ObjectiveThis study systematically analyzed the arginine metabolic dysregulation in the rat model of heart failure （HF）， providing a modern scientific basis for elucidating the pathogenesis of HF and offering new insights for the prevention and treatment of HF with traditional Chinese medicine （TCM）. MethodsA thoracotomy was performed to ligate the left anterior descending coronary artery of rats， which induced acute myocardial ischemia and thus led to the development of post-myocardial infarction heart failure. The rats were divided into a sham surgery group and a model group， with eight rats in each group. Serum targeted metabolomics analysis was performed using ultra-performance liquid chromatography-triple quadrupole mass spectrometry （UPLC-TQ-S）， and the spatial distribution of metabolites in cardiac tissue was observed using airflow-assisted desorption electrospray ionizationmass spectrometry imaging （AFADESI-MSI）. Targets associated with HF and arginine metabolism were screened from databases including GeneCards and the Gene Expression Omnibus （GEO）， a protein-protein interaction （PPI） network was constructed， and enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway and Gene Ontology （GO） was performed. Finally， molecular docking was conducted to verify the binding between core metabolic components and key targets， and potential TCMs were predicted based on the core pathways and targets. ResultsCompared with the sham surgery group， the levels of arginine and citrulline in the serum of model rats were significantly decreased （P<0.01）， while those of proline， ornithine， creatine， creatinine and glutamate were significantly increased （P<0.05， P<0.01）. Cardiac mass spectrometry imaging showed a decreased abundance of arginine in the local myocardial tissue. Bioinformatics analysis identified 24 core functional targets， such as the angiotensin-converting enzyme （ACE）， neuronal nitric oxide synthase （NOS1）， 5-hydroxytryptamine receptor 2A （HTR2A）， and epidermal growth factor receptor （EGFR）， and enrichment analysis indicated that these targets were significantly involved in the calcium signaling pathway， neuroactive ligand-receptor interactions， and phosphatidylinositol signaling pathway. Molecular docking confirmed strong binding activities between arginine， citrulline and HTR2A， as well as between creatine， creatinine and EGFR. Based on pathway-target prediction， potential TCM interventions， such as ginseng and magnolia， were identified. ConclusionThis study revealed characteristic arginine metabolic disorder in HF， and the core targets of HF were closely associated with the phosphatidylinositol signaling pathway. It provides a modern biological interpretation of the pathogenesis of HF in TCM from the perspectives of metabolites and signaling pathways， and offers valuable insights for targeted therapy of HF and the development of TCM.

ObjectiveThis study systematically analyzed the arginine metabolic dysregulation in the rat model of heart failure （HF）， providing a modern scientific basis for elucidating the pathogenesis of HF and offering new insights for the prevention and treatment of HF with traditional Chinese medicine （TCM）. MethodsA thoracotomy was performed to ligate the left anterior descending coronary artery of rats， which induced acute myocardial ischemia and thus led to the development of post-myocardial infarction heart failure. The rats were divided into a sham surgery group and a model group， with eight rats in each group. Serum targeted metabolomics analysis was performed using ultra-performance liquid chromatography-triple quadrupole mass spectrometry （UPLC-TQ-S）， and the spatial distribution of metabolites in cardiac tissue was observed using airflow-assisted desorption electrospray ionizationmass spectrometry imaging （AFADESI-MSI）. Targets associated with HF and arginine metabolism were screened from databases including GeneCards and the Gene Expression Omnibus （GEO）， a protein-protein interaction （PPI） network was constructed， and enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway and Gene Ontology （GO） was performed. Finally， molecular docking was conducted to verify the binding between core metabolic components and key targets， and potential TCMs were predicted based on the core pathways and targets. ResultsCompared with the sham surgery group， the levels of arginine and citrulline in the serum of model rats were significantly decreased （P<0.01）， while those of proline， ornithine， creatine， creatinine and glutamate were significantly increased （P<0.05， P<0.01）. Cardiac mass spectrometry imaging showed a decreased abundance of arginine in the local myocardial tissue. Bioinformatics analysis identified 24 core functional targets， such as the angiotensin-converting enzyme （ACE）， neuronal nitric oxide synthase （NOS1）， 5-hydroxytryptamine receptor 2A （HTR2A）， and epidermal growth factor receptor （EGFR）， and enrichment analysis indicated that these targets were significantly involved in the calcium signaling pathway， neuroactive ligand-receptor interactions， and phosphatidylinositol signaling pathway. Molecular docking confirmed strong binding activities between arginine， citrulline and HTR2A， as well as between creatine， creatinine and EGFR. Based on pathway-target prediction， potential TCM interventions， such as ginseng and magnolia， were identified. ConclusionThis study revealed characteristic arginine metabolic disorder in HF， and the core targets of HF were closely associated with the phosphatidylinositol signaling pathway. It provides a modern biological interpretation of the pathogenesis of HF in TCM from the perspectives of metabolites and signaling pathways， and offers valuable insights for targeted therapy of HF and the development of TCM.

Objective To establish an epirubicin (EPI)-resistant murine triple-negative breast cancer (TNBC) (4T1/EPI) cell line and evaluate its biological characteristics and drug resistance. Methods The EPI-resistant cell line 4T1/EPI was developed through intermittent induction with gradually increasing EPI concentrations in vitro. Morphological changes were observed under an inverted microscope. Drug resistance index (MTT assay), cell doubling time (CCK-8 assay), and migration ability (wound healing assay) were evaluated. Western blot was used to detect the expression of drug resistance-related proteins. Transcriptome sequencing and KEGG pathway enrichment analysis were performed to identify the pathways and targets involved in EPI resistance, followed by experimental validation. Results The 4T1 cells eventually grew normally in a medium containing 100 ng/mL EPI, confirming the establishment of the 4T1/EPI resistant cell line. After stable resistance was acquired, morphological alterations were observed. Compared with their parental 4T1 cells, 4T1/EPI cells showed significantly prolonged doubling time (P<0.01) and enhanced migration ability (P<0.05). Expression levels of drug resistance-related proteins MDR1, MRP1 (P<0.01), and ABCG2 (P<0.05) were elevated in 4T1/EPI cells. In vivo models also demonstrated significant EPI resistance in 4T1/EPI tumors in terms of tumor weight and volume. Transcriptome sequencing highlighted the involvement of the PI3K/Akt signaling pathway and ABC transporter pathway. Validation experiments showed the upregulation of Erbb3, Egfr, PI3K, and Akt (P<0.05) and significant downregulation of Fgfr1 (P<0.01) in 4T1/EPI cells. Conclusion The EPI-resistant TNBC cell line 4T1/EPI was successfully established, exhibiting significant resistance in vitro and in vivo. The mechanism may involve the EPI-induced upregulation of Egfr and Erbb3, activating the PI3K/Akt pathway and subsequently enhancing ABC transporter expression.

This paper presents an automatic acquisition and analytic procedure of acupuncture manipulation based on optical navigation, aiming at solving the shortcomings of existing acquisition methods of acupuncture manipulation. An acquisition holder installed at the handle tail of filiform needle was designed to display the movement trajectory of the needle during acupuncture delivery by collecting the movement trajectory of holder. The 3-month old male Bama miniature pig was selected as the experimental subject, and 6 points, "Bojian" "Qiangfeng" "Housanli" "Xiaokua" "Huiyang" (BL35) and "Baihui" (GV20), were selected during acupuncture manipulation. The optical navigation system was used to collect the real-time data, and these data were per-processed and analyzed using mean filtering and Fourier transform. The acupuncture procedure was divided into 3 stages, inserting, lifting-thrusting, and twisting. The results showed that the accuracy was 96.3% at lifting-thrusting stage, and that was 100.0% at twisting stage. The decomposition effect of the entire procedure was satisfactory. This study provides a new approach to the quantitative analysis of acupuncture manipulation. In the future, it needs to further optimize the algorithm and expand the sample size so as to improve the accuracy of this analytic technique.
Acupuncture Therapy/methods* ; Male ; Animals ; Swine ; Acupuncture Points ; Humans ; Swine, Miniature ; Needles

Intestinal fibrosis is a major complication of inflammatory bowel disease (IBD), leading to a high incidence of surgical interventions and significant disability. Despite its clinical relevance, no targeted pharmacological therapies are currently available. This review aims to explore the underlying mechanisms driving intestinal fibrosis and address unresolved scientific questions, offering insights into potential future therapeutic strategies. We conducted a literature review using data from PubMed up to October 2024, focusing on studies related to IBD and fibrosis. Intestinal fibrosis results from a complex network involving stromal cells, immune cells, epithelial cells, and the gut microbiota. Chronic inflammation, driven by factors such as dysbiosis, epithelial injury, and immune activation, leads to the production of cytokines like interleukin (IL)-1β, IL-17, and transforming growth factor (TGF)-β. These mediators activate various stromal cell populations, including fibroblasts, pericytes, and smooth muscle cells. The activated stromal cells secrete excessive extracellular matrix components, thereby promoting fibrosis. Additionally, stromal cells influence the immune microenvironment through cytokine production. Future research would focus on elucidating the temporal and spatial relationships between immune cell-driven inflammation and stromal cell-mediated fibrosis. Additionally, investigations are needed to clarify the differentiation origins of excessive extracellular matrix-producing cells, particularly fibroblast activation protein (FAP) + fibroblasts, in the context of intestinal fibrosis. In conclusion, aberrant stromal cell activation, triggered by upstream immune signals, is a key mechanism underlying intestinal fibrosis. Further investigations into immune-stromal cell interactions and stromal cell activation are essential for the development of therapeutic strategies to prevent, alleviate, and potentially reverse fibrosis.
Humans ; Fibrosis/metabolism* ; Inflammatory Bowel Diseases/pathology* ; Animals ; Transforming Growth Factor beta/metabolism* ; Intestines/pathology*

Objective To investigate the safety and efficacy of hepatic arterial infusion chemotherapy(HAIC)using lenvatinib plus immune checkpoint inhibitors(ICI)in the treatment of progressive advanced hepatocellular carcinoma(HCC).Methods The clinical data of 67 patients with HCC of BCLC stage C,who received HAIC treatment at the Guangdong Provincial Hospital of Traditional Chinese Medicine of China from May 2018 to March 2022 and were assessed as in disease progression(PD)status after HAIC treatment,then had to further receive second-line treatment with lenvatinib plus ICI regimen.According to modified Response Evaluation Criteria in Solid Tumors(mRECIST),the tumor responses,including overall survival(OS),progression free survival(PFS),objective response rate(ORR),and disease control rate(DCR),were evaluated.Univariate analysis and multivariate analysis based on the COX proportional hazards regression model were used to determine the prognosis-related risk factors.According to the Common Terminology Criteria for Adverse Events version 5.0(CTCAE 5.0),the treatment-related adverse effects were recorded and evaluated.Results The median OS and median PFS in the 67 patients with HCC of BCLC stage C receiving second-line treatment with lenvatinib plus ICI regimen,who were evaluated as in PD status after first-line HAIC treatment,were 18.4 months and 7.2 months respectively.The ORR and DCR were 22.4％(15/67)and 67.4％(45/67)respectively.Univariate analysis and multivariate analysis showed that Eastern Cooperative oncology Group(ECOG)score and hepatitis B virus(HBV)infection were the independent risk factors affecting OS,while AFP level was an independent risk factor affecting PFS.The main treatment-related adverse reactions included elevation of transaminase level,hypertension,and hyperbilirubinemia.No death occurred during the therapeutic course.Conclusion For patients with HCC of BCLC stage C,who become in PD status after first-line HAIC treatment,second-line treatment with lenvatinib plus ICI regimen carries satisfactory efficacy and its adverse reaction is tolerable.

Objective : To investigate the impact of fecal microbiota transplantation (FMT) on the composition of 15 intestinal-derived estrogens and their metabolites (EMs) and its correlation with non-alcoholic fatty liver disease (NAFLD) . Methods: Thirty male C57BL/6J mice were divided into a normal control group (Control) , a high- sugar high-fat diet combined with low-dose CCl4 -induced NAFLD model group ( Model) , and a group of model mice treated with fecal microbiota from normal female mice (FMT) . After 17 weeks of modeling , liver pathology in each group was observed using HE staining , biochemical methods were used to measure serum alanine aminotrans- ferase (ALT) and aspartate aminotransferase (AST) levels , as well as hepatic triglyceride (TG) and total choles- terol (TC) levels. and the content of 15 EMs in portal vein serum was detected using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS) . The correlation between disease phenotype and intesti- nal EMs was analyzed using Pearson ′s method. Results: The NAFLD model was successfully established , and the FMT group showed improved liver structure and morphology , with significant decreases in liver function and hepatic lipids compared to the Model group. In NAFLD mice , the contents of E1 , E2 , and their 2- and 4-position metabo- lites in portal vein blood serum was reduced compared to normal mice , while the content of most 16- and 17-posi- tion metabolites ( except 16α-OHE1) increased compared to normal mice. Correlation analysis showed that ALT was strongly positively correlated with E3 and 17-epiE3 , and strongly negatively correlated with E1 , E2 , 4- MeOE1 , and 16α-OHE1 . The TC was strongly positively correlated with 17-epiE3 and strongly negatively correla- ted with E1 , 4-MeOE1 , and 16α-OHE1 . Conclusion FMT restores the disrupted composition of intestinal EMs and improves NAFLD.

In June 2018,sows at a pig farm in Jiamusi,Heilongjiang Province,suffered a large num-ber of miscarriages,and subsequently weaned piglets at the farm began to show persistent high fe-ver symptoms at around 35 days of age,with some pigs having a fever of more than 41.5 ℃.In or-der to determine the cause of this outbreak,63 clinical samples from this farm were tested.The re-sults showed that 60 out of 63 samples were positive for the porcine reproductive and respiratory syndrome virus(PRRSV)antigen.Subsequently,PRRSV antigen-positive plasmid was transfected into African green monkey embryonic kidney cells(Marc-145),and after three generations of blind transmission,indirect immunofluorescence assay(IFA)was performed.The results showed that one PRRSV strain,named HLJ38,was successfully isolated.Then the whole genome of HLJ38 strain was sequenced and then analyzed in detail by bioinformatics software.Sequence analysis showed that there were three deletions of 131 amino acids(323-433 aa,483 aa and 504-522 aa)in the derived sequence of Nsp2 gene of HLJ38 strain,which was consistent with the molecular ge-netic marker of NADC30 PRRSV.The phylogenetic tree analysis showed that HLJ38 and NADC30 PRRSV in GenBank belong to lineage 1 subgroup,and the nucleotide homology of HLJ38 and NADC30 PRRSV in GenBank was only 85.2％and 84.6％.Recombinant analysis showed that HLJ38 was a recombinant NADC30-like PRRSV,and the recombinant gene fragments were de-rived from multiple strains,among which the fragment of 1-201 nt was provided by VR2332 strain and fragment of 6 641-8 061 nt derived from the HP-PRRSV strain.In summary,the re-sults showed that the outbreak in this pig farm may be caused by the recombination of PRRSV strains among different lineages,and the recombinant circulating strains not only have certain pathogenicity but also suggest that the existing commercial vaccines provide limited cross-protec-tion against them.Recombination between different lineages increases the genetic diversity of PRRSV and aggravates the difficulty of prevention and control of PRRS in pig farms.Therefore,it is necessary to continuously monitor the epidemic dynamics of PRRSV in pig farms and take effec-tive measures in time to curb the spread of PRRS.

Objective To investigate the effects of acupoint application at Tianshu(ST25)combined with kidney-warming and spleen-invigorating therapy on clinical efficacy,intestinal function,and the expressions of Toll-like receptor 4(TLR4)and nuclear factor kappa-B(NF-κB)in patients with ulcerative colitis(UC).Methods A total of 100 UC patients with depletion and deficiency of spleen-kidney complicated with internal accumulation of pathogenic dampness syndrome,treated in the Department of Proctology,Shanghai Municipal Hospital of Traditional Chinese Medicine Affilated to Shanghai University of Traditional Chinese Medicine from June 2022 to June 2024,were selected and randomly divided into four groups(Groups A,B,C,D)using a random number table method,with 25 patients in each group.Group A received standard mesalazine treatment.Group B received Kidney-Warming and Spleen-Invigorating Formula(composed of Psoraleae Fructus,Myristicae Semen,Euodiae Fructus,Schisandrae Chinensis Fructus,Codonopsis Radix,Poria,Atractylodis Macrocephalae Rhizoma,etc.)combined with standard mesalazine treatment.Group C received acupoint application at Tianshu(the medicinal paste for application composed of Caryophylli Flos,Cinnamomi Cortex,Atractylodis Macrocephalae Rhizoma,Euodiae Fructus,Coptidis Rhizoma,etc.)combined with standard mesalazine treatment.Group D received Kidney-Warming and Spleen-Invigorating Formula+acupoint application at Tianshu+standard mesalazine treatment.All groups were treated continuously for 12 weeks.The time for disappearance of clinical symptoms was compared among the four groups.Serum immune cytokines[β-defensin-1,total immunoglobulin A(IgA),interleukin-6(IL-6),tumor necrosis factor-alpha(TNF-α)],peripheral blood levels of TLR4 and NF-κB,protein expression levels of TLR4 mRNA and NF-κB p65 mRNA in intestinal mucosal tissue,and changes in intestinal microbiota were observed before and after treatment.The clinical efficacy of the four groups was evaluated.Results(1)Compared to the other three groups,Group D had the shortest time for disappearance of clinical symptoms such as abdominal pain,diarrhea,mucopurulent bloody stools,and tenesmus(P＜0.05).The time for disappearance of various clinical symptoms in Groups B and C was similar(P＞0.05)but was shorter than that in Group A(P＜0.05).(2)After treatment,the serum levels of β-defensin-1,IgA,IL-6,and TNF-α in all four groups were significantly lower than those before treatment(P＜0.05).Post-treatment intergroup comparisons showed statistically significant differences(P＜0.01),with Group D exhibiting the greatest reduction,significantly lower than the other three groups(P＜0.05).The levels in Groups B and C were similar(P＞0.05)but were lower than those in Group A(P＜0.05).(3)After treatment,the peripheral blood levels of TLR4 and NF-κB,and the protein expression levels of TLR4 mRNA and NF-κB p65 mRNA in intestinal mucosal tissue showed a decreasing trend in all four groups(P＜0.05).Post-treatment intergroup comparison results were statistically significant(P＜0.01),with Group D showing the greatest reduction,significantly lower than the other three groups(P＜0.05).All above levels in Groups B and C were similar(P＞0.05)but were lower than those in Group A(P＜0.05).(4)After treatment,the relative contents of Bifidobacterium and Lactobacillus increased(P＜0.05),while those of Saccharomyces and Clostridium decreased(P＜0.05)in Groups B,C,and D.Post-treatment intergroup comparison results were statistically significant(P＜0.01).Group D showed a significant increase in Bifidobacterium and Lactobacillus and a significant decrease in Saccharomyces and Clostridium,with the magnitude of change significantly greater than that in the other three groups(P＜0.05).The quantities of these bacteria in Groups B and C were similar(P＞0.05),but the improvement was significantly greater than that in Group A(P＜0.05).(5)After 12 weeks of treatment,Group D had the highest total effective rate at 100.00％(25/25).The total effective rates of Groups B and C were similar,both at 84.00％(21/25),but were higher than that of Group A(56.00％,14/25).The intergroup difference was statistically significant(x2=16.310,P＜0.01).Conclusion The comprehensive treatment regimen of acupoint application at Tianshu combined with Kidney-Warming and Spleen-Invigorating Formula,on the basis of standard mesalazine treatment,has a positive impact on UC patients with depletion and deficiency of spleen-kidney complicated with internal accumulation of pathogenic dampness syndrome.This treatment method can effectively improve clinical symptoms,regulate β-defensin-1 and IgA levels,enhance immune defense function,downregulate the expression levels of TLR4 and NF-κB,and alleviate inflammatory response.Compared to monotherapies,it is more effective on improving treatment outcomes.