Objective: To explore the setting of gray zone of Treponema pallidum (TP) testing by retrospective analysis of blood donors with single reagent reactive anti-TP results, so as to improve blood utilization and supply safety. Methods: Blood samples were collected from 112 blood donors previously deferred due to single reagent reactive TP antibody results between January 2020 and December 2023, and subjected to dual ELISA reagents and TPPA test. The gray zone panel analysis was performed on the two ELISA reagents currently used in our department. The detection rate at each concentration of the gray zone panle was counted, and the corresponding concentrations for C , C , and C and gray zone cut-off were calculated. Results: Among the 50 samples deferred by reagent 1, 19 were confirmed reactive and 31 non-reactive in supplementary testing. Among the 62 samples deferred by reagent 2, 12 were confirmed reactive and 50 non-reactive in supplementary testing. For reagent 1, the detection rate of was 56% for S/CO≥1 and 20% for 0.5≤S/CO<1, retrospectively. For reagent 2, the detection rate was 27% for S/CO≥1 and 12.5% for 0.5≤S/CO<1, retrospectively. The detection rate for S/CO≥1 was higher than those for 0.5≤S/CO<1 for both reagents. All the 112 samples were negative in TPPA test. The C concentration of reagent 1 was 1.51 mIU/mL, and the concentration range of C ±20% was 1.21-1.81 mIU/mL. The C concentration of reagent 2 was 1.45 mIU/mL, and the concentration range of C ±20% was 1.16-1.74 mIU/mL. The C and C concentration of both reagents were within the C ±20% range, suggesting that the gray zone cutoff for both Reagent 1 and Reagent 2 should be set at S/CO=0.8 (80% of the CO value). Conclusion: All anti-TP single reagent reactive samples with S/CO value within the gray zone was tested negative by TPPA. It is necessary to consider the rationality and necessity of establishing the gray zone, so as to ensure blood safety and improve the utilization rate of blood resources.

With the rising incidence of diabetes， diabetic kidney disease （DKD） has become a significant global health burden. Although current prevention and treatment strategies can partially delay the progression of DKD， the risk of patients advancing to end-stage renal disease remains high. Since the concept of the "gut-kidney axis" was first introduced at the International Congress on Dialysis in 2011， research on the role of gut microbiota in the pathogenesis of DKD has received increasing attention. This review summarizes the current research on gut microbiota， explores the mechanisms through which it contributes to DKD development， and outlines clinical approaches for DKD prevention and treatment based on the "gut-kidney axis" theory. Evidence indicates that dietary interventions， intake of probiotics or prebiotics， use of metformin and novel antidiabetic drugs， and application of traditional Chinese medicine （TCM） compound formulas can effectively improve gut microbiota composition， influence metabolite production， and restore the intestinal mucosal barrier. These interventions can further regulate intestinal innate immunity and inflammatory responses， thereby modulating the progression of DKD. Despite challenges posed by the traditional oral administration of water-decocted TCM compound formulas and the complexity of their ingredients， increasing evidence suggests that TCM may indirectly affect the occurrence and development of DKD by modulating gut microbiota. This finding provides a new perspective on the potential mechanisms of TCM in DKD treatment and may offer novel strategies for DKD prevention and therapy.

Atherosclerosis （AS） is the main pathological basis of cardiovascular diseases， and its occurrence and development are closely related to epigenetic regulation. Histone modification， as the core mechanism of epigenetic regulation， plays a crucial role in the occurrence and development of AS by dynamically regulating chromatin structure and gene expression. In recent years， traditional Chinese medicine （TCM） and its active components have shown unique advantages and potential in regulating histone modification for the prevention and treatment of AS. This article systematically reviews the mechanisms of histone acetylation， methylation， lactylation， etc. in the pathological process of AS and summarizes the latest research progress in the intervention of AS by the active components and compound prescriptions of TCM through regulating histone modification. Studies have indicated that TCM and its compound prescriptions regulate the activities of histone modification enzymes via multiple targets， showing unique advantages in influencing lipid metabolism and inflammatory response， as well as stabilizing vulnerable plaques and endothelial function. The active components of TCM directly target the activities or expression of histone modification enzymes such as histone acetyltransferases （HATs）， histone deacetylases （HDACs）， histone lysine methyltransferases （KMTs）， and histone lysine demethylases （KDMs）， thereby causing changes in histone modification and ultimately affecting gene expression and pathological phenotype in AS. However， current research still has problems such as insufficient in-depth basic research， unclear intervention effects and mechanisms of AS dynamics， relatively isolated studies on various factors of epigenetic modification， and unclear establishment of quality control standards for TCM compound prescriptions based on epigenetic regulation. In the future， in-depth research is still needed， and the research results should be translated into clinical applications. This article systematically clarifies the key role and mechanism of TCM in regulating the pathological process of AS through epigenetic intervention， providing new ideas for the modernization of TCM and precise prevention and treatment of AS.

Atherosclerosis （AS） is the main pathological basis of cardiovascular diseases， and its occurrence and development are closely related to epigenetic regulation. Histone modification， as the core mechanism of epigenetic regulation， plays a crucial role in the occurrence and development of AS by dynamically regulating chromatin structure and gene expression. In recent years， traditional Chinese medicine （TCM） and its active components have shown unique advantages and potential in regulating histone modification for the prevention and treatment of AS. This article systematically reviews the mechanisms of histone acetylation， methylation， lactylation， etc. in the pathological process of AS and summarizes the latest research progress in the intervention of AS by the active components and compound prescriptions of TCM through regulating histone modification. Studies have indicated that TCM and its compound prescriptions regulate the activities of histone modification enzymes via multiple targets， showing unique advantages in influencing lipid metabolism and inflammatory response， as well as stabilizing vulnerable plaques and endothelial function. The active components of TCM directly target the activities or expression of histone modification enzymes such as histone acetyltransferases （HATs）， histone deacetylases （HDACs）， histone lysine methyltransferases （KMTs）， and histone lysine demethylases （KDMs）， thereby causing changes in histone modification and ultimately affecting gene expression and pathological phenotype in AS. However， current research still has problems such as insufficient in-depth basic research， unclear intervention effects and mechanisms of AS dynamics， relatively isolated studies on various factors of epigenetic modification， and unclear establishment of quality control standards for TCM compound prescriptions based on epigenetic regulation. In the future， in-depth research is still needed， and the research results should be translated into clinical applications. This article systematically clarifies the key role and mechanism of TCM in regulating the pathological process of AS through epigenetic intervention， providing new ideas for the modernization of TCM and precise prevention and treatment of AS.

From the theoretical perspective of "body fluids and blood stasis mixing"， environmental microplastics （MPs） are conceptualized as a "novel turbid-toxin". This study aims to elucidate the complete pathogenic pathway through which MPs act as a key driving force （the "crucible" of pathogenesis） in the initiation and progression of atherosclerosis （AS）. By tracing the classical theories in the Chapter The Occurrence of All Diseases of Miraculous Pivot （Ling Shu）， this paper clarifies the core connotations of "body fluids"-it not only refers to endogenous pathological fluids and lipid turbidity but also provides a theoretical basis for incorporating "exogenous turbid fluids"， thereby laying a logical foundation for conceptualizing MPs as a "novel turbid-toxin". Meanwhile， the implications of "blood" （encompassing both blood quality abnormalities and blood stasis） and the dynamic process of "mixing" are elucidated. Drawing upon modern toxicological evidence， this paper demonstrates the high homology between MPs and "exogenous turbid-fluids" from three aspects： Morphology， toxicity， and invasion routes. The micro/nano-scale particle morphology of MPs enables mobility within the bloodstream. The multiple exposure pathways of MPs correspond to the traditional Chinese medicine understanding of pathogens invading through the mouth， nose， and skin. The characteristics of accumulating in vivo while inducing oxidative stress and inflammatory responses of MPs fully embody the pathogenic features-adhesion， binding， and vessel damage-of "turbid-toxin". On this basis， the dynamic pathogenesis of MP-induced AS is systematically interpreted. Initially， MPs with the "turbid-toxin" nature impair nutrient-defense harmony and cause endothelial dysfunction. Subsequently， as the core of "mixing"， they interact with blood lipids and immune cells， generating heat and phlegm to form a major pathological hub of chronic inflammation. Ultimately， this process drives the coalescence of phlegm， stasis， and turbid-toxin into tangible plaques， evolving from stable lesions to vulnerable masses and accumulations. By integrating classical pathogenic model with contemporary environmental medicine， this study establishes an analytical framework that bridges macro-theory and micro-mechanisms for understanding the cardiovascular risks of MPs through an integrative Chinese-Western medicine lens.

OBJECTIVE: Benzylisoquinoline alkaloids (BIAs) have pharmacological functions and clinical use. BIAs are mainly distributed in plant species across the order Ranunculales and the genus Phellodendron from Sapindales. The BIA biosynthesis has been intensively investigated in Ranunculales species. However, the accumulation mechanism of BIAs in Phellodendron is largely unknown. The aim of this study is to unravel the biosynthetic pathways of BIAs in Phellodendron amurens. METHODS: The transcriptome and metabolome data from 18 different tissues of P. amurense were meticulously sequenced and subsequently subjected to a thorough analysis. Weighted gene co-expression network analysis (WGCNA), a powerful systems biology approach that facilitates the construction and subsequent analysis of co-expression networks, was utilized to identify candidate genes involved in BIAs biosynthesis. Following this, recombinant plasmids containing candidate genes were expressed in Escherichia coli, a widely used prokaryotic expression system. The purpose of this genetic engineering endeavor was to express the candidate genes within the bacteria, thereby enabling the assessment of the resultant enzyme activity. RESULTS: The synonymous substitutions per synonymous site for paralogs indicated that at least one whole genome duplication event has occurred. The potential BIA biosynthetic pathway of P. amurense was proposed, and two PR10/Bet v1 members, 14 CYP450s, and 33 methyltransferases were selected as related to BIA biosynthesis. One PR10/Bet v1 was identified as norcoclaurine synthase, which could catalyze dopamine and 4-hydroxyphenylacetaldehyde into (S)-norcoclaurine. CONCLUSION Our studies provide important insights into the biosynthesis and evolution of BIAs in non-Ranunculales species.

AIM:To investigate the effect of dyes,Remazol BrOrange yellow(RBY)and erythrosine(ERY),on the outcomes of immunoblotting analysis when used for staining the concentrate gel in sodium dodecyl sulfate-polyacryl-amide gel electrophoresis(SDS-PAGE).METHODS:Polyacrylamide gels were divided into five groups:the control group(prepared according to the conventional kit protocol),the RBY-stained group with a final concentration of 0.08 g/L,the RBY-stained group with a final concentration of 0.16 g/L,the ERY-stained group with a final concentration of 0.08 g/L,and the ERY-stained group with a final concentration of 0.8 g/L.Gels were prepared and subjected to electro-phoresis,followed by coomassie brilliant blue staining to visualize protein bands.Subsequently,proteins were transferred to PVDF membranes,which were then blocked,incubated with primary and secondary antibodies,washed,and finally ex-posed for imaging to observe the target protein vinculin bands.RESULTS:Compared with the unstained concentrate gel,the loading wells of the RBY or ERY pre-stained concentrate gel were more clearly visible.Analysis of the gels stained with coomassie brilliant blue after electrophoresis and marker visualization showed no significant different in protein elec-trophoretic mobility between prestained and unstained gels.Comparative analysis of the immunoblotting also indicated that the detection of protein samples transferred to PVDF membranes was unaffected.CONCLUSION:Prestaining concen-trate gels with RBY or ERY can enhance the efficiency of gel-based electrophoresis and immunoblotting analysis.

In traditional Chinese medicine,static blood refers to the pathological product formed by poor blood circulation or blood stagnation in the body.Static blood is viscous and prone to clotting,which can cause various diseases.Extracellular matrix(ECM)is a complex extracellular network structure secreted by cells.Under pathological conditions,abnormal changes occur in the components and structure of the ECM,leading to tissue hardening and fibrosis,which in turn hinders the normal functions of cells and contributes to the occurrence and development of various diseases such as atherosclerosis.Some abnormal states of ECM are consistent with the characteristics of static blood,and ECM may serve as a modern material base for static blood.The association between static blood and ECM was explored in terms of their unity in causes,composition,physical properties,distribution,and biological processes.The physiological characteristics and pathological evolution processes in the body of both are similar.Abnormal ECM deposition is the early basis for the formation and retention of atherosclerosis.Dynamic remodeling of the ECM is a key factor in the injury and rupture of atherosclerotic plaques.Abnormal ECM cross-linking aggravates the inflammatory microenvironment of atherosclerotic vessels.As abnormal pathological products and pathogenic factors,the retention of static blood and ECM deposition and remodeling are closely associated with the occurrence and development of atherosclerotic cardiovascular diseases,such as myocardial ischemia,acute myocardial infarction,and ischemic stroke.This article explores the relationship between static blood in traditional Chinese medicine and ECM,aiming to provide a new direction for the theoretical research of blood stasis,novel clinical treatment ideas,and potential intervention targets for the prevention and treatment of atherosclerotic cardiovascular diseases.

OBJECTIVE To study regularity of different components whose skin permeability is enhanced by essential oils(EOs)from traditional Chinese medicine(TCM)with hot property.METHODS Five EOs from TCM with hot property,namely Galangal oil,Dried Ginger oil,Cinnamon oil,Pepper oil,and Evodia oil were prepared and used in further studies.The in vitro skin cytotoxici-ty of these EOs and chemical penetration enhancer(PE)Azone were compared.HPLC method was established to determine 9 TCM components that are usually used in permeation studies.And the in vitro permeation experiments were carried out using the modified Franz diffusion cell method to evaluate the penetration enhancement effects of five EOs.RESULTS The cytotoxicity test revealed the IC50 value of EOs was 3.63-8.15 times of that of Azone,the classical chemical PE.HPLC method showed perfect specificity and 9 TCM components performed well in linear relationship,precision,repeatability,stability,and average recovery rate.The results of multiple linear regression showed a significant association between the penetration enhancement effects of EOs from TCM with hot prop-erty and log P values of the TCM components.EOs from TCM with hot property showed satisfactory penetration enhancement effects for hydrophobic components with log P values in the range of 2.6-3.5,e.g.resveratrol,tetrahydropalmatine and quercetin.CONCLU-SION EOs from TCM with hot property should be reasonably selected as PE according to the oil-water partition coefficient of the in-gredients.