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.

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.

Aim To explore the mechanism by which the compound Chaijin Jieyu formula(CCJJY)regulates the TLR4/NLRP3 signaling pathway to inhibit central oxidative stress and improve hippocampal synaptic plasticity damage in depression.Methods SD rats were randomly divided into the control group,chronic unpredictable mild stress group,sleep deprivation group,chronic unpredictable mild stress combined with sleep deprivation group,positive drug group(venlafax-ine+melatonin),low-dose group of CCJJY,medium dose group of CCJJY,and high-dose group of CCJJY,with nine rats in each group.Except for the control group,a rat model of depression complicated with in-somnia was established using chronic unpredictable mild stress combined with sleep deprivation.Depres-sion-like and sleep behaviors in rats were evaluated through weight,food intake,water maze,and pento-barbital sodium tests.ELisa was used to detect ROS,AANAT,and HPLC-EC was used to detect 5-HT con-tent,while Western blot/RT-PCR was used to detect the expression of IL-1β,TLR4,NLRP3,PSD-95,and SYN related proteins and mRNA.HE and Golgic stai-ning were used to observe the pathological changes in the third ventricle,hippocampus,and neuronal synap-ses.Results Compared with the control group,the depression-like behaviors of the model group rats were significant.The expression of IL-1β,TLR4,and NL-RP3 in the hippocampus increased,while the expres-sion of PSD-95 and SYN decreased.Activation of NL-RP3 inflammasomes led to "sleeve like" pathological changes in the third ventricle,with hippocampal neu-rons undergoing apoptosis and significant damage to neuronal synaptic plasticity.Compared with the model group,after intervention with CCJJY,the expression of ROS,IL-1β,TLR4,and NLRP3 decreased,while the expression of AANAT,5-HT,PSD-95,and SYN in-creased.Pathological damage to the third ventricle and hippocampal neurons was repaired.Conclusion The CCJJY improves hippocampal synaptic plasticity dam-age in depression by regulating the TLR4/NLRP3 sig-naling pathway to inhibit central oxidative stress.

Acute ischemic stroke(AIS)is associated with high mortality and disability rates,presenting a substantial challenge to global public health challenge.Intravenous thrombolysis(IVT)is recognized as a cornerstone of early AIS treatment and is recommended as the standard therapeutic approach by both national and international guidelines.However,the clinical efficacy of IVT remains suboptimal due to several limitations,including a narrow therapeutic time window and the inevitable activation of the coagulation system and platelet aggregagation during thrombolysis.These factors may contribute to adverse outcomes such as early neurological deterioration(END)and vascular re-occlusion.Antiplatelet therapy(APT),which inhibits platelet aggregations,reduces microthrombus formation,and stabilizes the vascular endothelium with multifaceted mechanisms,has emerged as a promising adjunctive strategy to IVT,offering potential synergistic effects.This review summarized the latest evidence from both domestic and international studies,focusing on the mechanisms of APT,recent clinical advancements in IVT combined with APT,and the safety and efficacy of APT administration at different time windows relative to IVT.Emphasis is placed on the influence of various antiplatelet agents,dosing regimens,and initiation timing on therapeutic outcomes,alongside a comprehensive evaluation in the context of current guideline recommendations and clinical practice.Current guidelines recommend initiating APT 24 h after IVT,following imaging confirmation to exclude the risk of intracranial hemorrhage.However,the efficacy and safety of earlier APT initiation remain inconclusive.Individualized treatment strategies,such as early administration of low-dose,short-acting APT or combination therapy in specific patient subgroups,may effectively balance therapeutic benefits and risks.The adjunctive use of APT in IVT holds promise for enhancing efficacy and improving clinical outcomes,but precise stratification of safety and efficacy is essential.Future research should focus on optimizing combination IVT and APT strategies through individualized patient profiling,appropriate drug selection,and dynamic imaging monitoring to achieve precision management in AIS treatment.

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.

Family with sequence similarity 3 member A(FAM3A),a novel mitochondrial protein,plays a pivotal role in hepatic glucose and lipid metabolism by enhancing ATP synthesis and secretion and mod-ulating the ATP-P2 receptor-Akt signaling pathway.Dysregulation of FAM3A is closely associated with the pathogenesis of non-alcoholic fatty liver disease(NAFLD)and type 2 diabetes mellitus(T2DM).In this study,targeting FAM3A as a therapeutic candidate,we conducted virtual screening to identify 47 small-molecule compounds with potential binding activity.Surface plasmon resonance(SPR)analysis re-vealed three compounds exhibiting high binding affinity to FAM3 A.Further structural characterization of the FAM3A-compound complexes,combined with intermolecular interaction analysis,elucidated the binding mode of the lead compound Index 2(taxifolin)to FAM3A at atomic resolution.These findings provide critical insights into the molecular mechanisms underlying ligand-FAM3A interactions and deliver valuable chemical scaffolds for the development of therapeutics targeting NAFLD and T2DM.This work establishes a foundation for advancing drug discovery efforts focused on FAM3A-mediated metabolic disor-ders.

Objective To evaluate the efficacy and safety of transcatheter aortic valve implantation(TAVI)for the treatment of primary aortic valve regurgitation(NAVR)and to compare the difference in the choice of prosthetic valve size and the difference in complications with aortic stenosis(AS).Methods According to the definition of Valve Academic Research Consortium(VARC-3),143 patients with NAVR/AS treated with TAVI and patients with NAVR treated with surgical aortic valve replacement(SAVR)at Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,China,from March 2019 to September 2024 were selected,and clinical data on baseline,perioperative,and primary endpoint events were were retrospectively collected and compared.Results Forty-three patients with NAVR were treated with TAVI,with a device success rate of 86.0％and a surgical success rate of 95.3％.Subgroup comparisons:(1)NAVR-TAVI group than NAVR-SAVR group:patients in the TAVI group had a significantly shorter operative time than those in the SAVR group(P＜0.001);complete left bundle branch block was more likely to occur after TAVI(P=0.042),and complete right bundle branch block was more likely to occur after SAVR(P=0.044).SAVR postoperatively The incidence of congestive heart failure was higher(P=0.013),and the mortality rate was significantly higher in the SAVR group than in the TAVI group(P=0.019).(2)NAVR-TAVI group than AS-TAVI group:the differences in access selection,THV size[28(22,34)mm vs.24(22,32)mm,P=0.044]and proportion of THV overdiameter[14％(7％,20％)vs.7％(3％,11％),P＜0.001]were statistically significant.patients in AS and NAVR groups had 1 case of permanent pacing after TAVI treatment.In the AS and NAVR groups,there was 1 case of permanent pacemaker implantation after TAVI.2 patients in the AS group were converted to surgical treatment,and 6 patients died.Conclusions The use of"off-label"(transfemoral)and"on-label"(transapical)TAVI devices(both from domestic sources)is safer than SAVR for the treatment of NAVR,especially in elderly and high-risk patients.Compared with patients with AS treated with TAVI,larger diameter annulas are usually selected for NAVR,with higher rates of valve migration,but overall safety and efficacy are comparable to AS.

The pathogenesis of central precocious puberty(CPP)is due to the early initiation of the function of the hypothalamic-pituitary-gonadal axis,which is driven by the gonadotrophin-releasing hor-mone(GnRH)from the hypothalamus.The increase of GnRH secretion will promote the secretion of gon-adotropin by the pituitary gland,which leads to the development of reproductive organs and the secretion of sex hormones.The timing of puberty onset is influenced by the interaction between nutritional,envi-ronmental,and socioeconomic factors.In the study of pathogenic genes,loss-of-function mutations of MKRN3,LIN28 and DLK1 genes can lead to early onset of puberty,among which loss-of-function muta-tions of MKRN3 are one of the most common single-gene causes of familial CPP.Since 2013,when the MKRN3 mutation was first identified in five CPP families,the role of MKRN3 in adolescence has gradu-ally been discovered.MKRN3 is a member of the E3 ubiquitin ligase family and is expressed in the cen-tral nervous system of many eukaryotes.MKRN3 may act as an E3 ubiquitin ligase to inhibit GnRH activ-ity.Recently,more and more studies have explored the molecular mechanism of MKRN3 in CPP,revea-ling a variety of genes and proteins that interact with MKRN3.For example,MKRN3 can inhibit KISS1 and TAC3 activities,thereby affecting the expression of kisspeptin and neurokinin B to regulate GnRH se-cretion.MKRN3 also inhibited GNRH1 mRNA translation through PABPC1 ubiquitination.In addition,there are other targets such as MBD3,IGF2BP1 and NPTX1,all of which are involved in GnRH regula-tion downstream of MKRN3.Upstream of MKRN3,miR-30 can bind to three sites in the 3'-untranslated region of the MKRN3 gene to block MKRN3 transcription.Here we review the role of MKRN3 in the path-ogenesis of CPP and the research progress of its molecular mechanism,which will help to better under-stand the pathogenesis of CPP,provide new ideas and treatment methods for CPP-related fields in the fu-ture,and lay a theoretical foundation for further exploration of the molecular mechanism of MKRN3.

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.