suppression of AIM2 inflammasome activation was associated with the significant anti-inflammatory properties of MDL-800, as demonstrated by RNA sequencing and validation experiments. In conclusion, these findings suggest that MDL-800 was effective in reducing periodontal inflammation, partly by inhibiting AIM2 pathway in a SIRT6-dependent manner, demonstrating significant promise for enhancing therapeutic outcomes in periodontitis.

Sepsis-associated encephalopathy (SAE) is a severe and common neurological complication of sepsis, characterized by symptoms ranging from mild confusion, delirium, deep coma, and severe cognitive dysfunction. Previous epidemiological and bioinformatics studies have revealed that HLA DP and DRA molecule play a pivotal role during sepsis. However, the mechanism by which these class II molecule contribute to cognitive impairment in SAE remains unclear. using the peritoneal contamination and infection model (PCI) model in humanized transgenic HLA-DP401/DRA-IAβ-/- genotypes mice, we aimed to investigate the effects of HLA class II haplotypes/alleles on sepsis and elucidate the underlying mechanism leading to cognitive impairment. Our results indicated that the introduction of HLA DP/DRA molecule significantly increased mortality, exacerbated clinical symptoms, and elevated inflammatory cytokine responses in both serum and hippocampal tissue of septic mice. Cecal slurry (CS) injection induced robust microglia activation and severe pathological damage of hippocampus. Furthermore, transcriptome analysis revealed numerous differentially expressed genes (DEGs) and prominent mitochondrial dysfunction in HLA-DP/DRA-IAβ-/- mice subjected to PCI. Notably, CS injection up-regulated AMPK-α phosphorylation in IAβ-/- mice but not in HLA DP/DRA-IAβ-/- mice. Consistently, sepsis induced persistent neurocognitive deficits and long-term anxiety-like behaviors in HLA DP/DRA-IAβ-/- PCI mice. In conclusion, these data provide direct evidence that HLA class II molecules modulate the host response to sepsis and highlight a critical role of HLA-DP/DRA in exacerbating the severity of systemic infection. The introduction of the HLA-DP and HLA-DRA genes synergistically upregulated systemic and hippocampal inflammatory cytokines, worsened clinical outcomes, impaired memory performance, and exacerbated long-term anxiety-like behaviors.

Bitter taste receptors (TAS2Rs), initially identified for chemosensory roles in the tongue, are expressed in extraoral tissues, including the airways. However, to date, it remains unclear whether bitter signaling is associated with susceptibility to bacterial infection in the lower airways and whether bitter signaling actually participates in the immune response in lung infection has yet to be genetically established. Here, we investigated the role of TAS2R signaling in Staphylococcus aureus-induced murine pneumonia via wild-type (WT) and several mutants (mTas2r104-/-/105-/-, mTas2r105-/-/114-/-, mTas2r104-/-/105-/-/114-/-, Gnat3-/- and Gnat3-/-mTas2r104-/-/105-/-) mice. Genetic disruption of TAS2Rs altered compensatory expression of other bitter receptors in the trachea and lungs, but did not affect immune cell composition in the lungs or thymus. Bitter receptor-deficient mice exhibited exacerbated pulmonary lesions at day 3 (D3) post-infection. Pulmonary infection significantly upregulated mTas2r105,106, 107, 108, 126, 136, 138 and Gnat3 in the lung. TAS2R signaling deficiency downregulated the expression of cytokines (e.g., IL-10, MIP-2) and antimicrobial peptides in the lungs and trachea, increased CD68+ macrophages in D3 lung tissues, amplified Ki67+ cell proliferation in alveolar and bronchiolar regions, and even impaired recovery from lung injury by day 14 (D14). Mechanistically, bitter taste pathway disruption dysregulated the mTOR pathway, reduced eNOS expression, and delayed resolution of pneumonia-induced injury. In summary, the current results collectively indicate that bitter taste signaling can modulate innate immune and inflammatory responses during S. aureus-induced lung infection.

Recently, the expression of bitter taste receptors and their downstream taste signaling cascade has been widely found outside the gustatory system, indicating important physiological functions of bitter taste receptors in various extraoral organs, including testis, but little is known about their functions in spermatogenesis. Here, we describe the localization and expression pattern of taste signaling transduction molecules in the testis. Genetic mutation of bitter signaling transduction molecules decreased the litter size, the IVF rate, and the diameter of seminiferous tubules and even resulted in empty seminiferous tubules. Transmission electron microscopy observations further revealed that overdeveloped acrosomes adhered to atrophic round spermatids in double-mutant mice. Mutant mice lacking bitter taste receptor signaling exhibited a dysfunction of adenosine 5'-monophosphate-activated protein kinase and inducible nitric oxide synthase signaling pathways, and a decreased expression of zonula occludens-1 and β-catenin in testis, indicating a disruption for the structure and functions of the blood-testis barrier. Transcriptome analysis further showed that bitter taste signaling deficiency can alter the expression profile of transcripts related to signal pathways, hormone synthesis, cell adhesion molecules, the chemokine signaling pathway, and cell metabolism in the testis, which finally contribute to impaired male fertility. In short, our work provides previously unidentified in vivo evidence that bitter taste signaling plays a critical role in the maintenance of normal spermatogenesis. These data further support the concept that bitter taste receptors exert functions outside the gustatory system and may have implications for the diagnosis and management of human male infertility.

Inflammatory bowel disease (IBD) is an immune-mediated gastrointestinal disorder that significantly impacts the life quality of people worldwide. Genetic factors play crucial roles in the development of IBD. Tas2rs, members of the G protein-coupled receptor (GPCR) superfamily, are known for their roles in bitter taste perception. However, Tas2rs have also been identified in the gut, where they help sense luminal contents and regulate gastrointestinal hormones. Periodontal Tas2r105 has been shown to modulate innate immunity by interacting with metabolites produced by oral bacteria. In this study, we observed increased Tas2r105 in the inflammatory colons induced by dextran sulfate sodium salt (DSS). We also noted that α-gustducin, the α-subunit of GPCRs, is present in the intestine, and that α-gustducin knockout mice exhibit aggravated colitis. Based on these findings, we hypothesize that Tas2r105 may play a role in immune regulation during IBD pathogenesis. To test this hypothesis, we used Tas2r105 knockout (KO) mice in a colitis model. Our results show that the KO mice had significantly shorter colon length, more severe colon inflammation, and greater destruction of the gut barrier compared with control mice. We also observed increased recruitment of macrophages to the lamina propria mucosa in the KO mice. Microbiological analysis revealed a significant increase in Proteobacteria and Bacteroidota, with a concomitant decrease in Firmicutes after Tas2r105 knockout. Metabolomic analysis showed a significant reduction in lysophosphatidylethanolamine (LPE) levels in the KO mice, which is known to have anti-inflammatory effects. Based on these findings, we speculate that Tas2r105 may help protect the intestine from inflammation by influencing the gut microbiota composition and LPE production.IMPORTANCEIncreased Tas2r105 was detected in the inflamed colon of mice outside the tongue. Tas2r105 deletion aggravated mice colon colitis. Tas2r105 might alleviate mice colitis by downregulating the Proteobacteria and the Bacteroidota abundance in the colon. Lysophosphatidylethanolamine (LPE) might be the key metabolite that mediated the intestinal protection of Tas2r105.

Background: Autism spectrum disorder (ASD) involves complex interactions between genetic and environmental factors. Recent studies suggest that dysregulation of β-arrestin2 (Arrb2) in the central nervous system is linked to ASD. However, its specific mechanisms remain unknown. Methods: This study employs a systems genetics approach to comprehensively investigate Arrb2 in multiple brain tissues, including the amygdala, cerebellum, hippocampus, and prefrontal cortex, using BXD recombinant inbred (RI) strains. In addition, genetic variance analysis, correlation analysis, expression quantitative trait loci (eQTL) mapping, and functional annotation were used to identify the key downstream targets of Arrb2, validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blotting (WB). Results: Arrb2 exhibited expression variations across the four brain regions in BXD mice. eQTL mapping revealed that Arrb2 is cis-regulated, and increased Arrb2 expression levels were significantly correlated with ASD-like symptoms, such as impaired social interactions and abnormal learning and memory. Furthermore, protein–protein interaction (PPI) network analysis, tissue correlation, functional relevance to autism, and differential expression identified eight downstream candidate genes regulated by Arrb2. The experimental results demonstrated that deletion of Arrb2 led to the downregulation of Myh9, Dnmt1, and Brd4 expression, along with protein kinase A (PKA)-induced hyperactivation of Synapsin I. These findings suggest that Arrb2 may contribute to the pathogenesis of autism by modulating the expression of these genes. Conclusions: This study highlights the role of Arrb2 in ASD pathogenesis and identifies Myh9, Dnmt1, and Brd4 as key downstream regulators. These findings provide new insights into the molecular mechanisms of ASD and pave the way for novel therapeutic targets.

Hepatocellular carcinoma (HCC) is the most common form of primary liver cancer with poor prognosis. Sorafenib, a first-line treatment for advanced HCC, has shown limited clinical benefits due to the onset of drug resistance. Thus, it is imperative to comprehend the mechanisms underlying sorafenib resistance and explore strategies to overcome or delay it. Here, we established HCC patient-derived xenograft (PDX) models with acquired resistance to sorafenib, and performed comprehensive proteomic and phosphoproteomic analyses on these models. The active cell cycle pathway along with the active cyclin-dependent kinase CDK1 and DNA-dependent protein kinase PRKDC was identified through KEGG pathway enrichment and kinase substrate enrichment analyses. Upon investigating the potential of combining sorafenib with putative kinase inhibitors, we found that the combination displays synergistic anti-proliferative effects in the sorafenib-resistant liver cancer cell line, thus providing a proof-of-concept for phosphoproteomic-guided design of precision medicine.

As a leaf vegetable, Gynura bicolor DC ( G. bicolor ) experiences a rapid deterioration after harvest including insufficient supply of sugar and destruction of cell membranes. In this research, four treatments were experimented on G. bicolor including the control (CK), 12% (g/g) sucrose (ST), 10 mu L L -1 1-MCP (MT), and the combination of sucrose and 1-MCP (SMT). The results showed that three treated groups reduced respiratory rate, inhibited hexose consumption and promoted the decrease of starch and sucrose, which was converted into hexose including glucose and fructose to maintain cell membrane integrity. Meanwhile, the activities of AI, NI, SS-C, amylase, and corresponding gene expression levels were significantly up-regulated in three treated groups at 1 d, among which AI played a crucial role in regulating the accumulation of hexose. Furthermore, ST exerted a pronounced effect on hexose accumulation at the beginning while MT reduced hexose consumption through lowered respiratory metabolism during storage. Notably, SMT exhibited an optimum preservation effect on inhibited respiratory metabolism, maintaining cell membrane integrity, enhancing the retention of hexose, indicating that a synergistic effect of ST and MT were developed during storage.

BACKGROUND:Staphylococcus aureus can cause serious infections by secreting many superantigen exotoxins in "carrier" or "pathogenic" states. HLA DQ and HLA DR humanized mice have been used as a small animal model to study the role of two molecules during S. aureus infection. However, the contribution of HLA DP to S. aureus infection is unknown yet. METHODS:In this study, we have produced HLA DP401 and HLA DRA0101 humanized mice by microinjection of C57BL/6J zygotes. Neo-floxed IAβ+/- mice were crossbred with Ella-Cre and further crossbred with HLA DP401 or HLA-DRA0101 humanized mice. After several rounds of traditional crossbreeding, we finally obtained HLA DP401-IAβ-/- and HLA DRA-IAβ-/- humanized mice, in which human DP401 or DRA0101 molecule was introduced into IAβ-/- mice deficient in endogenous murine MHC class II molecules. A transnasal infection murine model of S. aureus pneumonia was induced in the humanized mice by administering 2 × 108  CFU of S. aureus Newman dropwise into the nasal cavity. The immune responses and histopathology changes were further assessed in lungs in these infected mice. RESULTS:We evaluated the local and systemic effects of S. aureus delivered intranasally in HLA DP401-IAβ-/- and HLA DRA-IAβ-/- transgenic mice. S. aureus Newman infection significantly increased the mRNA level of IL 12p40 in lungs in humanized mice. An increase in IFN-γ and IL-6 protein was observed in HLA DRA-IAβ-/- mice. We observed a declining trend in the percentage of F4/80+ macrophages in lungs in HLA DP401-IAβ-/- mice and a decreasing ratio of CD4+ to CD8+ T cells in lungs in IAβ-/- mice and HLA DP401-IAβ-/- mice. A decreasing ratio of Vβ3+ to Vβ8+ T cells was also found in the lymph node of IAβ-/- mice and HLA DP401-IAβ-/- mice. S. aureus Newman infection resulted in a weaker pathological injury in lungs in IAβ-/- genetic background mice. CONCLUSION:These humanized mice will be an invaluable mouse model to resolve the pathological mechanism of S. aureus pneumonia and study what role DP molecule plays in S. aureus infection.

COVID-19 is caused by coronavirus SARS-CoV-2. Current systemic vaccines generally provide limited protection against viral replication and shedding within the airway. Recombinant VSV (rVSV) is an effective vector which inducing potent and comprehensive immunities. Currently, there are two clinical trials investigating COVID-19 vaccines based on VSV vectors. These vaccines were developed with spike protein of WA1 which administrated intramuscularly. Although intranasal route is ideal for activating mucosal immunity with VSV vector, safety is of concern. Thus, a highly attenuated rVSV with three amino acids mutations in matrix protein (VSVMT) was developed to construct safe mucosal vaccines against multiple SARS-CoV-2 variants of concern. It demonstrated that spike protein mutant lacking 21 amino acids in its cytoplasmic domain could rescue rVSV efficiently. VSVMT indicated improved safeness compared with wild-type VSV as the vector encoding SARS-CoV-2 spike protein. With a single-dosed intranasal inoculation of rVSVΔGMT-SΔ21, potent SARS-CoV-2 specific neutralization antibodies could be stimulated in animals, particularly in term of mucosal and cellular immunity. Strikingly, the chimeric VSV encoding SΔ21 of Delta-variant can induce more potent immune responses compared with those encoding SΔ21 of Omicron- or WA1-strain. VSVMT is a promising platform to develop a mucosal vaccine for countering COVID-19.

Background Bitter taste receptors (Tas2rs) are generally considered to sense various bitter compounds to escape the intake of toxic substances. Bitter taste receptors have been found to widely express in extraoral tissues and have important physiological functions outside the gustatory system in vivo.Methods To investigate the physiological functions of the bitter taste receptor cluster Tas2r106/Tas2r104/Tas2r105/Tas2r114 in lingual and extraoral tissues, multiple Tas2rs mutant mice and Gnat3 were produced using CRISPR/Cas9 gene-editing technique. A mixture containing Cas9 and sgRNA mRNAs for Tas2rs and Gnat3 gene was microinjected into the cytoplasm of the zygotes. Then, T7EN1 assays and sequencing were used to screen genetic mutation at the target sites in founder mice. Quantitative real-time polymerase chain reaction (qRT-PCR) and immunostaining were used to study the expression level of taste signaling cascade and bitter taste receptor in taste buds. Perception to taste substance was also studied using two-bottle preference tests.Results We successfully produced several Tas2rs and Gnat3 mutant mice using the CRISPR/Cas9 technique. Immunostaining results showed that the expression of GNAT3 and PLCB2 was not altered in Tas2rs mutant mice. But qRT-PCR results revealed the changed expression profile of mTas2rs gene in taste buds of these mutant mice. With two-bottle preference tests, these mutant mice eliminate responses to cycloheximide due to genetic mutation of Tas2r105. In addition, these mutant mice showed a loss of taste perception to quinine dihydrochloride, denatonium benzoate, and cucurbitacin B (CuB). Gnat3-mediated taste receptor and its signal pathway contribute to CuB perception.Conclusions These findings implied that these mutant mice would be a valuable means to understand the biological functions of TAS2Rs in extraoral tissues and investigate bitter compound-induced responses mediated by these TAS2Rs in many extraoral tissues.

The cytoplasmic multifunctional adaptor protein β-arrestin 2 (Arrb2) is involved in the occurrence of various nervous system diseases, such as Alzheimer's disease and Parkinson's disease. Previous laboratory studies have shown that the expression and function of the Arrb2 gene was increased in valproic acid-induced autistic mice models. However, few reports have examined the possible role of Arrb2 in the pathogenesis of autism spectrum disorder. Therefore, Arrb2-deficient (Arrb2-/-) mice were further studied to uncover the physiological function of Arrb2 in the nervous system. In this study, we found that Arrb2-/- mice had normal behavioral characteristics compared with wild-type mice. The autophagy marker protein LC3B was decreased in the hippocampus of Arrb2-/- mice compared to wild-type mice. Western blot analysis revealed that deletion of Arrb2 caused hyperactivation of Akt-mTOR signaling in the hippocampus. In addition, abnormal mitochondrial dysfunction was observed in Arrb2-/- hippocampal neurons, which was characterized by a reduction in mitochondrial membrane potential and adenosine triphosphate production and an increase in reactive oxygen species levels. Therefore, this study elucidates the interaction between Arrb2 and the Akt-mTOR signaling pathway and provides insights into the role of Arrb2 in hippocampal neuron autophagy.

Background Patellofemoral pain (PFP) is a common overuse injury among runners. It is not only a hindrance to the runner’s training, but also to the runner’s quality of life. PFP runners may strategize different running strategies to reduce patellofemoral joint stress, release pain, and improve function. Purpose This study aimed to determine the changes in joint coordination and variability under combinations of foot strike pattern and cadence for runners with patellofemoral pain. Methods Twenty male runners with PFP performed six running strategies which were two strike patterns named forefoot (FFS) and rearfoot (RFS) accompanied by three running cadences named slow10%, normal, and fast10%. A modified vector coding technique and circular statistics were respectively used to identify the coordination pattern and variability between hip sagittal-knee frontal (HsKf), hip sagittal-knee sagittal (HsKs) and knee transverse-ankle frontal (KtAf) during stance phase. Coordination patterns which were conformed with anatomical motion pattern was classified as mechanically sound, and the distribution frequency of each coordination pattern was quantified. Results Switching to FFS, the HsKf couples (p < 0.001, ES = 1.34) and the HsKs couples (p = 0.001, ES = 0.82) displayed significantly greater frequency in mechanically unsound coordination pattern during the initial stance phase. The effect of increasing running cadence on RFS displayed significantly greater frequency in mechanically unsound hip dominancy (p = 0.042, ES = 0.65) and knee dominancy (p = 0.05, ES = 0.70) coordination patterns for HsKf couples as well as for HsKs couples (p = 0.023, ES = 0.86) during the initial stance phase. Combined with FFS and fast10% cadence, HsKs couples showed more hip-dominated mechanical sound coordination pattern (p = 0.002, ES = 1.25). Further, altering footstrike pattern and cadence failed to change the coordination variability. Conclusions Changing running cadence (± 10%) combined with transfer strike pattern from RFS to FFS could not increase the distribution frequency in mechanically sound coordination patterns and change coordination variability for PFP runners.

Autism is a complex neurodevelopmental disease that may be caused by genetic and environmental factors, that are incompletely understood. Overactivation of dopaminergic receptors can lead to autistic-like behavior. β-arrestin2 (Arrb2) is a scaffolding protein of the arrestin family, which function as cytosolic multifunctional adapter proteins that activate cell signal transduction and mediate the signal termination and endocytosis of G-protein-coupled receptors (GPCRs) complexes. In this study, we established an Arrb2 knockout (Arrb2-/-) mouse to explore the biological function of Arrb2 in autistic-like behavior caused by abnormality in the dopaminergic system. We found that Arrb2-/- mice did not exhibit the autistic-like behavior normally induced by SKF38393, an agonist of the dopamine receptor 1 (D1R). Compared with wild-type (WT) untreated mice, the SKF38393-treated WT mice and Arrb2-/- mice, with or without SKF38393 treatment, showed abnormalities on electroencephalography (EEG) and increased stimulation of the phosphorylated form of extracellular signal-regulated kinase (p-ERK) via the PKA/Rap1/B-Raf/MEK pathway. These results demonstrated that Arrb2 regulated the dopaminergic system through the ERK signaling pathway in the occurrence and development of autism, and that targeted deletion of Arrb2 impeded the development of autistic-like behavior.

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This mixed methods study examines whether veteran–novice mentorship between tutors, as part of continuous in-service professional development, would have a positive effect on either party's transferable skills (e.g., communication, collaboration, and professionalism). Quantitative findings from pre- and postsurveys about the veteran–novice mentorship suggest that tutors have significant gains in some transferable skills, such as oral/written communication skills, teamwork/collaboration skills, digital technology skills, and career management skills, after attending the continuous in-service professional development. Quantitative findings from the pre- and postsurveys further indicate that novice tutors improve more, compared to veteran tutors, in their self-perceived oral/written communication skill levels. Qualitative findings from postmentorship interviews explain findings suggested by quantitative analysis, with contextual factors. This research study has bifold significance: "a theoretical perspective" on writing center work and research-supported professional development strategies. The findings of this study provide more food for thought on the subjects of how to design veteran–novice mentorships, how to target some transferable skills for professional development in the future, and how to exemplify the transferable skills in the survey to make those abstract constructs more concrete.

目的 期望获得含有全长hACE2基因序列的hACE2人源化小鼠模型,为心血管疾病、新冠肺炎发病机制研究、药物与疫苗研发提供重要工具.方法 将纯化含有hACE2完整编码序列的BAC质粒,利用小鼠受精卵显微注射和输卵管移植技术获得多个首建鼠,分别育种建系,通过杂交育种获得稳定遗传F2代小鼠品系,进一步对F2代小鼠hACE2基因整合、基因拷贝数、相对荧光定量PCR、Western Blot和免疫荧光进行分析.结果 育种获得hACE2-6-9、hACE2-14-3、hACE2-15-1和hACE2-15-2共4个小鼠品系.基因整合结果显示,hACE2-6-9、hACE2-15-1和hACE2-15-2小鼠品系含有完整的hACE2基因座位点及较长的基因调控区,hACE2-14-3含有完整的hACE2基因座位点及3'UTR较短的基因调控区.相对荧光定量PCR和Western Blot检测结果显示,hACE2-6-9、hACE2-15-1和hACE2-15-2品系小鼠肠道中hACE2 mRNA和蛋白表达水平较低,而整合较短调控序列的hACE2-14-3小鼠品系则表达水平较高.免疫荧光染色结果显示hACE2-15-1品系小鼠肾小管和肺血管内皮细胞中均有hACE2表达.结论 获得了含有全长基因序列的hACE2人源化BAC转基因小鼠,保留完整的hACE2启动子及基因调控区,从而为心血管疾病、新冠肺炎发病机制研究、hACE2基因表达调控机制研究和药物、疫苗的研发提供重要工具.

Background : There are remarkable genetic differences between animal major histocompatibility complex（MHC） systems and the human leukocyte antigen（HLA） system. HLA transgenic humanized mouse model systems offer a much better method to study the HLA-A-related principal mechanisms for vaccine development and HLA-Arestricted responses against infection in human. Methods : A recombinant gene encoding the chimeric HLA-A30 monochain was constructed. This HHD molecule contains the following: α1-α2 domains of HLA-A30, α3 and cytoplasmic domains of H-2D b , linked at its N-terminus to the C-terminus of human β2m by a 15-amino-acid peptide linker. The recombinant gene encoding the chimeric HLA-A30 monochain cassette was introduced into bacterial artificial chromosome（BAC） CH502-67J3 containing the HLA-A01 gene locus by Red-mediated homologous recombination. Modified BAC CH502-67J3 was microinjected into the pronuclei of wild-type mouse oocytes. This humanized mouse model was further used to assess the immune responses against influenza A virus（H1N1） pdm09 clinically isolated from human patients. Immune cell population, cytokine production, and histopathology in the lung were analyzed. Results : We describe a novel human β2m-HLA-A30（α1α2）-H-2D b （α3 transmembrane cytoplasmic）（HHD） monochain transgenic mouse strain, which contains the intact HLA-A01 gene locus including 49 kb 5’-UTR and 74 kb 3’-UTR of HLA-A01*01. Five transgenic lines integrated into the large genomic region of HLA-A gene locus were obtained, and the robust expression of exogenous transgene was detected in various tissues from A30-18# and A30-19# lines encompassing the intact flanking sequences. Flow cytometry revealed that the introduction of a large genomic region in HLA-A gene locus can influence the immune cell constitution in humanized mice. Pdm09 infection caused a similar immune response among HLA-A30 Tg humanized mice and wild-type mice, and induced the rapid increase of cytokines, including IFN-γ, TNF-α, and IL-6, in both HLA-A30 humanized Tg mice and wild-type mice. The expression of HLA-A30 transgene was dramatically promoted in tissues from A30-9# line at 3 days post-infection（dpi）. Conclusions : We established a promising preclinical research animal model of HLA-A30 Tg humanized mouse, which could accelerate the identification of novel HLA-A30-restricted epitopes and vaccine development, and support the study of HLA-A-restricted responses against infection in humans.

Transient receptor potential vanillic acid 2 (TRPV2) are well recognized for their contributions to neuronal development, cardiac function, immunity and cancer. However, the precise roles for this thermo TRPchannels in neurological disorder remain unknown. In this study, we employed the CRISPR/Cas9 system to generate genetic mutations of TRPV2. Genetic mutation of TRPV2 resulted in autistic-like phenotypes in mice accompanied with the disordered electrical signals recorded by multi-channels in vivo. To determine possible molecular mechanisms, western blotting was further used to assess the possible involvement of several autism-related proteins. The significantly decreased expression of the R2 subunit of the GABA-B receptor in the hippocampus was observed. Together, our findings suggest that genetic mutation of TRPV2 induces autism-like behavior, results in decreased expression of the R2 subunit of the GABA-B receptor.