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.

ObjectiveHepatocellular carcinoma (HCC) represents 90% of all primary liver cancers. The main risk factors associated with HCC include viral hepatitis (B and/or C), alcohol abuse, and metabolic dysfunction-associated steatotic liver disease (MASLD), which progressively advance to liver fibrosis, cirrhosis, and ultimately evolve into HCC. Surgical resection represents the most effective treatment for HCC, while recent advances in immunotherapy, including immune checkpoint inhibitors and adoptive cell therapies, have provided improved treatment prospects for patients with unresectable HCC. However, the complex metabolic heterogeneity of HCC limits the therapeutic efficacy. Metabolic intermediates acyl-CoA not only provide energy and substrates for numerous biochemical reactions but also serve as donors for protein lysine acylation, a major class of post-translational modification (PTM). Therefore, a deeper understanding of the molecular mechanisms underlying protein lysine acylation and hepatocarcinogenesis is urgently needed. MethodsThe levels of protein lysine acylation and silence information regulator 5 (SIRT5) expression levels in clinical HCC samples were analyzed by Western blot. Quantitative malonylome and succinylome of HCC samples were analyzed by antibody-based affinity enrichment coupled with tandem mass spectrometry. The proliferation of HCC cells was analyzed with Cell Counting Kit-8 (CCK-8) assays, the apoptosis was quantified by Annexin V-FITC/propidium iodide (PI) staining coupled with flow cytometry, and the ability of cells to migrate was assayed by Transwell assays. The enzymatic activity of glutathione S-transferase Mu 1 (GSTM1) was quantified. Transgenic mice with hepatic overexpression of SIRT5 were constructed using CRISPR-Cas9, and primary hepatocarcinogenesis was induced by administration of diethylnitrosamine. ResultsWestern blot analysis indicated that the expression level of SIRT5 was elevated in clinical samples from HCC patients, and the levels of lysine malonylation, glutarylation, and succinylation were significantly reduced in HCC tissues. Knockout of SIRT5 in MHCC-97H and MHCC-97L hepatoma cells suppressed cell proliferation, and increased the percentage of apoptotic cells significantly. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses of the differentially malonylome and succinylome of HCC samples revealed significant enrichment in two major classes of biological processes: core energy metabolism (e.g., glycolysis/gluconeogenesis, tricarboxylic acid metabolic process, fatty acid beta oxidation) and detoxification and oxidative stress response (e.g., response to toxic substance, chemical carcinogenesis, reactive oxygen species (ROS)). SIRT5 removes malonylation from lysine residues in GSTM1 and restores its detoxification activity, which is crucial for the survival of hepatocytes under stressed conditions. More importantly, in vivo experiment indicated that hepatic-specific overexpression of SIRT5 in mice accelerated diethylnitrosamine-induced liver fibrosis and hepatocarcinogenesis, indicating the critical role of SIRT5 in HCC progression. ConclusionThis study highlights the previously unrecognized SIRT5-GSTM1 axis as a key regulator in hepatocarcinogenesis, and suggests a potential target for the treatment of patients with HCC.

Objective:A preliminary trial is conducted to explore whether perioperative(preoperative,intraoperative,and postoperative)electroacupuncture is more effective than postoperative electroacupuncture in improving gastrointestinal function for colorectal cancer patients.Methods and analysis:The study proposes a randomized,parallel-controlled,single-center trial involving 30 colorectal cancer patients aged 18-79 requiring elective surgery.Participants are randomly assigned to two groups,with equal allocation,where one group receives perioperative electroacupuncture,and the other group receives postoperative electroacupuncture.The treatment duration spans from preoperative to postoperative 72 h,with a subsequent 28-day follow-up period.The primary outcome is the time of first postoperative defecation.The secondary outcomes include the recovery time of postoperative bowel sounds,time of the first flatus,dietary recovery,postoperative gastrointestinal dysfunction frequency,quality of life scale,postoperative pain degree,time of the first ambulation,length of hospital stay,gastrointestinal hormone indicators,and adverse events.The coagulation function test,liver and renal function,and stool and blood routine serve as security biomarkers.The statistical analysis includes the t-test,rank-sum test,Chi-square test,and analysis of variance.A two-sided significance level is set at 5%.Discussion:This study aims to evaluate the feasibility and preliminary effectiveness of perioperative electroacupuncture for gastrointestinal function recovery following colorectal cancer surgery.The study's strengths include its randomized design,well-defined intervention periods,and multi-dimensional outcome assessment.Nevertheless,limitations,such as the small sample size and single-center setting,may affect external validity.The findings will guide protocol refinement and sample size estimation for future large-scale multi-center randomized controlled trials.

Objective:To observe the clinical efficacy of repetitive transcranial magnetic stimulation(rTMS)combined with mindfulness-based cognitive therapy(MBCT)in patients with alcohol withdrawal syndrome(AWS).Methods:The 120 patients with AWS who were observed in this study were all male patients admitted to our hospital from June 2021 to June 2024,the patients were divided into group A(conventional treatment,40 cases),group B(group A combined with rTMS,40 cases),and group C(group B combined with MBCT,40 cases)according to random number table method.The clinical efficacy,self-control ability[Modified Clinical Institution Alcohol Dependence Withdrawal Assessment Scale(CIWA-Ar)score,Visual Analog Scale of Psychological Craving for Alcohol(VAS)score and Pennsylvania Alcohol Craving Scale(PACS)score],anxiety and depression degree assessment[Hamilton Depression Scale(HAMD)score,Hamilton Anxiety Scale(HAMA)score]and quality of life[36 Short Form Health Survey(SF-36)Score],relapse rate and readmission rate were compared among the three groups.Results:The total effective rate of group A,group B and group C increased successively(P＜0.05).The CIWA-Ar,PACS and VAS scores in group B and group C after treatment were lower than those in group A,and group C was lower than that in group B(P＜0.05).The HAMD and HAMA scores of group B and group C after treatment were lower than those in group A,and group C was lower than that in group B(P＜0.05).The SF-36 score of group B and group C after treatment was higher than those in group A,and group C was higher than that in group B(P＜0.05).Relapse rate and readmission rate in groups B and C were lower than those in group A,and group C was lower than that in group B(P＜0.05).Conclusion:The application of rTMS combined with MBCT in patients with AWS can improve clinical efficacy and quality of life,alleviate anxiety and depression,improve patients' self-control ability,reduce relapse rate and readmission rate,with definite effects.

Objective:To improve the teaching effect and promote the cultivation of medical talents by innovation of teaching design and learning methods for the "medical ethics" course.Methods:The "medical ethics" course in fall semester of 2021 was reformed, and 779 undergraduate students participated in the course. The course reform included the following elements: "clinical application" was set as the teaching goal and senior clinicians served as the main faculty; a "student-centered" learning format was developed based on social cognitive theory and the case-based learning (CBL) + clinical-scenario-deduction teaching method; formative assessment was conducted for individuals and groups, and essays, reviews, or reports of thoughts and experiences were used as the primary method of summative assessment. Questionnaires were used to evaluate students' satisfaction with the course reform. SPSS 24.0 software was used for statistical analysis. The results of the satisfaction survey were reported as percentage (number of specific cases/total cases).Results:A total of 770 valid questionnaires were collected. The results showed that 98.83% (761/770) students were satisfied with the course, the average score of teachers was 9.5 (0-10) points, the overall satisfaction of the teaching arrangement was 80.52% (620/770), and 96.10% (740/770) students reported that the course had a positive impact on their future career planning and facilitated multiple improvements in learning outcomes.Conclusions:Through the application of appropriate education and teaching methods in the reform of "medical ethics" course, the comprehensive quality and humanistic quality of students have been significantly improved, the teaching purpose has been achieved, and high levels of student satisfaction and teaching effect have been obtained.

The course " Melody of Genes", jointly constructed by teachers from the medical and humanities and art disciplines, aims to help medical students to acquire a deep understanding of the inseparable relationship between medicine and humanities and art and improve their philosophical literacy and humanistic care spirit. Since its implementation in 2022, a total of 116 students have been enrolled over three sessions. For this course, a student-centered design is employed, an online platform is established, and multiple teaching methods (instructional videos, classroom discussions, instructor summaries, and post-class expansion with online resources) are integrated. The course assessment primarily consists of classroom discussions and students' final reports on self-selected topics, supplemented by student feedback summaries and questionnaires. According to a total of 109 valid questionnaires, through the study of this course, 101 (92.66%) students thought that it was helpful to improve humanistic and philosophical literacy, 91 (83.49%) students thought that their self-learning and critical thinking abilities were improved, and 92 (84.40%) students believed that the course enhanced their empathy for patients and professional identity as doctors. This study provides a reference for the construction and practice of interdisciplinary medical-humanities curricula under the context of new medical education.

The Lambl's excrescences(LEs)are elongated valvular fronds with excessive mobility,commonly found at the closure lines of heart valves.These filiform projections are thin and long,exhibiting undulating independent motion.LEs predominantly occur on the ventricular side of the aortic valve and on the atrial side of the mitral valve,although there have been documented cases involving tricuspid,pulmonic,and prosthetic valves.To detect LEs,transesophageal echocardiography is often recommended.The high stress experienced by left-sided valves during closure can lead to tears or lesions in the endocardium that initiate LEs pathogenesis.In this particular case study,we present a patient who presented with acute non-ST-segment elevation myocardial infarction alongside LEs located on the non-coronary cusp of the aortic valve.This specific anatomical location posed significant challenges during percutaneous coronary intervention(PCI),as it required direct contact between the guiding catheter and the aortic valve,thereby increasing the risk of dislodging the LEs.Fortunately,our team successfully performed PCI without encountering any notable complications or embolic events.

Objective:A preliminary trial is conducted to explore whether perioperative(preoperative,intraoperative,and postoperative)electroacupuncture is more effective than postoperative electroacupuncture in improving gastrointestinal function for colorectal cancer patients.Methods and analysis:The study proposes a randomized,parallel-controlled,single-center trial involving 30 colorectal cancer patients aged 18-79 requiring elective surgery.Participants are randomly assigned to two groups,with equal allocation,where one group receives perioperative electroacupuncture,and the other group receives postoperative electroacupuncture.The treatment duration spans from preoperative to postoperative 72 h,with a subsequent 28-day follow-up period.The primary outcome is the time of first postoperative defecation.The secondary outcomes include the recovery time of postoperative bowel sounds,time of the first flatus,dietary recovery,postoperative gastrointestinal dysfunction frequency,quality of life scale,postoperative pain degree,time of the first ambulation,length of hospital stay,gastrointestinal hormone indicators,and adverse events.The coagulation function test,liver and renal function,and stool and blood routine serve as security biomarkers.The statistical analysis includes the t-test,rank-sum test,Chi-square test,and analysis of variance.A two-sided significance level is set at 5%.Discussion:This study aims to evaluate the feasibility and preliminary effectiveness of perioperative electroacupuncture for gastrointestinal function recovery following colorectal cancer surgery.The study's strengths include its randomized design,well-defined intervention periods,and multi-dimensional outcome assessment.Nevertheless,limitations,such as the small sample size and single-center setting,may affect external validity.The findings will guide protocol refinement and sample size estimation for future large-scale multi-center randomized controlled trials.

Aim To explore the mechanism of baicalin combined with heat stimulation in treating acute lym-phoblastic leukemia(ALL)based on network pharma-cology and in vitro experiments.Methods The CCK-8 assay was used to screen the suitable conditions for heat stimulation to interfere ALL cell lines Jurkat,CCRF-CEM,Hut-78 and a normal lymphocyte HMy2.CIR,and the effects of baicalin combined with heat stimulation on the proliferation of three ALL cell lines and a normal lymphocyte were tested.The key targets of baicalin combined with fever stimulation for the treatment of ALL were obtained based on network phar-macological analysis,and the potential mechanisms were predicted by gene ontology(GO)annotation and kyoto encyclopedia of genes and genomes(KEGG)en-richment.The expression levels of TNF-α,AKT1,TYMS and CASP3 mRNA in ALL cell lines Jurkat and CCRF-CEM were examined by RT-qPCR with baicalin alone and baicalin combined with heat stimulation.Results The optimal conditions for heat stimulation to intervene ALL cells were 41 ℃ for 24 h,and heat stimulation combined with baicalin synergistically inhibited the growth of ALL cell lines and effectively reduced the cy-totoxicity of baicalin.Based on the network pharmaco-logical analysis,55 intersecting targets of baicalin with ALL diseases and 77 intersecting targets of baicalin with fever were obtained.The results of GO annotation and KEGG enrichment suggested that baicalin com-bined with fever stimulation to intervene ALL might be associated with influencing intracellular reactive oxygen species metabolism,DNA transcription and apoptotic processes involved in cysteine enzymes.Apoptosis,TNF and IL-17 signaling pathways were the key pathways for baicalin combined with heat stimulation in treating ALL.Under heat stimulation at 41 ℃ using SDHA gene as housekeeping gene,in vitro experiments showed that baicalin significantly up-regulated the expression of TNF-α and CASP3,and down-regulated the expression of TYMS in ALL cells.Conclusions Based on net-work pharmacologic analyses and in vitro experiments,baicalin combined with heat stimulation can regulate TNF-α and CASP3 gene levels in ALL cells and de-stroy cellular structure to promote cell apoptosis,thus synergistically treating ALL.