ObjectiveTo explore the potential mechanism of Weicanqing Formula （微残清方， WF） combined with chemotherapy in treating acute myeloid leukemia （AML） based on malic enzyme 2 （ME2）-mediated bone marrow immune-metabolic homeostasis and . MethodsSixty-five male C57BL/6N mice were randomly divided into control group， model group， WF group， chemotherapy group， and the combination group （WF plus chemotherapy）， with 13 mice per group. Except for the control group， mice were injected with C1498 cells （appro-ximately 1×10⁶ cells per mouse） via the tail vein on day 1 to establish an AML model. Starting from day 8， mice in the chemotherapy group and the combination group were administered with cytarabine at 30 mg/（kg·d） via subcutaneous injection， once daily for 3 consecutive days； WF group and the combination group received WF at 7.54 g/（kg·d） by gavage， once daily for 21 consecutive days. On days 1， 7， 14， 21 and 28， hemoglobin （Hb） and white blood cell （WBC） levels were determined in each group. On day 28， Giemsa staining was used to observe morphological changes in bone marrow cells. Flow cytometry was employed to detect the expression levels of bone marrow T lympthocyte subsets， including CD8⁺， CD4⁺， and regulatory T lymphocytes （Treg）. The contents of glutamine （Gln）， nicotinamide adenine dinucleotide phosphate （NADP⁺）， and reduced nicotinamide adenine dinucleotide phosphate （NADPH） in bone marrow were determined using visible spectrophotometry. Adenosine triphosphate （ATP） concentration in bone marrow was measured by chemiluminescence assay. The mRNA expression of malic enzyme 2 （ME2） in bone marrow was detected by RT-qPCR， and the protein expression level of ME2 was determined by Western blotting. ResultsCompared to the control group， the model group showed decreased Hb levels on day 14， 21， and 28， and increased WBC levels on day 7， 14， 21， and 28 （P＜0.05）. On day 28， bone marrow CD8⁺ and CD8⁺/CD4⁺ levels decreased， while Treg cells， ATP， Gln， NADP⁺， NADPH， ME2 mRNA， and their corresponding protein levels increased （P＜0.05）. Compared to the model group， the chemotherapy group and the combination group both exhibited reduced Hb level on day 14， 21 and 28， as well as the increased WBC level； the combination group showed increased CD8⁺ level， decreased Treg， ATP， Gln， and NADPH content， as well as reduced ME2 mRNA expression and protein levels （P＜0.05）. Compared to the chemotherapy group， the combination group showed significantly increased Hb level on days 21 and 28， and increased WBC level on day 28 （P＜0.05）. Bone marrow smear pathology revealed that， in the model group， myeloid blast cells exhibited cytoplasmic vacuoles indicative of abnormal metabolic activity. In contrast， both the chemotherapy group and the combination group showed large numbers of metamyelocytes and band neutrophils， with only a few immature granulocytic blast cells observed. ConclusionWF may improve the prognosis of AML when used as an adjunct to chemotherapy by modulating ME2 expression， inhibiting metabolic energy competition within the bone marrow microenvironment， and reshaping the distribution of T lymphocyte subsets in the bone marrow.

The development of diabetic bladder dysfunction(DBD)is closely related to the dysregulation of the microbiotagut-bladder axis. Diabetic conditions significantly alter gut microbiota composition, causing intestinal barrier disruption, endotoxemia, and systemic inflammation. These alterations impair bladder function by activating bladder NOD-like receptor thermal protein domain associated protein 3(NLRP3)inflammasome, inducing oxidative stress, and nerve remodeling. Emerging technologies for microbiota analysis, Mendelian randomization, and targeted marker development have revealed the causal links between specific gut microbiota and DBD. Modulating gut microbiota increases short-chain fatty acids(SCFAs)production and suppresses inflammation; NLRP3 pathway targeting or direct SCFAs delivery are under clinical evaluation. The therapeutic directions for DBD include microbiota regulation therapy, application of probiotics and dietary fiber, as well as clinical translational research on the microbiota-gut-bladder axis. Engineered live bacterial therapies and nanocarrier technologies are emerging research directions. Current controversies focus on the causal role of dysbiosis, safety and long-term efficacy of individualized therapies, and technical bottlenecks. Future efforts require integrating multi-omics, organoid models, and cross-disciplinary collaboration to advance precision diagnostics and therapeutics targeting the microbiota-gut-bladder axis in DBD.

Frailty and depressive disorders exhibit a high prevalence and comorbidity rate in the elderly population.Their coexistence significantly reduces patients'quality of life,increases the risk of disability and mortality,and substantially exacerbates the socioeconomic burden.Emerging evidence indicates a significant bidirectional causal relationship between frailty and depressive disorders.The underlying comorbid mechanisms may be related to elevated levels of pro-inflammatory cytokines such as C-reactive protein,neutrophils,and white blood cells.Grey matter volume reduces in specific brain regions including the bilateral thalamus and right precentral gyrus.And abnormal hormone secretion,such as cortisol,resulting from the overactivation of the hypothalamic-pituitary-adrenal axis.Exercise interventions demonstrate positive effects in preventing and managing both frailty and depressive disorders,indicating broad application prospects.However,the underlying mechanisms require further validation.In summary,the comorbidity of frailty and depressive disorders in the elderly requires greater attention.Current evidence supports exercise intervention as an effective therapeutic strategy for improving health outcomes in this population.

The purpose of this study was to understand the epidemiological and biological character-istics of Clostridium perfringens in avian origin in Yangzhou.A total of 200 intestinal samples(90 from chickens and 110 from ducks)collected from different markets were isolated and identified by isolation and culture,morphological observation,staining microscopy,biochemical tests and other methods,antimicrobial susceptibility test by K-B paper method,and toxin type of isolated bacteria was detected by PCR.Five isolates were screened according to the characteristics of host origin,se-rotype and drug resistance,and multi-locus sequence typing analysis was carried out,and a phylo-genetic tree based on α toxin gene sequences was constructed for isolates from different sources.After morphological and molecular biological identification,a total of 61 strains of Clostridium perfringens were isolated,and the isolation rate was 30.5％,all of which were type A.The antimi-crobial susceptibility results showed that the isolate had the highest resistance rate to amikacin,followed by tetracycline and cotrimoxazole,and was more sensitive to ceftriaxone,penicillin,amoxicillin and imipenem.Multi-drug resistance is very serious,with six or more strains resistant to antibiotics emerging in every market.A total of five different ST types(STs)were found by multi-site sequence analysis,namely ST833,ST834,ST363,ST835,and ST837,and all of them were new ST types except ST363.The information from ST 363 is consistent with clinical isolate of Clostridium perfringens,suggesting that more attention needs to be paid to zoonotic diseases caused by Clostridium perfringens.The α toxin genes of the five isolates were located in four dif-ferent branches in the phylogenetic tree,indicating their genetic diversity,while the isolates from different provenances of the same source were located in the same branch,indicating that they had a closer evolutionary relationship.The Clostridium perfringens from poultry in Yangzhou are mainly type A,and with severe drug resistance and multi-drug resistance present.This research provides scientific evidence for the epidemiology and control of Clostridium perfringens.

Objective Systematic identification and analysis of the cytochrome P450(CYP450)gene family members and their functions in Chrysanthemum indicum L.to provide a basis for further studies on their roles in the flavonoid synthesis pathway.Methods Based on the genomic data of a diploid Chrysanthemum indicum L.,the members of the P450 gene family were identified and analyzed using bioinformatics methods.Moreover,the transcriptome and quasi-targeted metabolome analysis were combined to screen the flavone synthase Ⅱ(FNSII)genes from the CYP93 family,and their expression levels were tested in different tissues of C.indicum using real-time fluorescence quantitative PCR(qPCR).Results A total of 460 P450 genes were identified from the C.indicum genome,belonging to 43 families within 8 family clans.The encoded proteins ranged from 336 to 1538 amino acids in length,with a relative molecular mass between 38.01 to 175.00 kDa,and an isoelectric point between 5.61 to 9.71.Chromosome localization analysis indicated that these 460 P450 genes were unevenly distributed on nine chromosomes,with the highest number of light-responsive elements in the promoter regions.Expression pattern analysis showed that the expression levels of two FNSII genes(CindChr2G00102820.1 and CindChr8G00552890.1)were significantly higher in leaf of C.indicum than in flower.Conclusion The comprehensive analysis of the P450 gene family in C.indicum is helpful to further elucidate the function of the P450 gene family in flavonoid biosynthesis.

Objective To explore the accuracy of machine learning algorithms based on SHOX2 and RASSF1A methylation levels in predicting early-stage lung adenocarcinoma pathological types. Methods A retrospective analysis was conducted on formalin-fixed paraffin-embedded (FFPE) specimens from patients who underwent lung tumor resection surgery at Affiliated Hospital of Nantong University from January 2021 to January 2023. Based on the pathological classification of the tumors, patients were divided into three groups: a benign tumor/adenocarcinoma in situ (BT/AIS) group, a minimally invasive adenocarcinoma (MIA) group, and an invasive adenocarcinoma (IA) group. The methylation levels of SHOX2 and RASSF1A in FFPE specimens were measured using the LungMe kit through methylation-specific PCR (MS-PCR). Using the methylation levels of SHOX2 and RASSF1A as predictive variables, various machine learning algorithms (including logistic regression, XGBoost, random forest, and naive Bayes) were employed to predict different lung adenocarcinoma pathological types. Results A total of 272 patients were included. The average ages of patients in the BT/AIS, MIA, and IA groups were 57.97, 61.31, and 63.84 years, respectively. The proportions of female patients were 55.38%, 61.11%, and 61.36%, respectively. In the early-stage lung adenocarcinoma prediction model established based on SHOX2 and RASSF1A methylation levels, the random forest and XGBoost models performed well in predicting each pathological type. The C-statistics of the random forest model for the BT/AIS, MIA, and IA groups were 0.71, 0.72, and 0.78, respectively. The C-statistics of the XGBoost model for the BT/AIS, MIA, and IA groups were 0.70, 0.75, and 0.77, respectively. The naive Bayes model only showed robust performance in the IA group, with a C-statistic of 0.73, indicating some predictive ability. The logistic regression model performed the worst among all groups, showing no predictive ability for any group. Through decision curve analysis, the random forest model demonstrated higher net benefit in predicting BT/AIS and MIA pathological types, indicating its potential value in clinical application. Conclusion Machine learning algorithms based on SHOX2 and RASSF1A methylation levels have high accuracy in predicting early-stage lung adenocarcinoma pathological types.

Frailty and depressive disorders exhibit a high prevalence and comorbidity rate in the elderly population.Their coexistence significantly reduces patients'quality of life,increases the risk of disability and mortality,and substantially exacerbates the socioeconomic burden.Emerging evidence indicates a significant bidirectional causal relationship between frailty and depressive disorders.The underlying comorbid mechanisms may be related to elevated levels of pro-inflammatory cytokines such as C-reactive protein,neutrophils,and white blood cells.Grey matter volume reduces in specific brain regions including the bilateral thalamus and right precentral gyrus.And abnormal hormone secretion,such as cortisol,resulting from the overactivation of the hypothalamic-pituitary-adrenal axis.Exercise interventions demonstrate positive effects in preventing and managing both frailty and depressive disorders,indicating broad application prospects.However,the underlying mechanisms require further validation.In summary,the comorbidity of frailty and depressive disorders in the elderly requires greater attention.Current evidence supports exercise intervention as an effective therapeutic strategy for improving health outcomes in this population.

The purpose of this study was to understand the epidemiological and biological character-istics of Clostridium perfringens in avian origin in Yangzhou.A total of 200 intestinal samples(90 from chickens and 110 from ducks)collected from different markets were isolated and identified by isolation and culture,morphological observation,staining microscopy,biochemical tests and other methods,antimicrobial susceptibility test by K-B paper method,and toxin type of isolated bacteria was detected by PCR.Five isolates were screened according to the characteristics of host origin,se-rotype and drug resistance,and multi-locus sequence typing analysis was carried out,and a phylo-genetic tree based on α toxin gene sequences was constructed for isolates from different sources.After morphological and molecular biological identification,a total of 61 strains of Clostridium perfringens were isolated,and the isolation rate was 30.5％,all of which were type A.The antimi-crobial susceptibility results showed that the isolate had the highest resistance rate to amikacin,followed by tetracycline and cotrimoxazole,and was more sensitive to ceftriaxone,penicillin,amoxicillin and imipenem.Multi-drug resistance is very serious,with six or more strains resistant to antibiotics emerging in every market.A total of five different ST types(STs)were found by multi-site sequence analysis,namely ST833,ST834,ST363,ST835,and ST837,and all of them were new ST types except ST363.The information from ST 363 is consistent with clinical isolate of Clostridium perfringens,suggesting that more attention needs to be paid to zoonotic diseases caused by Clostridium perfringens.The α toxin genes of the five isolates were located in four dif-ferent branches in the phylogenetic tree,indicating their genetic diversity,while the isolates from different provenances of the same source were located in the same branch,indicating that they had a closer evolutionary relationship.The Clostridium perfringens from poultry in Yangzhou are mainly type A,and with severe drug resistance and multi-drug resistance present.This research provides scientific evidence for the epidemiology and control of Clostridium perfringens.

Objective:To investigate the role of telomere length in the causal effects of immune-mediated diseases on liver fibrosis.Methods:Genome-wide association study (GWAS) data were extracted from open GWAS (https: //gwas.mrcieu.ac.uk) for a two-sample Mendelian randomization (MR) analysis. Five immune-mediated autoimmune diseases (rheumatoid arthritis, systemic lupus erythematosus, primary biliary cirrhosis, primary biliary cholangitis, and Crohn′s disease) individually and collectively were included as exposure factors, telomere length as a mediator, and liver fibrosis as the outcome. The Wald ratio and inverse variance weighted (IVW) methods were performed to assess causal effects. The MR-Egger intercept test was adopted to evaluate the level of horizontal pleiotropy. Multivariable MR was employed to quantify the proportion of the effect of immune-mediated diseases on liver fibrosis mediated by telomere length. And sensitivity analyses were performed to assess the robustness of the results.Results:The results of IVW analysis revealed that the overall category of immune-mediated diseases, rheumatoid arthritis, systemic lupus erythematosus, primary biliary cirrhosis, primary biliary cholangitis, and Crohn′s disease were causally related to the high risk of liver fibrosis, and the OR were 1.63 (95% confidence intervals (95% CI): 1.33 to 2.10), 1.28 (95% CI: 1.14 to 1.43), 1.34 (95% CI: 1.02 to 1.74), 1.36 (95% CI: 1.27 to 1.47), 1.37 (95% CI: 1.23 to 1.52), and 1.52 (95% CI: 1.15 to 2.01), respectively ( P<0.001, <0.001, =0.032, <0.001, <0.001, =0.003). Horizontal pleiotropy was detected in the association between Crohn′s disease and liver fibrosis (MR-Egger intercept test, P=0.025).The results of multivariable MR indicated that telomere length acted as a mediating factor in the causal relationship between liver fibrosis and the overall category of immune-mediated diseases, rheumatoid arthritis, systemic lupus erythematosus, primary biliary cholangitis ( OR=2.24, 95% CI: 1.41 to 3.56; OR=1.78, 95% CI: 1.03 to 3.06; OR=2.11, 95% CI: 1.31 to 3.40; OR=2.01, 95% CI: 1.06 to 3.80; P<0.001, =0.038, =0.002, =0.032, respectively ). Conclusion:The causal effects of the overall category of immune-mediated diseases, rheumatoid arthritis, systemic lupus erythematosus, and primary biliary cholangitis on liver fibrosis are mediated by telomere length.

Diabetic kidney disease （DKD） stands as one of the most prevalent microvascular complications of diabetes，noted for its concealed onset and tendency to evolve into end-stage renal disease，profoundly impacting patients' life expectancy and quality of life. Epithelial-to-mesenchymal transition （EMT） is a central pathological process in the initiation and progression of DKD，facilitating disease advancement and renal fibrosis，thus representing a crucial focus of research into the pathological mechanisms of DKD. EMT is driven by the abnormal activation of signaling pathways，including transforming growth factor-β （TGF-β）/Smad，secreted glycoprotein/β-catenin，Notch，tumor necrosis factor-α （TNF-α）/nuclear factor-κB （NF-κB），and phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR），leading to renal cellular injury and subsequently accelerating renal fibrosis and the progression of DKD. Traditional Chinese medicine （TCM），characterized by its multi-target and multi-pathway therapeutic approach，demonstrates unique advantages in addressing DKD and EMT. Recent research has shown that active ingredients in TCM，including glycosides，flavonoids，and polyphenols，as well as TCM formulas，can precisely target these relevant signaling pathways，effectively inhibiting cellular injury in DKD and intervening in the EMT process. These findings not only underscore the potential of TCM monomers and formulas in treating DKD and EMT but also pave new directions for research in this field within TCM. This paper systematically reviewed the signaling pathways associated with EMT and provided an in-depth analysis of the research achievements and underlying mechanisms of TCM monomers and formulas in treating DKD and intervening in EMT，aiming to offer new insights and directions for TCM in the treatment of DKD and research on EMT，thereby further promoting the modernization and development of TCM.