Idiopathic Pulmonary Fibrosis (IPF) is characterized primarily by progressive lung tissue scarring. This study aimed to explore the molecular mechanism of the HOXA11-AS/miR-148a-3p/SMAD2 axis in IPF. Sixty IPF patients and 70 healthy controls were recruited. mRNA expression was measured by RT-qPCR. An IPF model was established in A549 cells and MRC-5 cells using transforming growth factor-beta 1 (TGF-β1) induction. Cell proliferation, apoptosis, oxidative stress levels, and fibrosis markers were assessed using the cell counting kit-8 (CCK-8), flow cytometry, and enzyme-linked immunosorbent assay (ELISA), respectively. A dual-luciferase reporter assay was conducted to confirm the regulatory interaction. In IPF patients, HOXA11-AS expression was significantly upregulated. Its expression was negatively correlated with lung function parameters as well as with the 6-minute walk test (6MWT) distance. Conversely, it was positively associated with fibrosis-related serum markers, such as hyaluronic acid (HA) and laminin (LN). These findings suggested that HOXA11-AS may serve as a promising diagnostic biomarker for IPF. Moreover, HOXA11-AS promoted the proliferation, suppressed apoptosis, enhanced oxidative stress by reducing superoxide dismutase (SOD) activity and increasing malondialdehyde (MDA) levels, and upregulated the key fibrosis markers expression, including alpha-smooth muscle actin (α-SMA), Collagen I, and Fibronectin. HOXA11-AS directly binds to miR-148a-3p, which in turn targets and regulates SMAD2. Notably, inhibition of miR-148a-3p reversed the suppressive effects of HOXA11-AS knockdown on TGF-β1-induced proliferation, oxidative stress, and fibrotic phenotypes in A549 cells, while restoring SMAD2 expression. Consistent results were observed in MRC-5 fibroblasts: HOXA11-AS knockdown significantly attenuated TGF-β1-induced proliferation, promoted apoptosis, and decreased the expression of α-SMA, Collagen I, and Fibronectin. These effects were partially reversed by miR-148a-3p inhibition, confirming the functional role of the HOXA11-AS/miR-148a-3p/SMAD2 axis in lung fibroblasts. HOXA11-AS promoted the IPF progression by upregulating SMAD2 expression through the sponging of miR-148a-3p.

To evaluate the ocular hypotensive effect of the Rho-associated kinase (ROCK) inhibitor RKI-1447 in normotensive and dexamethasone (DEX)-induced ocular hypertension rats, while elucidating its molecular mechanisms involving cytoskeletal remodeling and extracellular matrix (ECM) metabolism via the RhoA/ROCK pathway in trabecular meshwork (TM) cells. In vivo: Intraocular pressure (IOP) dynamics were monitored in normotensive and DEX-induced ocular hypertension rats following topical RKI-1447 administration. Conjunctival hyperemia, systemic safety (body weight), and IOP recovery post-withdrawal were assessed. In vitro: Primary human trabecular meshwork (HTM) cells were used to observe the effects of RKI-1447 on morphology and migration. Immunofluorescence assessed cross-linked actin networks (CLANs), ECM proteins, and alpha smooth muscle actin (α-SMA) expression. RhoA activity and the phosphorylation of myosin light chain (MLC) and myosin phosphatase target subunit-1 (MYPT-1) were detected via Pull-down assays and Western blot. Topical RKI-1447 was associated with a dose-dependent trend in IOP reduction in both normotensive and DEX-induced ocular hypertension rats, with a peak effect observed at approximately 2hours and only mild, transient hyperemia. In HTM cells, RKI-1447 treatment was observed to induce reversible morphological changes and mitigate cell migration. Furthermore, descriptive assessments suggested that RKI-1447 appeared to disassemble DEX-induced cytoskeletal assemblies and attenuate the upregulation of collage type IV (COL IV), fibronectin (FN), laminin (LN), and α-SMA. Additionally, RKI-1447 appeared to modulate DEX-associated RhoA activation and demonstrated a trend toward downregulating MLC and MYPT-1 phosphorylation. Our data suggest that RKI-1447 is associated with reduced aqueous outflow resistance, potentially through its inhibitory effect on the RhoA/ROCK pathway and the subsequent mitigation of steroid-induced cytoskeletal and fibrotic alterations. These preliminary observations suggest that RKI-1447 may serve as a viable pharmacological approach for treating steroid-induced glaucoma, although further long-term safety and efficacy evaluations are warranted.

The therapeutic efficacy of Gandouling (GDL) against hepatic fibrosis, along with its underlying mechanisms, was evaluated in a model of Wilson disease (WD). Using a copper-loaded rat model and in vitro LX-2 cell assays, we comprehensively evaluated the effects of GDL by employing transmission electron microscopy (TEM), histopathology, ultrasound elastography, confocal microscopy (mCherry-GFP-LC3 assay), qRT-PCR, and Western blotting. GDL treatment significantly improved liver function, indicated by reduced serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), hyaluronic acid (HA), laminin (LN), type III pre-collagen (PC-III), and collagen IV (C-IV). GDL also reduced the hepatic copper content and ameliorated histopathological and ultrasonographic signs of fibrosis. At the molecular level, GDL downregulated the expression of lncRNA SNHG7, DNMT3A, α-smooth muscle actin (α-SMA), and collagen I, while upregulating the expression of miR-29b. Furthermore, GDL inhibited autophagy, as shown by reduced levels of Beclin-1 and LC3-II/LC3-I and decreased autophagosome formation. These results demonstrate that GDL alleviates copper overload-induced hepatic fibrosis through modulation of the SNHG7/miR-29b/DNMT3A axis and inhibition of excessive autophagy.

This study aimed to investigate the effect and underlying mechanism of Taxus cuspidata seed oil (TCSO) on carbon tetrachloride (CCl4)-induced hepatic fibrosis in mice. A mouse model of hepatic fibrosis was established by CCl4 induction, and the model mice were subsequently treated orally with high dose or low dose TCSO for eight weeks. The degree of liver fibrosis and the mechanism of action were assessed through organ indices, serum biochemical markers, oxidative stress levels, histopathological examination, and molecular biological analyses. The results demonstrated that TCSO significantly reduced serum levels of alanine transaminase (ALT), aspartate transaminase (AST), and alkaline phosphatase (ALP). Concurrently, it decreased the concentrations of liver fibrosis markers, including procollagen III (PC III), collagen IV (IV-C), hyaluronic acid (HA), and laminin (LN), and reduced hepatic collagen deposition. Furthermore, TCSO enhanced the activities of the antioxidants superoxide dismutase (SOD) and glutathione (GSH) while inhibiting the production of the lipid peroxidation product malondialdehyde (MDA), and it ameliorated histopathological alterations in liver tissue. Additionally, TCSO markedly downregulated the expression of key fibrogenic proteins, such as transforming growth factor-β1 (TGF-β1), matrix metalloproteinase-2 (MMP-2), and tissue inhibitor of metalloproteinases-1 (TIMP-1), thereby effectively suppressing the progression of hepatic fibrosis. In conclusion, TCSO ameliorates hepatic fibrosis in mice by reducing hepatotoxic enzyme activity and collagen deposition, enhancing antioxidant capacity, and downregulating the expression of fibrosis-related proteins.

Three-dimensional (3D) models to study human disease mechanisms have demonstrated that the third dimension is an essential component for neuronal maturation and function. However, 3D neuronal cell culture is challenging due to their ultra-soft nature and specific extracellular matrix (ECM) organization. This study presents a microfiber reinforcement approach, combining primary mouse spinal cord neurons (SCNs) in a purpose-adaptable hyaluronic-acid-based matrix with melt electrowritten (MEW) frames to study disease mechanisms. The importance of laminins (LNs) is evaluated, which are vital for neuronal adhesion and maturation. Astrocytes (ACs) are mandatory in brain development and function by secretion of signal molecules, maintaining ion homeostasis, clearing of neurotransmitters, and by actively modulating neuronal activity. Three combinations are compared (i) isolated primary SCN, (ii) SCN with ACs (SCN-AC) and (iii) SCN-AC and LNs (SCN-AC-LN). Multimodal analysis by comparing protein expression, dendrite length, complex mechanical properties, and network functionality via Ca2+-imaging allows validation of structural and functional neuronal network development. The suitability of the 3D model to study pathomechanisms is demonstrated for Stiff Person Syndrome (SPS). For the first time, functional impairment of spinal cord neurons by patient-derived autoantibodies characteristic for SPS is recapitulated in a 3D spinal cord model platform adaptable to other disease types.

In this study, we describe the gelation kinetics, cytocompatibility, and mechanical properties of interpenetrating networks of collagen (COL), fibrin (FIB), hyaluronan (HA), and laminin (LAM) to evaluate their potential to produce mature skeletal muscle tissue. Skeletal muscle is a dynamic tissue that relies on the fusion of myoblasts into multinucleated myofibers to maintain homeostasis. In progressively degenerative conditions, impaired myoblast fusion leads to skeletal muscle atrophy and significant mass loss. Three-dimensional (3D) invitro models for skeletal muscle disease have been developed to better understand disease mechanisms and facilitate drug screening. However, most rely on Matrigel, a tumor-derived matrix that supports robust cell growth but has limited clinical relevance. To address this limitation, we focused on creating natural, multi-component scaffolds specifically tailored for muscle applications with clinically relevant drug testing use. Using spectrophotometry and rheology, we characterized the gelation kinetics and viscoelastic properties of interpenetrating networks with varying mass ratios of COL to FIB, supplemented with fixed proportions of HA and LAM. Tunable gelation was achieved within a range of 10 to 16 min. Cytocompatibility studies with C2C12 murine myoblasts demonstrated favorable cell viability in 1:1 and 1:2 (w/w) COL:FIB blends incorporating HA and LAM. Immunostaining of differentiated C2C12 cells confirmed Myosin 4 Monoclonal Antibody (MF-20) expression in these blends when seeded into polydimethylsiloxane (PDMS)-anchored bundles. Notably, in cell-laden 1:1 COL:FIB gels with a seeding density of 10 × 106 cells/mL, the compressive modulus increased three-fold between days 4 and 7 of differentiation. These findings highlight the potential of COL:FIB interpenetrating networks, enhanced with HA and LAM, as promising scaffolds for developing clinically relevant models of skeletal muscle tissue.

Therapeutic nanoparticle delivery is crucial for a variety of biomedical applications, such as immunizations, gene and drug delivery, tissue engineering, biomedical implant coating, and other regenerative medicine approaches. The uptake of therapeutic nanoparticles occurs through several endocytosis pathways. However, the uptake mechanism of nanoparticles delivered from a substrate is also regulated by cell-substrate interactions and the ability of cells to mechanosense their surrounding extracellular matrix (ECM). These cell-ECM interactions influence integrin signaling, focal adhesion formation, and cytoskeletal rearrangement to impact nanoparticle uptake. In this study, we investigated the role of ECM and ECM-mimetic coatings─collagen I (COL), fibronectin (FN), laminin (LM), hyaluronic acid (HA), and poly-l-lysine (PLL)─on the uptake of poly(lactic-co-glycolic acid) (PLGA) nanoparticles across three distinct cell types: NIH3T3 fibroblasts, primary rat adipose-derived stem cells (ASCs), and RAW264.7 macrophages, which displayed varying levels of integrin-based focal adhesion formation. Using a quartz crystal microbalance with dissipation (QCM-D) and ellipsometry, we thoroughly characterized ECM coatings, showing variations in coating thickness and mechanical properties. FN and COL coatings significantly enhanced cell proliferation, spreading, and focal adhesion formation, correlating with the highest levels of nanoparticle uptake at longer time points. In contrast, HA and LM coatings resulted in reduced cell adhesion and uptake. Consistent with this, cell types with more mature focal adhesions (ASCs, NIH3T3) showed much higher particle uptake in comparison to cells with limited focal adhesion formation (RAW264.7). Live-cell imaging demonstrated dynamic differences in uptake kinetics with LM coatings showing rapid early uptake, while FN and COL promoted sustained uptake over longer durations. Uptake studies using cytochalasin-D revealed that nanoparticle uptake was highly dependent on the actin cytoskeleton, suggesting the involvement of actin-dependent endocytic pathways. Overall, our findings highlight that ECM-dependent regulation of integrin-based adhesions and cytoskeletal organization modulates nanoparticle uptake in a coating- and cell type-dependent manner. These insights provide a foundation for optimizing substrate-based nanoparticle delivery platforms in regenerative medicine and therapeutic applications.

Fibroblast growth factor 19 in combination with liver fibrosis markers for non-invasive diagnosis of primary biliary cholangitis-associated cirrhosis.

EGCG inhibits hepatic stellate cell activity and liver fibrosis by targeting the MDM2/MUC5AC-mediated TGF-β1/Smad signaling pathway.

ObjectiveWilson disease (WD) is a rare autosomal recessive copper metabolism disorder, with hypersplenism as a severe, common complication secondary to disease-related cirrhosis. Currently, there is a lack of precise early prediction tools for this complication. This study aimed to construct a hypersplenism prediction model for WD patients by integrating multidimensional clinical indicators and machine learning, providing references for early identification of high-risk individuals and personalized interventions.MethodsA total of 524 WD patients were enrolled at the First Affiliated Hospital of Anhui University of Chinese Medicine from December 2019 to February 2025, including 244 with hypersplenism (HG) and 280 without (non-HG). After Key variables were selected through LASSO regression feature selection. Variate multicollinearity within the model was assessed using variance inflation factors (VIF). The predictive model was visualized using a nomogram. Five machine learning models were built with 10-fold cross-validation for parameter optimization. Finally, the model performance was evaluated, and the feature contributions were explained using the SHapley Additive exPlanations (SHAP) method.ResultsCompared with the non-HG group, the HG group had significantly lower WBC, PLT, and ceruloplasmin (CER), and higher A/G, PIIINP, CIV, hyaluronic acid (HA), laminin (LN), and 24-h urinary copper (CUU) (all p < 0.05). Multivariate logistic regression showed A/G, CIV, and PIIINP were independent risk factors, while WBC and PLT were independent protective factors. The SVM model performed best: training set AUC = 0.867 (95% CI: 0.830–0.904), accuracy = 0.807, specificity = 0.856, precision = 0.812, F1 score = 0.771; test set AUC = 0.771 (95% CI: 0.699–0.844) with AUC decay <10%. It also had excellent calibration (training set Brier score = 0.146, test set = 0.206) and clinical utility via DCA. SHAP analysis identified PIIINP as the core predictive feature, followed by WBC, PLT, and A/G, with CIV having relatively weaker influence.ConclusionThe SVM-based predictive model exhibits superior discriminatory power, calibration accuracy, and clinical utility for hypersplenism in WD patients. The five key features (WBC, PLT, A/G, CIV, PIIINP) with PIIINP as the core provide an objective quantitative basis for risk stratification, facilitating early identification and precise intervention of high-risk patients and improving WD prognosis.

Liver fibrosis after liver transplantation seriously affects graft survival. This study aimed to use shear wave elastography (SWE) to non-invasively evaluate the stiffness of the transplanted liver and spleen in liver transplant recipients. Seventy-one patients who were regularly reviewed in our hospital after liver transplantation (> 2years) were enrolled and were divided into two groups: Group B (30 cases without splenomegaly before liver transplantation) and Group C (41 cases with splenomegaly before liver transplantation). Besides, we selected 35 normal controls as Group A. All patients underwent conventional ultrasound, SWE, and serum liver fibrosis markers exams to obtain the oblique diameter of right lobe of liver (ODRL), portal vein diameter (PVD), portal vein velocity (PVV), peak systolic velocity (PSV), splenic vein diameter (SVD), splenic length, splenic thickness, liver stiffness measurement (LSM), spleen stiffness measurement (SSM) and laboratory test results. Research the correlation between SWE parameters and serum liver fibrosis markers by Pearson linear analysis. There was no difference in ODRL, PVD, PVV, and PSV among the three groups (all P > 0.05). Compared with Group A, hyaluronic acid (HA), laminin (LN), type III N-peptide collagen (PIIIP N-P), collagen type IV (IVC), and LSM were increased in Groups B and C (all P < 0.05) and were all higher in Group C than in Group B (all P < 0.05). Compared with Groups A and B, Group C was significantly higher in SVD, splenic length, splenic thickness, and SSM (all P < 0.05), whereas there was no difference in comparison of Group B with Group A (all P > 0.05). Pearson's correlation analysis showed that LSM and HA, LN, PIIIN-P, and IVC were positively correlated. SWE is valuable to evaluate the stiffness of transplanted liver and spleen in recipients after liver transplantation, and preoperative splenomegaly is a factor influencing graft fibrosis after transplantation.

ObjectiveSancai Lianmei granules (SCLMG) have demonstrated efficacy in improving glucolipid metabolism disorder, insulin resistance, and oxidative stress markers in both diabetic patients and rodent models. However, there is limited data available regarding the effect of SCLMG on human Metabolic dysfunction-associated fatty liver disease (MAFLD). This research aims to assess the impact of SCLMG on MAFLD in individuals with diabetes.Methods and research designSixty individuals diagnosed with type 2 diabetes mellitus (T2DM) and MAFLD were randomly allocated to one of two groups: the SCLMG group, which received standard treatment for T2DM along with SCLMG at a dose of 15 g three times daily, or the control group, which received standard treatment without SCLMG. The intervention lasted for 12 weeks. Alterations in liver fat content and liver sclerosis were evaluated using FibroScan. Secondary outcome measures included alterations in liver enzymes, fibrosis markers, advanced glycation end products AGEs, and metabolic parameters.ResultsWhen incorporated into the standard treatment regimen for MAFLD in diabetic patients, SCLMG exhibited an improvement in reducing hepatic steatosis (P = 0.048) while showing no substantial variations in liver stiffness (P = 0.762). Both the SCLMG and control groups revealed a substantial reduction in FibroScan readings at the end of the treatment period compared to baseline. The two groups exhibited substantial disparities in changes in liver enzymes (alanine aminotransferase (ALT) P = 0.018, aspartate aminotransferase (AST) P = 0.006, gamma-glutamyltranspeptidase (GGT) P < 0.001), skin autofluorescence (SAF) (P = 0.012), and metabolic parameters (P < 0.05). Nevertheless, there was a variation between the groups regarding serum procollagen III peptide (PIIIP) levels (P = 0.026), whereas changes in type Ⅳcollagen (CⅣ) (P = 0.265), hyaluronic acid (HA) (P = 0.199), laminin (LN) (P = 0.144), and high-density lipoprotein cholesterol (HDL-C) (P = 0.315) levels were not statistically significant.ConclusionsSCLMG is beneficial in regulating glucolipid metabolism and liver function. To a certain extent, SCLMG can improve liver steatosis and shows a tendency towards reducing liver sclerosis. Therefore, SCLMG has a good effect on individuals with T2DM and MAFLD.Trial registrationChiCTR2000033099||http://www.chictr.org.cn/ 20 May 2020.Registration details athttps://www.chictr.org.cn/hvshowprojectEN.html?id=34391&v=1.5Supplementary InformationThe online version contains supplementary material available at 10.1186/s12986-025-00996-z.

Kanglexin, a new anthraquinone compound, attenuates hepatic fibrosis by regulating the TGF-β/SMADS signaling pathway and glutathione metabolism.

To investigate the expression of the long non-coding RNA maternally expressed gene 3 (LncRNA Meg3) in patients with the Wilson disease (WD) and its correlation with the severity of liver fibrosis and autophagy-related markers. A total of 100 WD patients and 50 healthy individuals were enrolled from the First Affiliated Hospital of Anhui University of Chinese Medicine. Serum biomarkers, including platelet count, hyaluronic acid (HA), laminin (LN), type III procollagen N-terminal peptide (PIIINP), type IV collagen (C‑IV), alanine aminotransferase (ALT), and aspartate aminotransferase (AST), were measured, and the non-invasive indices APRI and FIB-4 were calculated. Peripheral blood levels of LncRNA Meg3, Beclin-1 and LC3B were detected using RT-qPCR, and liver stiffness (LSM) and shear wave velocity (SWV) were evaluated using two-dimensional shear wave elastography (2D-SWE). The liver tissues from 10 WD patients and 10 patients with hepatic hemangioma were examined using histochemical staining, transmission electron microscopy, and RT-qPCR. The expression level of LncRNA Meg3 was significantly lower, while the levels of AST, ALT, HA, LN, PIIINP, C‑IV, APRI, FIB-4, LSM and SWV were significantly higher in WD patients than in the healthy individuals (all P<0.01). LncRNA Meg3 was negatively correlated with LSM, SWV, APRI, FIB-4, Beclin-1 and LC3B (P<0.05). ROC analysis demonstrated that LncRNA Meg3 effectively discriminated >F4 stage fibrosis (AUC=0.902) with a sensitivity of 92.9% and a specificity of 83.7% at the optimal cut-off value, outperforming APRI (AUC=0.746) and FIB-4 (AUC=0.661). The liver tissues from WD patients exhibited characteristic histopathological changes and lowered expression of LncRNA Meg3, which was negatively correlated with Beclin-1 and LC3B expressions (P<0.05). Liver fibrosis staging (7 S4 cases and 3 S3 cases) was significantly associated with LSM and SWV levels (P<0.05). The expression level of LncRNA Meg3 is significantly decreased in WD patients, which is negatively correlated with the severity of liver fibrosis and closely related to the level of autophagy.

Objective: To investigate the potential therapeutic benefits of autoimmune antibody detection in individuals suffering from chronic hepatitis B. Methods: For the observation group, 102 patients with chronic hepatitis B who were admitted between March 2022 and March 2025 were chosen. Additionally, the control group consisted of 102 healthy people who were examined throughout the same time period. The two groups' autoimmune antibodies were identified, and patients in the observation group with positive and negative autoimmune antibodies were compared in terms of their liver function, liver fibrosis markers, and cytokine levels. Results: The total positive rate of autoimmune antibodies in the observation group was 26.47%, whereas it was 3.92% in the control group (P&lt;0.05). Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels were noticeably greater in the observation group's patients with positive autoimmune antibodies than in those with negative antibodies. However, the albumin (ALB) and total protein (TP) levels were much lower than those of patients with negative antibodies. A statistically significant difference was observed (P&lt;0.05). Type III procollagen (PCIII), hyaluronidase (HA), and the levels of laminin (LN) were statistically significant (P&lt;0.05) higher in individuals with positive autoimmune antibodies than in those with negative antibodies. Patients with positive autoimmune antibodies had considerably higher levels of interleukin-4 (IL-4), interleukin-10 (IL-10), and interleukin-6 (IL-6) than patients with negative antibodies. Patients with negative antibodies had significantly higher levels of interferon-γ (IFN-γ) (P &lt; 0.05). Conclusion: Autoimmune antibodies are present in patients with chronic hepatitis B, which influence liver fibrosis indicators and related cytokines. The identification of autoimmune antibodies in chronic hepatitis B patients can serve as a guide for evaluating the illness and determining prognosis.

This study aimed to investigate the association between the senescence-associated secretory phenotype (SASP) and balneotherapy (BT) in the elderly. Patients over the age of 65 with simple osteoarthritis were screened for their demographic characteristics, and those prone to inflammaging were included. Individuals were divided into two groups: the BT group (n = 20) and the control group (n = 20). Patients in both groups performed home-based relaxation and active range of motion (ROM) exercises three times a week. SASP levels were evaluated in both groups before and after 2 weeks of intervention. In the BT group, interleukin-8 (IL-8) (p = 0.044), laminin (LN) (p < 0.001), fibroblast growth factor-2 (p < 0.001), membrane cofactor protein (MCP-2) (p = 0.002), interleukin-1β (p = 0.002), metalloproteinase-3 (MMP-3) (p = 0.028), and tumor necrosis factor-α (TNF-α) (p = 0.044) increased following treatment, while MMP-1 (p = 0.019) decreased. In the control group, LN (p < 0.001), MMP-3 (p = 0.044), and MCP-2 (p < 0.001) increased, whereas TNF-α (p < 0.001) decreased. Intergroup comparisons showed significant changes in LN (p = 0.001), TNF-α (p < 0.001), and MMP-10 (p = 0.003). TNF-α and LN levels increased in favor of the BT group. A significant positive correlation was observed between TNF-α and the markers IL-8 (r = 0.344, p = 0.030), plasminogen activator inhibitor (r = 0.313, p = 0.050), LN (r = 0.328, p = 0.039), and glutathione (r = 0.374, p = 0.018). Our findings support the hypothesis that BT, through heat and chemical stressors, triggers a controlled inflammatory response that facilitates tissue regeneration while also mitigating oxidative stress to prevent excessive damage in aged individuals with potential inflammaging.

Fatty acid accumulation and lipid alterations play crucial roles in the proliferation and progression of breast cancer cells. In this study, we investigated molecular changes in triple-negative breast cancer (TNBC) tumors compared with matched normal breast tissues collected from three patients. Tumor and normal samples were analyzed using Raman microspectroscopy integrated with liquid chromatography-mass spectrometry (LC-MS) to identify ω-3, ω-6, and ω-9 fatty acids and key proteins, including collagen types I, III, IV, and V; fibronectin; and laminin. Additionally, proteomics and lipidomics analyses were conducted to characterize chemical and metabolic alterations between TNBC and normal breast adipose tissues. Raman spectral analysis showed a significant increase in unsaturated lipids in TNBC tissues compared to matched control tissues. The direct classical least-squares (DCLS) approach was used to analyze Raman spectra collected from entire tissues to identify unsaturated fatty acids (including ω-3 and ω-6) and proteins from both cancerous and matched normal breast tissues obtained from the same patients. Both Raman score data from DCLS analysis and LC-MS lipid analysis demonstrated an increase in the levels of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), arachidonic acid (AA), linoleic acid (LA), ω-3, and ω-6 and the ratio of LA/α-LA in tumors derived from TNBC compared to matched control tissues. Protein score data from DCLS analysis were consistent with our proteomics results for all proteins evaluated. The levels of collagen I (COL I), collagen III (COL III), collagen IV (COL IV), and laminin (LAM) were decreased in the TNBC tumor samples, while the levels of collagen V (COL V) and fibronectin (FN) were increased in the TNBC tumor samples. Lipidomics results showed that phosphatidylinositol (PI), phosphatidylethanolamine (PE), and phosphatidylglycerol (PG) lipid species were upregulated in TNBC. Network and pathway analyses have shown that coordinated changes in lipid metabolism and extracellular matrix (ECM) remodeling are key features of TNBC. This study designed here demonstrates the accuracy of Raman microspectroscopy as a nondestructive method for spatial profiling of lipid and protein expression on tumor slides. Further, due to the heterogeneity of breast cancer tumors and the role of the tumor microenvironment (TME) in breast cancer progression, spatial mapping enables mechanisms for profiling key extracellular components that may drive breast cancer progression and intratumor heterogeneity.

Background: Chronic liver disease has become a major health concern in people living with human immunodeficiency virus (PLWH). Clusterin has been involved in various pathologic conditions and its contributory role in liver complications in PLWH is still unclear. Objective: This study aimed to detect plasma clusterin levels and assess their correlation with liver fibrosis in Thai PLWH. Materials and methods: The study was conducted on 112 subjects, HIV-infected groups with (N=43) and without liver fibrosis (N=41), and uninfected controls (N=28). The subjects were evaluated for liver fibrosis using fibrosis-4 (FIB-4) score and aspartate aminotransferase to platelet ratio index (APRI), together with extracellular matrix fibrosis markers, laminin (LN), procollagen type III N-terminal peptide (PIIINP), hyaluronic acid (HA) and type IV collagen (IVC) and measured for plasma clusterin levels using an enzyme-linked immunosorbent assay. Results: This study reported that medians of clusterin levels in uninfected controls, and the HIV patient groups without and with liver fibrosis were 1,779.38 (373.07-5,578.69), 1,602.73 (445.28-3,738.33) and 1,487.17 (0-3,111.33) μg/mL, respectively. Kruskal-Wallis test demonstrated a downward trend of clusterin levels in the HIV-infected groups without liver fibrosis and a significantly decreased level in the group with liver fibrosis (p&lt;0.05), compared to those in uninfected controls. Pearson’s correlation analysis indicated negative correlations of clusterin levels with fibrosis markers, FIB-4 score (r=-0.224, p=0.018), APRI (r=-0.211, p=0.026), LN (r=-0.284, P=0.005), HA (r=-0.234, p=0.013) and IVC (r=-0.299, p=0.002). Conclusion: The study reported the lower levels of plasma clusterin in PLWH who develop liver fibrosis and their correlations with liver fibrosis assessed by the non-invasive markers.

The current study aims to elucidate the underlying pharmacological mechanisms of the Tibetan medicine Renqing Mangjue against hepatic fibrosis using a research strategy integrating network pharmacology and experimental verification. Through the combination of a rat hepatic fibrosis model induced by diethylnitrosamine(DEN) and the analysis of clinical transcriptome data from the GEO database, the characteristics of its pharmacological effects were objectively evaluated, and its mechanism of action was systematically analyzed. The experimental results showed that the intervention of Renqing Mangjue significantly reduced the levels of serum hepatic fibrosis markers, including procollagen type Ⅲ(PCⅢ), collagen type Ⅳ(Ⅳ-C), and laminin(LN). Specifically, the medium-dose group showed a decrease of 37.8%, 47.1%, and 36.6% respectively compared with the model group(all P &lt; 0.05) and a reduced collagen deposition in the liver tissue. The mechanism study found that it down-regulated the expressions of protein arginine methyltransferase 1(PRMT1), histone H4 arginine 3 dimethylation(H4R3me2a), plasminogen activator inhibitor-1(PAI-1), and monocyte chemoattractant protein 1(MCP-1) and inhibited the PRMT1/H4R3me2a/PAI-1/MCP-1 signaling axis, thereby inhibiting the activation of hepatic stellate cells and the excessive deposition of the extracellular matrix. This study is the first to clarify that Renqing Mangjue may exert an anti-hepatic fibrosis effect through the epigenetic regulation pathway, providing an experimental basis for its clinical application and offering a new paradigm for modern research on Tibetan medicine compounds.

ABSTRACT The crossing of the endothelial basement membranes (BMs) is a limiting step for leukocyte diapedesis. Heterotrimeric laminins (LMs) containing α4- and α5-chains are major BM components with opposite effects on transendothelial migration. Here, we examined whether LMα4 levels influence intratumor accumulation of specific immune cells and their impact on prognosis of early-stage colorectal cancer (CRC). Using two independent patient cohorts, we found that LMα4 expression positively correlated with intratumor infiltration of CD8+ T cells and macrophages, but not with regulatory T (Treg) cells. Kaplan-Meier and multivariate Cox regression analyses identified CD8+ T cell density as the strongest independent prognostic factor associated with reduced tumor relapse in both cohorts. While intratumor macrophage and Treg cell densities alone were not independently associated with prognosis, their abundance modulated outcomes specifically in tumors with high CD8+ T cell infiltration, with macrophage-rich tumors showing improved outcomes and Treg cell-enriched tumors exhibiting worse prognosis. Analysis of The Cancer Genome Atlas (TCGA) COAD cohort confirmed the positive correlation of LMα4 expression with both CD8+ T cell and macrophage infiltration, an association that was independent of the CRC clinical stage. Our findings suggest a subtype-specific effect for LMα4 in intratumor leukocyte infiltration, and underscore the prognostic interactions among CD8+ T cells, Treg cells and macrophages in early-stage colorectal cancer.