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.

Plant-derived extracellular vesicles (PDEVs) are nanoscale extracellular vesicles secreted by plant cells, characterized by a lipid bilayer structure. These vesicles carry a variety of bioactive molecules, including proteins, nucleic acids, and lipids, and play essential roles in intercellular communication and physiological regulation in plants. Compared to animal-derived extracellular vesicles, PDEVs offer several advantages, such as a broad range of sources, high biocompatibility, low immunogenicity, and low production costs. Furthermore, PDEVs have demonstrated remarkable potential as natural nanocarriers for drug delivery, due to their ability to efficiently traverse biological barriers, such as the blood-brain barrier, making them promising candidates for drug delivery systems. This review systematically elaborates on the complex composition of PDEVs, which consists of lipids, proteins, and nucleic acids, the typical structural characteristics of their lipid bilayers ranging from 30 to 150 nm, and their versatile loading capabilities as drug carriers, efficiently encapsulating various types of therapeutic agents such as hydrophilic small molecules, hydrophobic drugs, nucleic acids, and proteins. We systematically summarize the recent advancements in strategies for enhancing the loading efficiency of PDEVs, which include methods such as co-incubation, ultrasound-assisted loading, electroporation, freeze-thaw cycles, and microfluidic technology. These techniques are evaluated based on their underlying principles, suitable drug types, and their respective advantages. In addition to loading strategies, we focus on the engineered approaches to achieve targeted delivery using PDEVs, such as genetic engineering modifications, chemical ligand conjugation, membrane fusion technology, and polyethylene glycol (PEG) modification. We discuss the mechanisms of these strategies in enhancing targeting efficiency, prolonging in vivo circulation time, and improving therapeutic efficacy. Further, this review highlights the application of PDEVs in various disease models, including tumor, skin inflammation, metabolic disorders, and neurodegenerative diseases, showcasing their therapeutic potential as multifunctional delivery platforms. The ability of PDEVs to encapsulate diverse therapeutic agents and target specific tissues or cells opens up new avenues for the treatment of complex diseases, offering advantages over conventional drug delivery systems. However, despite the promising applications of PDEVs, several challenges remain in their development and clinical translation. These challenges include variability in source materials, standardization of preparation processes, quality control, scalability of production, and the need for clinical validation. To overcome these obstacles, the integration of advanced technologies such as artificial intelligence-assisted design and multi-omics analysis is proposed as a way to facilitate the precise development of PDEVs. These emerging technologies hold the potential to further enhance the precision and effectiveness of plant-based drug delivery systems, ultimately advancing the field of precision medicine. In conclusion, the use of PDEVs as a platform for drug delivery represents a promising area of research with the potential to revolutionize therapeutic strategies. Their ability to encapsulate and deliver a wide variety of bioactive molecules, along with their inherent advantages in biocompatibility and versatility, makes them a valuable tool in the development of more efficient and targeted therapeutic interventions. Continued research and innovation in this field will pave the way for the clinical implementation of PDEVs in the treatment of various diseases, offering new hope for more effective and sustainable therapeutic options.

Objective To investigate the perioperative management of antithrombotic drugs in patients undergoing non-cardiac surgery.Methods Patients on long-term antithrombotic drugs who underwent non-cardiac surgery in our hospital were included.Through interviews with patients and physicians,perioperative antithrombotic medication regimens were reviewed and compared with the"Multidisciplinary Expert Consensus on Perioperative Management of Antithrombotic Therapy"to evaluate compliance with consensus and analyze influencing factors.Results A total of 372 patients were included in the analysis.Among them,355 patients were on long-term antiplatelet therapy alone,and 17 were on long-term oral anticoagulantion.364(97.8％)discontinued antithrombotic medications prior to surgery.109 patients(29.3％)received low molecular weight heparin(LMWH)bridging therapy.Among the 355 patients on antiplatelet therapy,108(30.4％)had discontinuation durations consistent with the consensus recommendations,while 186(52.4％)discontinued medications for longer periods.Postoperatively,the average hospital stay for antiplatelet therapy patients was 6.64 days,with only 37(10.4％)resuming therapy during hospitalization.The average hospital stay for patients on anticoagulants was 9.94 days,with only 2(11.8％)resuming therapy during hospitalization.Regarding perioperative risk assessment,only 40(10.8％)of patients underwent additional internal medical evaluation for thromboembolic risk after medication discontinuation,with the remainder assessed soly by surgeons.Coronary heart disease was an independent risk factor associated with internal medical evaluation(OR 2.851,95％CI 1.160-7.011,P=0.022).For bleeding risk assessment,surgeons evaluations aligned with the consensus in 68.0％of cases,but surgeons tended to underestimate risk compared to the consensus.Conclusions In this single-center study,perioperative antithrombotic management showed low compliance with expert consensus,characterized by prolonged preoperative medication discontinuation,high rates of LMWH bridging,and low postoperative in-hospital resumption of therapy.A robust multidisciplinary collaboration system should be established to enhance comprehensive patient assessment.

Aim To investigate the therapeutic effects of a newly developed Tadalafil tablets on pathological myocardial hypertrophy induced by abdominal aortic constriction(AAC)in rats,as well as its influence on the activation of the NF-κB signaling pathway in myo-cardial cells.Methods SD rats were randomly divid-ed into 4 groups:the sham operation group(Sham),the model group(AAC),the tadalafil new tablet treat-ment group(N-Tad,5 mg·kg-1),and the positive control drug treatment group(Cialis,10 mg·kg-1g).The AAC model group and treatment group rats under-went blunt dissection and constrictive ligation of the abdominal aorta at the left renal artery branch point during surgery,while the Sham group rats only had their arteries separated without any constrictive liga-tion.Rats in the treatment groups received either N-Tad or Cialis via gavage three days after modeling,while rats in the sham group and the model group re-ceived physiological saline daily for 8 weeks.Small an-imal ultra-high-resolution echocardiography and hemo-dynamic assessment were applied to evaluate left ven-tricular function in each group of rats,and the calcula-tion of the left ventricular mass index was conducted.By employing Western blot and RT-PCR.we assessed the impact of this treatment on the expression of the hy-pertrophy factor atrial natriuretic peptide(ANP),phosphorylated NF-κB p65 protein(p-NF-κB p65),and phosphorylated IκB-α in the left heart tissue of rats and in H9c2 cardiomyocytes.Results Compared to the Sham group,the AAC rats exhibited a significant decrease in left heart function,an increase in left ven-tricular mass index,and a notable increase in ANP and p-p65 expression in the left heart tissue(P＜0.05).Both N-Tad and Cialis treatments could significantly enhance left ventricular function,decrease left ventric-ular mass index,and inhibit the expression of ANP and phosphorylated NF-κB p65 in rats with myocardial hy-pertrophy(P＜0.05).Notably,the therapeutic effect of low-dose N-Tad was comparable to that of high-dose Cialis.At the cellular level,Tadalafil significantly in-hibited the activation of the NF-κB signaling pathway and reduced the expression of associated proteins in H9c2 cardiomyocytes.Conclusions N-Tad can sig-nificantly inhibit p65 and IκB-α phosphorylation,and the activation of the NF-κB signaling pathway,reduce ANP expression,and improve pathological myocardial hypertrophy,as well as mitigate left heart function damage caused by abdominal aortic constriction.

The structure and function of mitochondria and endoplasmic reticulum(ER)are important for maintaining cellular homeostasis.It has been found that the interaction between mitochondria and ER is involved in the occurrence and development of a variety of diseases.The mitochondria-associated ER membrane(MAM)is a membrane contact site between the ER and mitochondria,and is an important communication center between organelles in eukaryotic cell.Calcium channels on the ER side and the mitochondrial side are crucial in the calcium transport process in MAM.The interaction between ER and mitochondria controls mitochondrial biological function and cell survival through calcium transport regula-tion,and are involved in the occurrence and development of various pathologic process.On the one hand,MAM regulates calcium transport,which is involved in the modulation of various cellular survival and death processes.It plays a profound regulatory role in the damage of tumor cells,neuronal cells,car-diomyocytes,endothelial cells and nucleus pulposus cells through different key molecules within MAM.On the other hand,the regulation of MAM in calcium transport is crucial in the development of mitochon-drial dysfunction in Hepa 1-6 cells,the synthesis and secretion of pancreatic β-cells and amyotrophic lat-eral sclerosis.In addition,MAM also affects cellular transcription processes by regulating calcium trans-port,thereby exerting significant regulatory effects on angiogenesis and breast cancer.This paper reviews the structural features and pathophysiologic role of calcium transport regulation of MAM,and expects to provide new horizons for prevention and treatment of related diseases targeting MAM.

OBJECTIVE: To evaluate the immediate effect of Kuanxiong Aerosol (KXA) on perioperative coronary microcirculation in patients with unstable angina (UA) suffering from elective percutaneous coronary intervention (PCI). METHODS: From February 2021 to July 2023, UA inpatients who underwent PCI alone in the left anterior descending (LAD) branch were included. Random numbers were generated to divide patients into the trial group and the control group at a ratio of 1:1. The index of coronary microcirculation resistance (IMR) was measured before PCI, and the trial group was given two sprays of KXA, while the control group was not given. IMR was measured again after PCI, cardiac troponin I (cTnI) and creatine kinase isoenzyme-MB (CK-MB) were detected before and 24 h after surgery, and major cardiovascular adverse events (MACEs) were recorded for 30 days. The data statistics and analysis personnel were blinded. RESULTS: Totally 859 patients were screened, and 62 of them were involved into this study. Finally, 1 patient in the trial group failed to complete the post-PCI IMR and was excluded, 30 patients were included for data analysis, while 31 patients in the control group were enrolled in data analysis. There was no significant difference in baseline data (age, gender, risk factors, previous history, biochemical index, and drug therapy, etc.) between the two groups. In addition, differences in IMR, cTnI and CK-MB were not statistically significant between the two groups before surgery. After PCI, the IMR level of the trial group was significantly lower than that of the control group (19.56 ± 14.37 vs. 27.15 ± 15.03, P=0.048). Besides, the incidence of perioperative myocardial injury (PMI) was lower in the trial group, but the difference was not statistically significant (6.67% vs. 16.13%, P=0.425). No MACEs were reported in either group. CONCLUSIONS KXA has the potential of improving coronary microvascular dysfunction. This study provides reference for the application of KXA in UA patients undergoing elective PCI. (Registration No. ChiCTR2300069831).
Humans ; Percutaneous Coronary Intervention ; Male ; Microcirculation/drug effects* ; Female ; Angina, Unstable/physiopathology* ; Pilot Projects ; Middle Aged ; Aged ; Drugs, Chinese Herbal/pharmacology* ; Aerosols ; Troponin I/blood* ; Coronary Circulation/drug effects* ; Elective Surgical Procedures

OBJECTIVE: To evaluate the dynamic changes of glucocorticoid (GC) dose and the feasibility of GC discontinuation in rheumatoid arthritis (RA) patients under the background of Chinese medicine (CM). METHODS: This multicenter retrospective cohort study included 1,196 RA patients enrolled in the China Rheumatoid Arthritis Registry of Patients with Chinese Medicine (CERTAIN) from September 1, 2019 to December 4, 2023, who initiated GC therapy. Participants were divided into the Western medicine (WM) and integrative medicine (IM, combination of CM and WM) groups based on medication regimen. Follow-up was performed at least every 3 months to assess dynamic changes in GC dose. Changes in GC dose were analyzed by generalized estimator equation, the probability of GC discontinuation was assessed using Kaplan-Meier curve, and predictors of GC discontinuation were analyzed by Cox regression. Patients with <12 months of follow-up were excluded for the sensitivity analysis. RESULTS: Among 1,196 patients (85.4% female; median age 56.4 years), 880 (73.6%) received IM. Over a median 12-month follow-up, 34.3% (410 cases) discontinued GC, with significantly higher rates in the IM group (40.8% vs. 16.1% in WM; P<0.05). GC dose declined progressively, with IM patients demonstrating faster reductions (median 3.75 mg vs. 5.00 mg in WM at 12 months; P<0.05). Multivariate Cox analysis identified age <60 years [P<0.001, hazard ratios (HR)=2.142, 95% confidence interval (CI): 1.523-3.012], IM therapy (P=0.001, HR=2.175, 95% CI: 1.369-3.456), baseline GC dose ⩽7.5 mg (P=0.003, HR=1.637, 95% CI: 1.177-2.275), and absence of non-steroidal anti-inflammatory drugs use (P=0.001, HR=2.546, 95% CI: 1.432-4.527) as significant predictors of GC discontinuation. Sensitivity analysis (545 cases) confirmed these findings. CONCLUSIONS RA patients receiving CM face difficulties in following guideline-recommended GC discontinuation protocols. IM can promote GC discontinuation and is a promising strategy to reduce GC dependency in RA management. (Trial registration: ClinicalTrials.gov, No. NCT05219214).
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Arthritis, Rheumatoid/drug therapy* ; Glucocorticoids/therapeutic use* ; Medicine, Chinese Traditional ; Retrospective Studies

Mutations in the cyclin-dependent kinase-like 5 gene (CDKL5) cause a severe neurodevelopmental disorder, yet the impact of truncating mutations remains unclear. Here, we introduce the Cdkl5^492stop mouse model, mimicking C-terminal truncating mutations in patients. 492stop/Y mice exhibit altered dendritic spine morphology and spontaneous seizure-like behaviors, alongside other behavioral deficits. After creating cell lines with various Cdkl5 truncating mutations, we found that these mutations are regulated by the nonsense-mediated RNA decay pathway. Most truncating mutations result in CDKL5 protein loss, leading to multiple disease phenotypes, and offering new insights into the pathogenesis of CDKL5 disorder.
Animals ; Disease Models, Animal ; Mice ; Protein Serine-Threonine Kinases/deficiency* ; Mutation/genetics* ; Epileptic Syndromes/genetics* ; Humans ; Dendritic Spines/pathology* ; Spasms, Infantile/genetics* ; Male ; Seizures/genetics* ; Mice, Inbred C57BL