Osteoarthritis (OA), a highly prevalent degenerative joint disease worldwide, is defined by articular cartilage degradation, abnormal bone remodeling, and persistent chronic inflammation. It severely compromises patients’ quality of life, and currently, there is no radical cure. Abnormal mechanical stress is widely regarded as a core driver of OA pathogenesis, and the exploration of mechanical signal perception and transduction mechanisms has become crucial for deciphering OA’s pathophysiological processes. Piezo1, a key mechanosensitive cation channel belonging to the Piezo protein family, has recently gained significant attention due to its pivotal role in mediating cellular responses to mechanical stimuli in joint tissues. This review systematically examines Piezo1’s expression patterns, regulatory mechanisms, and pathological functions in OA, with a particular focus on its dual roles in modulating chondrocyte homeostasis and bone metabolism disorders, while also delving into the underlying molecular signaling pathways and potential therapeutic implications. Piezo1, consisting of approximately 2 500 amino acids and forming a unique trimeric propeller-like structure, is widely expressed in chondrocytes, osteocytes, mesenchymal stem cells, and synovial cells. It exhibits permeability to cations such as Ca²⁺, K⁺, and Na⁺, and directly responds to membrane tension changes induced by mechanical stimuli like fluid shear stress and mechanical overload. In OA patients and animal models, Piezo1 expression is significantly upregulated, especially in cartilage regions subjected to abnormal mechanical stress (e.g., human temporomandibular joint cartilage). This overexpression is closely associated with aggravated cartilage degeneration, increased chondrocyte apoptosis, accelerated cellular senescence, and intensified inflammatory responses. Mechanical overload and pro-inflammatory cytokines (e.g., IL-1β) are key inducers of Piezo1 upregulation: IL-1β activates the PI3K/AKT/mTOR signaling pathway to enhance Piezo1 expression, forming a pathogenic positive feedback loop that inhibits chondrocyte autophagy, promotes apoptosis, and further accelerates joint degeneration. Mechanistically, Piezo1 mediates OA progression through multiple interconnected pathways. When activated by mechanical stress, Piezo1 triggers excessive Ca²⁺ influx, leading to endoplasmic reticulum stress (ERS) and mitochondrial dysfunction, which directly induce chondrocyte apoptosis. This process involves the activation of downstream signaling cascades such as cGAS-STING and YAP-MMP13/ADAMTS5. YAP, a transcriptional regulator, upregulates the expression of matrix metalloproteinase 13 (MMP13) and aggrecanase (ADAMTS5), thereby accelerating cartilage matrix degradation. Additionally, Piezo1-driven Ca²⁺ overload promotes the accumulation of reactive oxygen species (ROS) and upregulates senescence markers (p16 and p21), accelerating chondrocyte senescence via the p38MAPK and NF-κB pathways. Senescent chondrocytes secrete senescence-associated secretory phenotype (SASP) factors (e.g., IL-6, IL-1β), further amplifying joint inflammation. In terms of bone metabolism, Piezo1 maintains joint homeostasis by promoting the differentiation of fibrocartilage stem cells into chondrocytes and balancing bone formation and resorption through regulating the FoxC1/YAP axis and RANKL/OPG ratio. Therapeutically, targeting Piezo1 shows promising potential. Preclinical studies have demonstrated that Piezo1 inhibitors (e.g., GsMTx4) can reduce joint damage and alleviate pain in OA mice. Simultaneously, siRNA-mediated co-silencing of Piezo1 and TRPV4 (another mechanosensitive channel) decreases intracellular Ca²⁺ concentration, inhibits chondrocyte apoptosis, and promotes cartilage repair. Conditional knockout of Piezo1 using Gdf5-Cre transgenic mice alleviates cartilage degeneration in post-traumatic OA models by downregulating MMP13 and ADAMTS5 expression. Despite existing challenges, such as off-target effects of inhibitors, inefficient local drug delivery, and interindividual genetic variability, strategies like developing selective Piezo1 antagonists, optimizing targeted nanocarriers, and combining Piezo1-targeted therapy with physical therapy provide viable avenues for clinical translation. The authors propose that Piezo1 serves as a critical therapeutic target for OA, and future research should focus on deciphering its context-dependent regulatory networks, developing tissue-specific intervention strategies, and validating their efficacy and safety in clinical trials to address the unmet medical needs of OA patients.

With the rapid development of the biopharmaceutical field, the efficient and simultaneous extraction of multiple biological components from biological samples has become a critical process for advancing scientific research. The ability to simultaneously extract various molecular components such as metabolites, DNA, RNA, and proteins is pivotal for multi-omics studies, which aim to comprehensively understand the molecular mechanisms of biological systems. Traditional methods often extract these components separately, leading to challenges such as sample loss, time consumption, contamination, and inconsistencies across different data types. In contrast, simultaneous extraction techniques address these issues by maintaining the consistency of each biological component’s physiological state, improving data reliability and facilitating integration across omic platforms. This review systematically summarizes recent advances in simultaneous extraction technologies, focusing on methods such as methanol/chloroform extraction, TRIzol reagent extraction, and modified Folch extraction, which have shown significant promise in improving the efficiency and integrity of biological sample preparation. These methods offer various advantages, such as reduced sample volume requirements, decreased contamination risk, and enhanced extraction consistency, which are crucial for studies involving small sample sizes or precious clinical specimens. Among these, methanol/chloroform extraction stands out for its simplicity, low cost, and ability to extract a wide range of biological molecules. However, it does face limitations, such as its inefficiency in extracting lipids and potential RNA contamination. On the other hand, the TRIzol reagent method has become a widely adopted technique due to its ability to simultaneously isolate RNA, proteins, and metabolites from the same sample. Despite its effectiveness, the TRIzol method has limitations in RNA quality, especially when handling complex samples or those with high protein content. Modified Folch extraction, which combines liquid-liquid extraction with commercial kits, offers a highly efficient way to extract polar metabolites, lipids, RNA, DNA, and proteins from small tissue samples. This method has proven advantageous in terms of extraction yield, especially for challenging or rare samples, although it requires precise handling to avoid cross-contamination between phases. The integration of automated platforms, microfluidics, and high-throughput systems is another exciting avenue for improving simultaneous extraction. Automation facilitates large-scale, reproducible sample processing with minimal human error, while microfluidics provides high precision in sample handling and enables real-time monitoring of extraction efficiency. These innovations not only enhance the speed and reproducibility of sample preparation but also open new possibilities for single-cell analysis, where sample volumes are often limited, and extraction efficiency is critical. In addition to the technical aspects, the review also highlights the importance of optimizing extraction protocols for specific sample types, such as clinical tissues, plants, and microorganisms. For example, the challenge of extracting multiple components from cancer tissues, where sample degradation and contamination risks are high, can be mitigated by carefully selecting extraction reagents and minimizing sample handling steps. Similarly, in plant studies, where metabolite diversity is vast, the simultaneous extraction methods must be optimized to account for the unique composition of plant tissues, which often include complex secondary metabolites and cell wall components. Looking forward, the development of more efficient and standardized simultaneous extraction methods will be crucial for advancing multi-omics research. There is a growing need for protocols that can be tailored to specific research needs, ensuring both reproducibility and flexibility in diverse applications. Additionally, combining these extraction methods with high-resolution analytical techniques such as mass spectrometry and next-generation sequencing will further enhance the potential of multi-omics studies to provide comprehensive insights into biological systems. As these technologies continue to evolve, their application in personalized medicine, environmental research, and agriculture holds great promise for addressing critical scientific challenges. In conclusion, while simultaneous extraction technologies have made significant strides, several challenges remain in optimizing extraction efficiency, ensuring reproducibility, and reducing costs. Future research should focus on refining extraction protocols, developing innovative extraction reagents, and expanding the scope of these methods to cater to a broader range of biological samples. Ultimately, the continued integration of these advanced techniques will revolutionize the way biological samples are prepared, analyzed, and understood in the context of multi-omics research.

With the rapid development of the biopharmaceutical field, the efficient and simultaneous extraction of multiple biological components from biological samples has become a critical process for advancing scientific research. The ability to simultaneously extract various molecular components such as metabolites, DNA, RNA, and proteins is pivotal for multi-omics studies, which aim to comprehensively understand the molecular mechanisms of biological systems. Traditional methods often extract these components separately, leading to challenges such as sample loss, time consumption, contamination, and inconsistencies across different data types. In contrast, simultaneous extraction techniques address these issues by maintaining the consistency of each biological component’s physiological state, improving data reliability and facilitating integration across omic platforms. This review systematically summarizes recent advances in simultaneous extraction technologies, focusing on methods such as methanol/chloroform extraction, TRIzol reagent extraction, and modified Folch extraction, which have shown significant promise in improving the efficiency and integrity of biological sample preparation. These methods offer various advantages, such as reduced sample volume requirements, decreased contamination risk, and enhanced extraction consistency, which are crucial for studies involving small sample sizes or precious clinical specimens. Among these, methanol/chloroform extraction stands out for its simplicity, low cost, and ability to extract a wide range of biological molecules. However, it does face limitations, such as its inefficiency in extracting lipids and potential RNA contamination. On the other hand, the TRIzol reagent method has become a widely adopted technique due to its ability to simultaneously isolate RNA, proteins, and metabolites from the same sample. Despite its effectiveness, the TRIzol method has limitations in RNA quality, especially when handling complex samples or those with high protein content. Modified Folch extraction, which combines liquid-liquid extraction with commercial kits, offers a highly efficient way to extract polar metabolites, lipids, RNA, DNA, and proteins from small tissue samples. This method has proven advantageous in terms of extraction yield, especially for challenging or rare samples, although it requires precise handling to avoid cross-contamination between phases. The integration of automated platforms, microfluidics, and high-throughput systems is another exciting avenue for improving simultaneous extraction. Automation facilitates large-scale, reproducible sample processing with minimal human error, while microfluidics provides high precision in sample handling and enables real-time monitoring of extraction efficiency. These innovations not only enhance the speed and reproducibility of sample preparation but also open new possibilities for single-cell analysis, where sample volumes are often limited, and extraction efficiency is critical. In addition to the technical aspects, the review also highlights the importance of optimizing extraction protocols for specific sample types, such as clinical tissues, plants, and microorganisms. For example, the challenge of extracting multiple components from cancer tissues, where sample degradation and contamination risks are high, can be mitigated by carefully selecting extraction reagents and minimizing sample handling steps. Similarly, in plant studies, where metabolite diversity is vast, the simultaneous extraction methods must be optimized to account for the unique composition of plant tissues, which often include complex secondary metabolites and cell wall components. Looking forward, the development of more efficient and standardized simultaneous extraction methods will be crucial for advancing multi-omics research. There is a growing need for protocols that can be tailored to specific research needs, ensuring both reproducibility and flexibility in diverse applications. Additionally, combining these extraction methods with high-resolution analytical techniques such as mass spectrometry and next-generation sequencing will further enhance the potential of multi-omics studies to provide comprehensive insights into biological systems. As these technologies continue to evolve, their application in personalized medicine, environmental research, and agriculture holds great promise for addressing critical scientific challenges. In conclusion, while simultaneous extraction technologies have made significant strides, several challenges remain in optimizing extraction efficiency, ensuring reproducibility, and reducing costs. Future research should focus on refining extraction protocols, developing innovative extraction reagents, and expanding the scope of these methods to cater to a broader range of biological samples. Ultimately, the continued integration of these advanced techniques will revolutionize the way biological samples are prepared, analyzed, and understood in the context of multi-omics research.

Objectives: . This study aimed to investigate the regulatory functions of gamma-aminobutyric acid (GABA)ergic neural circuits from the inferior colliculus (IC) to the medial geniculate body (MGB) in salicylate-induced tinnitus. Methods: . Mice were treated with salicylate to induce tinnitus, and tinnitus-like behaviors were evaluated via gap prepulse inhibition of acoustic startle. Using combined viral tracing methodologies, we identified and mapped the pathways and connections from the IC to the MGB. Furthermore, we employed Gq-coupled human M3 designer receptors exclusively activated by designer drugs (DREADDs) and Gi-coupled human M4 DREADDs to achieve targeted excitation or suppression of GABAergic neurons in the IC and MGB. Following the administration of clozapine N-oxide, which binds to these receptors, we modulated these neural circuits to assess their impact on tinnitus severity in a mouse model. Results: . Our findings demonstrated that mice exposed to salicylate exhibited tinnitus-like behaviors. GABAergic neurons projecting retrogradely from the MGB to the IC were primarily concentrated in the external nucleus of the IC. After clozapine N-oxide administration, chemogenetic activation of IC-MGB GABAergic neurons aggravated salicylate-induced tinnitus. Additionally, activation of GABAergic neurons between the IC and MGB induced the perception of tinnitus even without salicylate. However, chemogenetic inhibition of the IC-MGB GABAergic circuit did not reverse salicylate-induced tinnitus. Conclusion . These findings suggest that activation of the IC-MGB GABAergic neural circuit may contribute to tinnitus generation through a mechanism distinct from that of salicylate-induced tinnitus. This study provides novel insights into the mechanisms underlying tinnitus.

Multidrug-resistant (MDR) bacterial infections have emerged as a serious challenge of global public health crisis. The overuse and misuse of conventional antibiotics have dramatically accelerated the emergence, evolution and worldwide spread of drug-resistant bacterial strains, necessitating urgent exploration of novel antibacterial strategies. Bacteriophages serve as natural bacterial predators offering distinct advantages including high host specificity, autonomous self-replication capabilities and cost-effective large-scale production. However, wild-type phages present significant clinical limitations due to their narrow host ranges, susceptibility to rapid immune clearance and poor penetration of bacterial biofilms, which severely restrict their therapeutic applications. The convergence of synthetic biology, nanotechnology and advanced gene editing technologies has accelerated the development of engineered bacteriophage platforms, providing programmable, scalable and clinically translatable pathways to overcome these inherent biological constraints. Here, we systematically delineate four fundamental strategies for engineered bacteriophage development. Chemical modification utilizes reactive functional groups such as amino, carboxyl and thiol moieties on capsid proteins through esterification, amidation or click chemistry reactions to achieve precise drug conjugation and surface functionalization. In vivo editing encompasses ultraviolet or chemical mutagenesis for random mutation induction, homologous recombination for targeted genetic alterations, recombineering methodologies including electroporation-mediated bacteriophage recombination engineering, and CRISPR-Cas systems for precise genome editing to enable exact genetic reconstruction and host range reprogramming. In vitro synthesis leverages genome engineering platforms where intact phage genomes are transferred into yeast or host bacteria to facilitate highly efficient homologous recombination, enabling large DNA fragment assembly and cross-gene host range expansion without bacterial toxicity constraints. Directed evolution combines artificial selection through mutation library screening with rational design approaches involving chimeric receptor binding protein construction or site-specific mutagenesis, effectively balancing the discovery of unknown adaptive pathways with targeted host specificity modification. Moreover, we comprehensively discuss therapeutic applications across diverse clinical scenarios. Engineered bacteriophage effectively disrupt bacterial biofilms through sophisticated functionalized delivery platforms including nanozyme-conjugated phages, phage-liposome nanoconjugates and bio-responsive hydrogels, demonstrating significantly enhanced bactericidal efficiency compared to unmodified free phages. These bioengineered vectors attenuate bacterial virulence and resensitize pathogens to antibiotics by delivering CRISPR-Cas systems or base editors to disrupt critical virulence factors such as pili, capsule synthesis machineries and quorum sensing systems, or by inactivating antibiotic resistance determinants including beta-lactamase genes. As an intelligent nanomedicine delivery platform, engineered bacteriophage enable precise pathogen elimination an through photocatalytic reactive oxygen species generation, immunomodulatory interventions, or controlled release of antibacterial drugs. Furthermore, oral administration of engineered bacteriophage facilitates microbiota modulation, which selectively eliminate intestinal pathogens while preserve beneficial commensal microbiota, thereby restoring microbial community balance and preventing complications associated with dysbiosis. Finally, we critically analyze persistent challenges including host strain matching complexity, evolution of bacterial resistance mechanisms, pharmacokinetic optimization requirements, optimal administration route selection, large-scale production quality control standards and clinical dosing determination protocols. Through multidisciplinary integration of synthetic biology, infectious disease medicine and immunology, future translational medicine studies of bacteriophage should establish comprehensive technical platforms encompassing rapid phage screening, intelligent rational design, rigorous in vivo evaluation and standardized clinical validation processes, ultimately advancing engineered bacteriophage from laboratory innovations to clinically approved therapeutics for effectively combating MDR bacterial infections.

Objective: To evaluate the feasibility of dynamic contrast-enhanced MRI (DCE-MRI) in differentiating tumor deposits (TDs) from metastatic lymph nodes (MLNs) in rectal cancer. Materials and Methods: A retrospective analysis was conducted on 70 patients with rectal cancer, including 168 lesions (70 TDs and 98 MLNs confirmed by histopathology), who underwent pretreatment MRI and subsequent surgery between March 2019 and December 2022. The morphological characteristics of TDs and MLNs, along with quantitative parameters derived from DCE-MRI (K trans , kep, and v e) and DWI (ADCmin, ADCmax, and ADCmean), were analyzed and compared between the two groups.Multivariable binary logistic regression and receiver operating characteristic (ROC) curve analyses were performed to assess the diagnostic performance of significant individual quantitative parameters and combined parameters in distinguishing TDs from MLNs. Results: All morphological features, including size, shape, border, and signal intensity, as well as all DCE-MRI parameters showed significant differences between TDs and MLNs (all P < 0.05). However, ADC values did not demonstrate significant differences (all P > 0.05). Among the single quantitative parameters, v e had the highest diagnostic accuracy, with an area under the ROC curve (AUC) of 0.772 for distinguishing TDs from MLNs. A multivariable logistic regression model incorporating short axis, border, v e, and ADC mean improved diagnostic performance, achieving an AUC of 0.833 (P = 0.027). Conclusion The combination of morphological features, DCE-MRI parameters, and ADC values can effectively aid in the preoperative differentiation of TDs from MLNs in rectal cancer.

Objective: To evaluate the feasibility of dynamic contrast-enhanced MRI (DCE-MRI) in differentiating tumor deposits (TDs) from metastatic lymph nodes (MLNs) in rectal cancer. Materials and Methods: A retrospective analysis was conducted on 70 patients with rectal cancer, including 168 lesions (70 TDs and 98 MLNs confirmed by histopathology), who underwent pretreatment MRI and subsequent surgery between March 2019 and December 2022. The morphological characteristics of TDs and MLNs, along with quantitative parameters derived from DCE-MRI (K trans , kep, and v e) and DWI (ADCmin, ADCmax, and ADCmean), were analyzed and compared between the two groups.Multivariable binary logistic regression and receiver operating characteristic (ROC) curve analyses were performed to assess the diagnostic performance of significant individual quantitative parameters and combined parameters in distinguishing TDs from MLNs. Results: All morphological features, including size, shape, border, and signal intensity, as well as all DCE-MRI parameters showed significant differences between TDs and MLNs (all P < 0.05). However, ADC values did not demonstrate significant differences (all P > 0.05). Among the single quantitative parameters, v e had the highest diagnostic accuracy, with an area under the ROC curve (AUC) of 0.772 for distinguishing TDs from MLNs. A multivariable logistic regression model incorporating short axis, border, v e, and ADC mean improved diagnostic performance, achieving an AUC of 0.833 (P = 0.027). Conclusion The combination of morphological features, DCE-MRI parameters, and ADC values can effectively aid in the preoperative differentiation of TDs from MLNs in rectal cancer.

Nitrogen narcosis is a neurological syndrome that manifests when humans or animals encounter hyperbaric nitrogen, resulting in a range of motor, emotional, and cognitive abnormalities. The anterior cingulate cortex (ACC) is known for its significant involvement in regulating motivation, cognition, and action. However, its specific contribution to nitrogen narcosis-induced hyperlocomotion and the underlying mechanisms remain poorly understood. Here we report that exposure to hyperbaric nitrogen notably increased the locomotor activity of mice in a pressure-dependent manner. Concurrently, this exposure induced heightened activation among neurons in both the ACC and dorsal medial striatum (DMS). Notably, chemogenetic inhibition of ACC neurons effectively suppressed hyperlocomotion. Conversely, chemogenetic excitation lowered the hyperbaric pressure threshold required to induce hyperlocomotion. Moreover, both chemogenetic inhibition and genetic ablation of activity-dependent neurons within the ACC reduced the hyperlocomotion. Further investigation revealed that ACC neurons project to the DMS, and chemogenetic inhibition of ACC-DMS projections resulted in a reduction in hyperlocomotion. Finally, nitrogen narcosis led to an increase in local field potentials in the theta frequency band and a decrease in the alpha frequency band in both the ACC and DMS. These results collectively suggest that excitatory neurons within the ACC, along with their projections to the DMS, play a pivotal role in regulating the hyperlocomotion induced by exposure to hyperbaric nitrogen.
Animals ; Gyrus Cinguli/drug effects* ; Male ; Mice, Inbred C57BL ; Locomotion/drug effects* ; Neurons/drug effects* ; Mice ; Nitrogen/toxicity* ; Inert Gas Narcosis/physiopathology* ; Corpus Striatum/physiopathology*

The ventral anterior (VA) nucleus of the thalamus is a major target of the basal ganglia and is closely associated with the pathogenesis of Parkinson's disease (PD). Notably, the VA receives direct innervation from the hypothalamic histaminergic system. However, its role in PD remains unknown. Here, we assessed the contribution of histamine to VA neuronal activity and PD motor deficits. Functional magnetic resonance imaging showed reduced VA activity in PD patients. Optogenetic activation of VA neurons or histaminergic afferents significantly alleviated motor deficits in 6-OHDA-induced PD rats. Furthermore, histamine excited VA neurons via H1 and H2 receptors and their coupled hyperpolarization-activated cyclic nucleotide-gated channels, inward-rectifier K⁺ channels, or Ca²⁺-activated K⁺ channels. These results demonstrate that histaminergic afferents actively compensate for Parkinsonian motor deficits by biasing VA activity. These findings suggest that targeting VA histamine receptors and downstream ion channels may be a potential therapeutic strategy for PD motor dysfunction.
Animals ; Histamine/metabolism* ; Male ; Oxidopamine/toxicity* ; Rats ; Ventral Thalamic Nuclei/physiopathology* ; Rats, Sprague-Dawley ; Disease Models, Animal ; Parkinson Disease/metabolism* ; Neurons/physiology* ; Humans ; Optogenetics