Alzheimer's disease (AD) arises from pathological interactions among diverse brain cell types, but cell-specific proteomic changes remain underexplored. Here, we present deep proteomic profiling of sorted or proximity-labeled brain cells from AD mouse models (5xFAD and AppNL-G-F) at multiple ages, quantifying 13,411 proteins in microglia (three subtypes), astrocytes, oligodendrocyte precursor cells, and neurons. We identified 3,028 differentially abundant proteins across these cell types, the majority of which were not detected in bulk proteomic datasets, and constructed cell type-specific networks to define functional modules and hub proteins. Comparison with transcriptomic data revealed that ~30% of proteomic changes are RNA-independent. Further analyses uncovered cross-cell type signaling proteins conserved in human AD brains, such as pleiotrophin (Ptn), which is transcriptionally enriched in astrocytes but accumulates in microglia. Importantly, recombinant PTN directly activates induced microglia-like (iMG) human cells. Thus, these findings provide a comprehensive cell type-resolved proteomic atlas of AD models, highlighting novel intra- and intercellular signaling events.

Increased inflammation has been linked to behavioral pathogenesis in depression. Previous studies have shown that administration of inflammatory stimuli induces motivational deficits associated with reduced activation of the ventral striatum in association with reduced dopamine (DA) availability and release. However, the underlying mechanisms of inflammation-induced DA dysfunction remain largely unknown. Here, we investigated the in vitro effects of the inflammatory cytokine interleukin (IL)-6 on female and male human induced pluripotent stem cell (iPSC)-derived DAergic neurons from healthy volunteers. We identified inhibitory effects of IL-6 on female DA neurons, including reduced DA release, neuronal firing, velocity of synaptic vesicle (SV) transport, and density of docked SV, which was further supported by transcriptomic analyses. In contrast, male DA neurons exhibited an IL-6-induced compensatory phenotype, including increased velocity and density of SV and increased presynaptic terminal density. The long noncoding RNA (lncRNA) MIAT mediated these differences in male DA neurons, potentially via interaction with genes involved in the IL-6 signaling pathway and DA regulation. Moreover, by knocking out MIAT expression in male DA neurons, IL-6-induced deficits emerged, including reduced DA release, neuronal firing, and SV docking. Lastly, we found that the Janus kinase (JAK) inhibitor baricitinib reversed the inhibitory effects of IL-6 on female DA neurons. This work extends our understanding of the impact of inflammation on DA neurons, while identifying important sex differences and pharmacologic targets, ultimately laying the foundation for anti-inflammatory treatments of depressed patients with increased inflammation.

Drug repurposing offers a potential strategy to expand treatment options for conditions with limited therapies, but advancing repurposing candidates toward clinical implementation remains a challenge. Large-scale data, together with advanced genetic and epidemiological methods, may help address this gap. Here, we present an integrative digital medicine approach that combines genetically predicted transcriptomic signatures and perturbation screening for candidate identification with multi-cohort real-world validation for systematic evaluation of prioritized candidates. We applied this approach to Alzheimer's disease (AD), a disease with substantial unmet clinical need and persistent difficulty in developing effective therapies. We constructed AD signatures from genetically predicted expression changes across bulk tissues and microglia, then queried Connectivity Map profiles to identify compounds predicted to oppose these signatures. Aspirin emerged as a reproducible candidate across multiple signatures and underwent further evaluation. We then examined its association with incident AD in longitudinal electronic health record data from Vanderbilt University Medical Center and the NIH All of Us Research Program, as well as national insurance claims data. Across independent cohorts, aspirin initiation before age 65 was consistently associated with lower risk of incident AD, with signals suggesting that cumulative exposure and APOE ε4 status may influence effect size. Transcriptomic analysis of human cortical organoids provided additional experimental support, showing that aspirin more strongly opposed AD-related neuronal pathway alterations in wild-type organoids than in an organoid model of AD. This integrative approach offers a scalable strategy for genetically informed drug repurposing that bridges candidate discovery and clinical evaluation.

Introduction Hepatocellular carcinoma (HCC) is a leading cause of cancer-related mortality worldwide. Growing evidence indicates that the gut–liver axis plays a critical role in HCC pathogenesis through interactions between gut microbiota and liver physiology, involving microbial translocation, metabolite signaling, and immune and metabolic reprogramming within the hepatic microenvironment. Owing to its multi-component and multi-target properties, Traditional Chinese Medicine (TCM) represents a promising strategy for modulating this axis. This review summarizes current evidence on gut microbial communities, key metabolites, and the microbiota-modulating potential of TCM in HCC progression. Methods This review examines current evidence on gut microbiota–mediated mechanisms in HCC and the modulatory effects of TCM via the gut–liver axis. Literature was retrieved from PubMed, Web of Science, Embase, CNKI up to December 2025. Eligible studies included experimental, animal, and clinical investigations addressing gut microbiota dysbiosis, microbial metabolites, intestinal barrier dysfunction, intratumoral microbiota, and TCM interventions in HCC or related chronic liver diseases. Results Gut microbiota dysbiosis is a key driver of HCC through disruption of the gut–liver axis, characterized by reduced microbial diversity, intestinal barrier impairment, and microbial translocation–induced inflammation. Microbiota-derived metabolites, including lipopolysaccharides, secondary bile acids, short-chain fatty acids, ethanol metabolites, amino acid derivatives and lipid metabolites play central roles in regulating inflammatory signaling, immune responses, and tumor metabolism. TCM formulas and active compounds can restore gut microbial balance, strengthen intestinal barrier function, regulate microbial metabolites, and suppress pro-inflammatory and pro-tumorigenic signaling pathways, thereby attenuating hepatic inflammation and inhibiting HCC progression. Discussion Gut microbiota dysbiosis along the gut–liver axis plays a central role in HCC representing a promising therapeutic target. TCM may modulate this axis through multi-component, multi-target mechanisms, but further clinical and multi-omics studies are required to establish causal links and enable precision therapy.

Fragile X-associated tremor/ataxia syndrome (FXTAS) is a neurodegenerative disorder caused by CGG repeat expansions in the FMR1 gene. While CGG repeat toxicity is established, the precise molecular mechanisms driving neurodegeneration remain unclear. Here, we show that a multi-omics strategy combined with TWAS reveals brain-region-specific molecular signatures and striking gene dysregulation in inhibitory neurons. Using conditional mouse models, we demonstrate that selective expression of expanded CGG repeats in GABAergic neurons is sufficient to recapitulate key pathologic hallmarks of FXTAS. We identify PRKCG as a genetic modifier of FXTAS, with cross-species evidence linking its overexpression to disease onset. Many dysregulated mRNAs in GABAergic neurons are targets of hnRNPA2/B1, an RNA-binding protein sequestered by CGG repeat RNA. Functional screening in Drosophila further establishes PRKCG as a potent modulator of CGG-associated neurotoxicity. These findings uncover a critical role of GABAergic neurons in FXTAS pathogenesis and position PRKCG as a promising therapeutic target.

The basolateral amygdala (BLA) is a key structure for processing threat and emotional information, and plays a key role in controlling the fear memory. Previous research has suggested that the extinction procedure generates a new memory that coexists with the original fear memory, and that interneurons play an important role in this process. However, the mechanisms that control the competing extinction and fear memories in BLA are not yet fully understood. To investigate these mechanisms, we developed a chemogenetic strategy that offers improved sensitivity and specificity for tagging neurons during defined behavioral epochs. This was achieved by genetically targeting GABAergic interneurons for activity-dependent manipulation, allowing us to directly probe the functional role of inhibitory ensembles recruited during fear extinction. We found that silencing extinction-tagged BLA neurons or stimulating fear acquisition-tagged BLA neurons, led to a relapse of fear memory. Additionally, we observed that silencing BLA GABAergic neurons, or more specifically, silencing the extinction-tagged BLA GABAergic neurons, restored fear expression after extinction, while inhibiting acquisition-tagged BLA GABAergic neurons did not impair fear memory retrieval. Our results indicate that specific inhibitory GABAergic BLA engrams are established during fear extinction, which interfere with existing fear memory-related neural circuits and suppress conditioned fear memory. These findings provide insights into the neural mechanisms underlying fear extinction and suggest that BLA GABAergic neurons are potential targets for therapeutic interventions in cognitive disorders such as posttraumatic stress disorder (PTSD).

The convergence of oncology and neurodevelopmental research has uncovered compelling links between genetic factors and mental health. Among these, B-cell chronic lymphocytic leukemia/lymphoma 9 (BCL9), located on human chromosome 1q21.1, has been identified as a key oncogene in precursor B-cell acute lymphoblastic leukemia (B-ALL). Subsequent studies have established that Bcl9 promotes tumorigenesis through activation of the Wnt/β-catenin signaling pathway. Emerging evidence from human genetic studies further implicates common variants of Bcl9 in increased susceptibility to neurodevelopmental disorders, including schizophrenia and bipolar disorder. Despite growing interest in its role beyond cancer, the neurobiological functions of Bcl9 remain poorly defined. In this study, we systematically investigated the role of Bcl9 in neurodevelopment using mouse models. By employing in utero electroporation-mediated gene knockdown techniques and conditional knockout (cKO) mice, we demonstrate that Bcl9 deficiency disrupts Wnt-dependent signal transduction and leads to upregulation of voltage-gated sodium channel (Nav) expression, thereby results in increased sodium current density but paradoxically reduced action potential firing. Based on our experimental findings, Bcl9 and/or the Bcl9 complexes may regulate the promoter region of the Scn1b gene and the area near the exons of Scn2a. Collectively, our findings establish Bcl9 as a critical regulator of neuronal excitability and migration, providing a mechanistic framework that links ion channel dysregulation to the pathophysiology of neurodevelopmental disorders.

BACKGROUND:Trauma-Induced Coagulopathy is a severe condition that rapidly manifests following traumatic injury and is characterized by shock, hypoperfusion, and vascular damage. This study employed bioinformatics methods to identify crucial hub genes and pathways associated with TIC. METHODS:Microarray datasets (accession number GSE223245) were obtained from the Gene Expression Omnibus (GEO) database. The data were subjected analyses to identify the Differentially Expressed Genes (DEGs), which were further subjected to GO and KEGG pathway analyses. Subsequently, a Protein-Protein Interaction (PPI) network was constructed and hub DEGs closely linked to TIC were identified using CytoHubba, MCODE, and CTD scores. The diagnostic value of these hub genes was evaluated using Receiver Operating Characteristic (ROC) analysis. RESULTS:Among the analyzed genes, 269 were identified as DEGs, comprising 103 upregulated and 739 downregulated genes. Notably, several significant hub genes were associated with the development of TIC, as revealed by bioinformatic analyses. CONCLUSIONS:This study highlights the critical impact of newly discovered genes on the development and progression of TIC. Further validation through experimental research and clinical trials is required to confirm these findings.

Many emerging molecular targets are implicated in AD, prompting the need for models to efficiently and rapidly evaluate their functional importance. Development of human induced pluripotent stem cell (hiPSC) models has accelerated our ability to study these processes across AD-relevant cell types. Advantages of hiPSC cultures include their ability to regenerate, receive disease-specific alterations in isogenic backgrounds, and withstand experimental variations in a controlled setting. Conversely, stringent culture conditions and prolonged differentiation limit the utility of hiPSC-derived models, especially in high-density plates. Leveraging our expertise at the Emory-Sage-SGC-Jax TREAT-AD Center in developing hiPSC-derived neuron cultures and miniaturization technologies for high-throughput screening (HTS), we optimized the hiPSC-derived neuron long-term culture into a 384-well HTS format. A panel of functional HTS and image-based high-content screening (HCS) assays were developed to rapidly evaluate many experimental variables, including target modulations and small molecule treatment. The hiPSC-derived neuron cells were seeded into the 384-well PDL-coated plate and maintained in neuron maturation culture medium for 4 weeks. The medium was changed twice weekly before compound treatment. The cells were proceeded to various HTS/HCS assays, including live-cell based, such as mitochondrial function, cell viability, oxidative stress; and/or fixed-cell based, such as immunofluorescence (IF) staining for neuron morphology and synaptic The optimization of hiPSC-derived neuron culture conditions resulted in healthy, long-term differentiated neurons in a 384-well HTS format. Copious data across multiple phenotypes were obtained through a panel of multiplexed functional assays for matured neurons. The optimization of HTS/HCS functional assays led to robust assay performance, which allows the sensitive detection of phenotypic changes with small molecules and biological perturbations of the neurons. We optimized the long-term hiPSC-derived neuron culture into a 384-well HTS format for modeling AD and developed a panel of functional HTS/HCS assays for screening small molecules and biological perturbagens. The results from screening a set of well-annotated small molecules with known biological targets and activities will enable us to rapidly advance knowledge of AD targets in hiPSC-derived, AD-relevant cell types. Our HTS neuron long-term culture system will make expanded screening with large compound libraries feasible to accelerate AD drug discovery.

With the sustained growth of the economy and significant changes in social demographics, the issue of elderly-related diseases has increasingly drawn attention, particularly. Alzheimer’s disease (AD), as a representative disease of neurodegenerative diseases, has become a major challenge, affecting the health and quality of life of the elderly population severely. In recent years, the incidence, prevalence and mortality rates of AD have increased in China, imposing substantial economic burdens on families, society and the entire healthcare system. To proactively address this challenge and respond to the national ‘Healthy China Action’ initiative, leading experts from authoritative institutions jointly authored the China Alzheimer Report 2025. Building on previous editions, this report updates epidemiological data on AD in China, thoroughly analyses the latest economic burdens of the disease and comprehensively evaluates the current status of AD diagnosis and treatment services, as well as the allocation of public health resources in our country. Its release reflects China’s progress in AD research and prevention, underscores societal concern for elderly health and aims to provide scientific guidance and data support for AD prevention, diagnosis and treatment. It also facilitates academic exchanges and cooperation, enhancing public awareness and promoting active participation in elderly healthcare, towards achieving ‘healthy ageing’ in China.

MicroRNAs (miRNAs) and long non-coding RNAs (lncRNAs) regulate broad gene networks through distinct mechanisms, which govern normal brain development and function but are dysregulated in schizophrenia (SCZ). However, how disease-risk miRNAs and lncRNAs co-operate to form pathogenic pathways in SCZ brains remain poorly understood. In this study, we identified a novel miRNA-lncRNA pathway in which the well-recognized SCZ-risk factor miR-137 enhances expression of the SCZ-risk lncRNA GOMAFU in human neuron development. We found significant up-regulation of GOMAFU during differentiation of multiple types of human neurons in vivo and in culture. Interestingly, the accumulation of histone acetylation, which activates numerous neuronal genes, down-regulates GOMAFU in iPSC-derived human neurons through inducing transcription repressors of GOMAFU, represented by the miR-137-target E2F6. We further demonstrated that miR-137 is necessary and sufficient for enhancing GOMAFU expression in a human neuronal progenitor cell (NPC) line and observed co-regulation of MIR137 with GOMAFU during normal human neuronal development and in SCZ brains. Moreover, we identified human NPC transcriptomic changes induced by miR-137 and discovered that miR-137 integrates functional co-operation of histone acetylation and transcription factors to promote GOMAFU expression. Notably, a significant number of miR-137-regulated transcription factors are predicted to bind the GOMAFU promoter and affected in SCZ brains, forming a highly interactive molecular network. Together, these results unveil the SCZ risk miR-137-GOMAFU non-coding RNA pathway connected by SCZ-affected transcription factors, providing a new mode of functional integration of non-coding and coding risk genes of SCZ that contributes to the complex etiology.

Neuroinflammation is a key driver of Alzheimer's disease and an emerging therapeutic target. The p38/MK2 pathway regulates microglial cytokine production, yet previous attempts have not yielded modulators with clinically suitable properties. Here, we apply an integrative structure-guided and screening strategy to identify small-molecule disruptors of the p38/MK2 protein-protein interaction (PPI). Virtual screening of FDA-approved drugs prioritized nilotinib, a BCR-ABL inhibitor, as a putative PPI disruptor. Biochemical and molecular dynamics analyses confirmed that nilotinib binds to p38, blocks MK2 association, and suppresses cytokine release in microglia. Guided by these findings, we developed a lysate-based TR-FRET ultrahigh-throughput assay that identified additional inhibitors, including α1-adrenergic antagonists doxazosin, terazosin, and alfuzosin. These compounds suppressed cytokine induction via docking groove blockade. Together, these results establish a non-ATP-competitive approach for selectively targeting the p38/MK2 complex and highlight the translational potential of drug repurposing to modulate neuroinflammation in Alzheimer's disease.

Epidermal growth factor receptor inhibitors (EGFRis) are used to treat many cancers, but their use is complicated by the development of a skin rash that may be severe, limiting their use and adversely affecting patient quality of life. Most studies of EGFRi-induced rash have focused on the fully developed stage of this skin disorder, and early pathological changes remain unclear. We analyzed high-throughput transcriptome sequencing of skin samples from rats exposed to the EGFRi afatinib and identified that keratinocyte activation is an early pathological alteration in EGFRi-induced rash. Mechanistically, the induction of S100 calcium-binding protein A9 (S100A9) occurred before skin barrier disruption and led to keratinocyte activation, resulting in expression of specific cytokines, chemokines, and surface molecules such as interleukin 6 (Il6) and C-C motif chemokine ligand 2 (CCL2) to recruit and activate monocytes through activation of the Janus kinase (JAK)–signal transducers and activators of transcription (STAT) pathway, further recruiting more immune cells. Topical JAK inhibition suppressed the recruitment of immune cells and ameliorated the severity of skin rash in afatinib-treated rats and mice with epidermal deletion of EGFR, while having no effect on EGFRi efficacy in tumor-bearing mice. In a pilot clinical trial (NCT05120362), 11 patients with EGFRi-induced rash were treated with delgocitinib ointment, resulting in improvement in rash severity by at least one grade in 10 of them according to the MASCC EGFR inhibitor skin toxicity tool (MESTT) criteria. These findings provide a better understanding of the early pathophysiology of EGFRi-induced rash and suggest a strategy to manage this condition.

TIMELESS (TIM) is a circadian gene which is implicated in the regulation of daily rhythm, DNA replication and repair, and cancer initiation and progression. Nevertheless, the role of TIM in endometrial cancer (EC) development is largely unknown. Bioinformatics analysis showed that TIM was aberrantly up-regulated in EC tissues and positively correlated with clinical or histological grade of EC. Functional studies showed that TIM knockdown reduced EC cell viability and restrained EC cell migration in vitro, as well as blocked xenograft tumor growth in vivo. Mechanistically, HMGB1 transcriptionally up-regulated TIM expression in EC cells. In addition, TIM could activate the transcription of the canonical Wnt ligand WNT8B, and TIM depletion could reduce the malignant potential of EC cells largely by targeting and down-regulating WNT8B. As a conclusion, HMGB1/TIM/WNT8B signal cascade was identified in this study for the first time. HMGB1 exerted its oncogenic role by activating the transcription of TIM, leading to the activation of Wnt signaling and EC progression.

BACKGROUND:Kindlin-3 in platelets plays an essential role in supporting integrin αIIbβ3 activation, platelet spreading, aggregation, and clot retraction by binding to the integrin β3 cytoplasmic tail. However, the mechanism by which kindlin-3 mediates the crosstalk between integrin αIIbβ3 and myosin in platelets remains unknown. OBJECTIVES:To examine the role of myosin light chain 6 (Myl6) in supporting integrin αIIbβ3 activation in platelets. METHODS:Myl6fl/flPF4-Cre mice with a deficiency of Myl6 in the megakaryocyte lineage were generated, and integrin αIIbβ3 activation in Myl6-deficient platelets was analyzed. RESULTS:We identified a novel kindlin-3 binding protein, Myl6, an essential light chain of myosin in platelets. Myl6fl/flPF4-Cre mice exhibited significant macrothrombocytopenia resulting from defective proplatelet formation. In the absence of Myl6, integrin αIIbβ3 activation in platelets was significantly suppressed, and platelet aggregation was substantially impaired. Interestingly, the deficiency of Myl6 in platelets preferentially affected the binding of a multivalent ligand compared to a monovalent ligand to integrin αIIbβ3 upon activation, indicating that Myl6 may contribute to the avidity modulation of integrin αIIbβ3 by binding to kindlin-3. Furthermore, blood coagulation ability was impaired in Myl6fl/flPF4-Cre mice, and consistently, these mice exhibited defects in both hemostatic and thrombotic functions. CONCLUSION:In summary, these results suggest that Myl6, as a novel kindlin-3 binding partner, is required to support integrin αIIbβ3 activation in platelets, which plays an important role in both hemostasis and thrombosis.

BackgroundKabuki syndrome (KS) is a rare developmental disorder characterised by multiple congenital anomalies and intellectual disability.UTX(ubiquitously transcribed tetratricopeptide repeat, X chromosome), which encodes a histone demethylase, is one of the two major pathogenic risk genes for KS. Although intellectual disability is a key phenotype of KS, the role ofUTXin cognitive function remains unclear. Currently, no targeted therapies are available for KS.AimsThis study aimed to investigate howUTXregulates cognition, to explore the mechanisms underlyingUTXdysfunction and to identify potential molecular targets for treatment.MethodsWe generatedUTXconditional knockout mice and found thatUTXdeletion downregulated calmodulin transcription by disrupting H3K27me3 (trimethylated histone H3 at lysine 27) demethylation.ResultsUTX-knockout mice showed decreased phosphorylation of calcium / calmodulin-dependent protein kinase II, impaired long-term potentiation and deficit in remote contextual fear memory. These effects were reversed by an Food and Drug Administration-approved drug desipramine.ConclusionsOur results reveal an epigenetic mechanism underlying the important role ofUTXin synaptic plasticity and cognitive function, and suggest that desipramine could be a potential treatment for KS.

RNA-binding proteins (RBPs) control messenger RNA fate in neurons. Here, we report a mechanism that the stimuli-induced neuronal translation is mediated by phosphorylation of a YTHDF1-binding protein FMRP. Mechanistically, YTHDF1 can condense with ribosomal proteins to promote the translation of its mRNA targets. FMRP regulates this process by sequestering YTHDF1 away from the ribosome; upon neuronal stimulation, FMRP becomes phosphorylated and releases YTHDF1 for translation upregulation. We show that a new small molecule inhibitor of YTHDF1 can reverse fragile X syndrome (FXS) developmental defects associated with FMRP deficiency in an organoid model. Our study thus reveals that FMRP and its phosphorylation are important regulators of activity-dependent translation during neuronal development and stimulation and identifies YTHDF1 as a potential therapeutic target for FXS in which developmental defects caused by FMRP depletion could be reversed through YTHDF1 inhibition.

Background The lower limb mechanical axis was used to assess the severity of knee osteoarthritis (KOA) with varus/valgus deformity and the accuracy of targeted lower limb alignment correction after operation by conventional X-rays. There are lots of parameters to assess the gait in elder patients such as velocity, stride length, step width and swing/stance ratio by knee joint movement analysis system. However, the correlation between the lower limb mechanical axis and gait parameters is not clear. This study is aimed at obtaining the accuracy of the lower limb mechanical axis by the knee joint movement analysis system and the correlation between the lower limb mechanical axis and gait parameters. Methods We analysed 3D knee kinematics during ground gait of 99 patients with KOA and 80 patients 6 months after the operations with the vivo infrared navigation 3D portable knee joint movement analysis system (Opti-Knee®, Innomotion Inc, Shanghai, China). The HKA (Hip-Knee-Ankle) value was calculated and compared to X-ray findings. Results HKA absolute variation after the operation was 0.83 ± 3.76°, which is lower than that before the operation (5.41 ± 6.20°, p = 0.001) and also lower than the entire cohort (3.36 ± 5.72). Throughout the cohort, a significant correlation with low coefficients (r = -0.19, p = 0.01) between HKA value and anterior-posterior displacement was found. In comparing the HKA values measured on the full-length alignment radiographs and 3D knee joint movement analysis system (Opti-Knee), there was a significant correlation with moderate to high coefficients (r = 0.784 to 0.976). The linear correlation analysis showed that there was a significant correlation between the values of HKA measured by X-ray and movement analysis system (R 2 = 0.90, p < 0.01). Conclusions Data with equivalent results as HKA, the 6DOF of the knee and ground gait data could be provided by infrared navigation based 3D portable knee joint movement analysis system comparing with the conventional X-rays. There is no significant effect of HKA on the kinematics of the partial knee joint.

Brain organoids represent a new model system for studying developmental human neurophysiology. Methods for studying the electrophysiology and morphology of single neurons in organoids require acute slices or dissociated cultures. While these methods have advantages (e.g., visual access, ease of experimentation), they risk damaging cells and circuits present in the intact organoid. To access single cells within intact organoid circuits, we have demonstrated a method for fixturing and performing whole cell patch clamp recording from intact brain organoids using both manual and automated tools. We demonstrate applied electrophysiology methods development followed by an integration of electrophysiology with reconstructing the morphology of the neurons within the brain organoid using dye filling and tissue clearing. We found that whole cell patch clamp recordings could be achieved both on the surface and within the interior of intact human brain organoids using both manual and automated methods. Manual experiments were higher yield (53 % whole cell success rate manual, 9 % whole cell success rate automated), but automated experiments were more efficient (30 patch attempts per day automated, 10 patch attempts per day manual). Using these methods, we performed an unbiased survey of cells within human brain organoids between 90 and 120 days in vitro (DIV) and present preliminary data on morphological and electrical diversity in human brain organoids. The further development of intact brain organoid patch clamp methods could be broadly applicable to studies of cellular, synaptic, and circuit-level function in the developing human brain.

OBJECTIVE AND DESIGN:An accumulating body of evidence has shown that gut microbiota is involved in regulating inflammation; however, it remains undetermined if and how gut microbiota plays an important role in modulating deep venous thrombosis (DVT), which is an inflammation-involved thrombotic event.SUBJECTS:Mice under different treatments were used in this study.METHODS AND TREATMENT:We induced stenosis DVT in mice by partially ligating the inferior vena cava. Mice were treated with antibiotics, prebiotics, probiotics, or inflammatory reagents to modulate inflammatory states, and their effects on the levels of circulating LPS and DVT were examined.RESULTS:Antibiotic-treated mice or germ-free mice exhibited compromised DVT. Treatment of mice with either prebiotics or probiotics effectively suppressed DVT, which was accompanied with the downregulation of circulating LPS. Restoration of circulating LPS in these mice with a low dose of LPS was able to restore DVT. LPS-induced DVT was blocked by a TLR4 antagonist. By performing proteomic analysis, we identified TSP1 as one of the downstream effectors of circulating LPS in DVT.CONCLUSION:These results suggest that gut microbiota may play a nonnegligible role in modulating DVT by leveraging the levels of LPS in circulation, thus shedding light on the development of gut microbiota-based strategies for preventing and treating DVT.