Background The simultaneous presence of sepsis and type 2 diabetic mellitus (T2DM) has a synergistic effect on hepatic damage. Although the activation of peroxisome proliferator-activate receptor gamma (PPARG) and AKT serine/threonine kinase 1 (AKT1) have been shown to confer protection alone in isolated models of sepsis or T2DM. The present study was designed to systematically assess the effects of PPARG or AKT1 activation on inflammatory responses and glucose and lipid metabolism as well as intestinal barrier function in a murine model of T2DM and sepsis (T2DM/sepsis), and understand the molecular pathways involved.Methods Mice were treated with either the PPARG agonist pioglitazone or the AKT1 activator SC79 and we then evaluated survival, inflammatory indicators, metabolic changes, tissue pathology, intestinal barrier integrity, and gut microbiota composition. Mechanistic studies focused on the PPARG/AMPK and PI3K/AKT1/mTOR signaling pathways.Results Activation of either PPARG or AKT1 was associated with significantly increased survival rates, reduced systemic and hepatic levels of pro-inflammatory cytokines, increased the expression of anti-inflammatory mediators, ameliorated the abnormalities of the glucose and lipid metabolism, and partially improved hepatic, pulmonary, and intestinal injury. Mechanistically, PPARG activation mainly activated the PPARG/AMPK pathway and AKT1 activation enhanced the PI3K/AKT1/mTOR pathway. Furthermore, the intestinal barrier integrity and gut microbiota composition was partially improved.Conclusion These results suggest that targeting PPARG or AKT1 is associated with protection against sepsis-associated liver injury in mice with T2DM by modulating the PPARG/AMPK axis or the PI3K/AKT1/mTOR pathway, respectively, which may collectively contribute to anti-inflammatogenic, metabolic, and gut-barrier protective mechanisms.

BACKGROUND:A major pathological event in heart failure (HF) is cardiac fibrosis induced by pressure overload, and infiltration of monocytes is essential in this scenario. Nevertheless, there are no targeted therapies. Ginsenoside Rb1 (Rb1), a major active component of Panax ginseng, has been proven to have cardioprotective ability, although the exact molecular target is not yet clearly understood. METHODS:Mouse models of cardiac fibrosis were induced by subcutaneous isoproterenol (ISO) injection or transverse aortic constriction (TAC). Echocardiography, histology, and molecular biology techniques were used to evaluate the effects of Rb1 on cardiac function, hypertrophy, fibrosis, and macrophage infiltration. The direct target of Rb1 was identified through a combined methodology that includes phosphoproteomics, thermal proteome profiling, cellular thermal shift assay, and surface plasmon resonance. The specific role of monocytic Raf1 was further validated using a pharmacological inhibitor (GW5074) and monocyte/macrophage-specific Raf1 knockdown in vivo. RESULTS:Rb1 was effective in the prevention of cardiac dysfunction, hypertrophy, and fibrosis in both ISO- and TAC-induced mouse models. These effects were mediated through the inhibition of monocyte recruitment by directly targeting Raf1. Mechanistically, Rb1 interacts with Raf1, inhibits its kinase activity, and consequently suppresses the Raf1-MEK-Erk-Creb signaling, which resulted in decreased CD11b/CD18 expression and inhibited migration of monocytes. Importantly, the anti-fibrotic effects of Rb1 were recapitulated by the Raf1 inhibitor GW5074 and in vivo monocyte/macrophage-specific Raf1 knockdown. CONCLUSION:Our research not only unveils a new immunomodulatory effect of Ginsenoside Rb1 that targets monocytic Raf1 as a key mediator but also notes Raf1 inhibition as a promising therapeutic strategy of HF.

OBJECTIVES:Bismuth-containing quadruple therapy (BQT) is recommended for H. pylori rescue treatment, but many patients cope with pill burden and adverse events. High-dose dual therapy (HDDT) may be easier to tolerate. We compared HDDT with BQT in rescue therapy. MATERIALS AND METHODS:We followed PRISMA 2020 (PROSPERO CRD420251266898) and searched PubMed/MEDLINE, Embase, Cochrane CENTRAL, Web of Science Core Collection, CiNii Research, and Major Chinese Databases (CNKI, Wanfang Data, and VIP) through December 16, 2025. We pooled head-to-head rescue RCTs comparing HDDT (PPI/PCAB + Amoxicillin ≥ 3 g/Day, ≥ TID) with BQT using random-effects models. risk of bias was assessed with RoB 2 and certainty with GRADE. TSA assessed information size for the primary ITT outcome. RESULTS:Ten RCTs (n = 2407) showed comparable ITT eradication for HDDT vs. BQT (79.9% vs. 79.9%, RR 1.01, 95% CI 0.97-1.05; I2 = 3.9%). Any adverse events were less frequent with HDDT (122/1135, 10.7%) than BQT (334/1123, 29.7%). The corresponding meta-analysis (expressed as AE-free so that RR > 1 favors HDDT) showed RR 1.27, 95% CI 1.15-1.41. TSA suggested that the required information size for the primary ITT outcome was reached, making a clinically important (≥ 10%) relative difference in ITT eradication unlikely. CONCLUSIONS:In rescue patients without penicillin allergy, HDDT achieves eradication comparable to BQT while causing far fewer adverse events. HDDT is a practical rescue option when BQT is poorly tolerated, although most evidence comes from China and reporting of adherence and resistance is uneven.

Vasogenic cerebral edema is a severe complication of delayed thrombolysis for ischemic stroke, for which no pharmacological treatment exists. Anisodine hydrobromide (Ani), an alkaloid used clinically in China for vascular disorders, is investigated for its potential to mitigate this condition. Here we show that Ani treatment improves survival and neurological function in a mouse model of delayed rtPA-induced cerebral edema by preserving the integrity of the blood-brain barrier. Utilizing proteomics and microarray screening, we identify megakaryocyte-associated tyrosine kinase (Matk) as a direct target of Ani. We demonstrate that Ani binding stabilizes Matk, preventing its degradation and suppressing the activation of Src kinase. This inhibition consequently blocks the dual paracellular and transcellular leakage pathways that drive vasogenic edema. Our findings reveal the Matk-Src signaling axis as a therapeutic target and support Ani as a promising clinical candidate for preventing post-thrombolytic complications in stroke management.

In mammals, the postnatal increase in cardiac workload coincides with the loss of cardiomyocyte proliferative capacity, rendering adult cardiomyocytes permanently post-mitotic. Whether reducing load can restore regenerative potential in the adult heart remains unknown. Here we use a heterotopic heart transplantation model, in which the donor heart is vascularized but nonpumping, to show that mechanical unloading induces adult cardiomyocyte proliferation, revealed by Ki67-based and MADM (mosaic analysis with double markers) lineage tracing. Applying heterotopic transplantation to infarcted hearts to achieve mechanical unloading similarly promoted regeneration within peri-infarct regions. Single-nucleus RNA sequencing of unloaded hearts identified enhanced epicardial-cardiomyocyte communication via the NRG1-ERBB4-STAT3 axis. Epicardial Nrg1 deletion blocked STAT3 activation and cardiomyocyte proliferation. CUT&Tag revealed STAT3 directly upregulates H6pd in cardiomyocytes, boosting pentose phosphate pathway activity to supply nucleotides and reducing equivalents for proliferation. These findings delineate a mechanotransductive pathway linking epicardial signals to cardiomyocyte cell-cycle re-entry, providing a framework for leveraging unloading to promote cardiac regeneration.

OBJECTIVE:Limb frostbite caused by cold exposure is a prevalent injury in winter. However, effective therapies remain limited, particularly for cold-induced edema. This study evaluated the therapeutic potential of DangGuiSiNi Decoction (DSD), QiShenYiQi Pills (QSYQ), and their combination in a rat model of cold-induced limb injury. METHODS:Male Sprague-Dawley rats (180-220 g) were subjected to hind limb frostbite through 5-min dry ice exposure. Animals were administered with DSD, QSYQ, or their combination via oral gavage either 2 h pre-exposure or 5 min post-exposure, followed by daily dosing for 7 days. Hennepin Score, hind limb blood flow, FITC-albumin extravasation, rhodamine 6G-labeled leukocyte adhesion, expression of tight/adhesion junction proteins and basement membrane components, transient receptor potential (TRP) channel proteins, ATP content, and norepinephrine levels at 2 h, 24 h, and 7 days post-exposure were assessed. RESULTS:The combination of DSD and QSYQ significantly alleviated frostbite-induced edema and gangrene. Compared with control group, cold exposure increased the expression of TRPA1, TRPM8, RhoA, ROCK, p-MLC, adrenergic receptor α2c and norepinephrine content, while decreased the expression of tight and adherent junction proteins claudin-5, ZO-1, JAM-1, basement membrane proteins laminin and collagen-IV, ATP content, and the activity of mitochondrial complex I, II, IV and V. DSD primarily counteracted the cold-sensing and vasoconstrictive pathways by reducing TRPA1/TRPM8 expression, norepinephrine levels, and RhoA/ROCK activation, thereby improving blood flow. In contrast, QSYQ enhanced vascular barrier integrity by upregulating junction and basement membrane proteins, and boosted energy supply by increasing ATP content and mitochondrial complex activities. The combination of DSD and QSYQ produced aggregated therapeutic effects. CONCLUSION:DSD attenuates frostbite by inhibiting temperature-sensing and vasoconstrictive pathways to ameliorate ischemia, whereas QSYQ protects vascular barrier function and mitigates edema by restoring energy metabolism. Their combination exhibits complementary and synergistic effects, supporting its use as a promising adjunctive therapy for cold-induced injury.

INTRODUCTION:Administration of recombinant tissue plasminogen activator (rtPA) beyond 4.5  h after ischemic stroke exacerbates blood-brain barrier (BBB) disruption, leading to vasogenic cerebral edema and hemorrhage. However, current therapies remain ineffective. OBJECTIVE:This study aimed to develop and validate an optimized multicomponent combination, termed ADR, consisting of Astragaloside IV (ASIV), 3,4-dihydroxyphenyl lactic acid (DLA), and Notoginsenoside R1 (R1), to prevent BBB damage following rtPA thrombolysis at 4.5  h after stroke onset in mice and to explore the underlying mechanisms. METHODS:ADR was optimized using a uniform design-entropy weight-regression model. Its efficacy was assessed in mice receiving rtPA at 4.5  h after stroke onset. The pharmacokinetic (PK) and pharmacodynamic (PD) properties of the components were evaluated. Multi-omics analysis, molecular docking, surface plasmon resonance (SPR), and cellular thermal shift assays (CETSA) were performed to identify key targets, followed by functional validation through gene silencing or overexpression in vitro and in vivo. RESULTS:The optimized ADR improved cerebral blood flow, reduced infarct size and neuronal apoptosis, ameliorated neurological deficits, and enhanced survival rates. It effectively inhibited microvascular leakage, hemorrhage, and leukocyte adhesion. PK and PD studies, along with in vivo pharmacological evaluation of the individual components, demonstrated that the combination produced synergistic effects. Integrated analyses identified five key molecules. Molecular docking, SPR, and CETSA confirmed that LMO7 was exclusively modulated by ADR, whereas CAPG was regulated by both ADR and all three components. Additionally, MOBP, HMGB2, and TAGLN2 were targeted by ASIV, DLA and R1, respectively. These findings were further confirmed by silencing LMO7 or overexpressing CAPG, MOBP, HMGB2, or TAGLN2 in vitro and in vivo. CONCLUSION:Our study demonstrated that ADR prevented BBB disruption following rtPA thrombolysis in mice with ischemic stroke through multitarget regulation and provided valuable insights into the integration of Traditional Chinese Medicine and modern pharmacology.

BACKGROUND:Pulmonary microcirculatory dysfunction is a hallmark of sepsis, contributing to hypoxemia, pulmonary edema, and multiple organ failure. Anisodamine hydrobromide (ADM), a natural alkaloid with anti-inflammatory and endothelial-protective properties, has been used clinically in China for septic shock. However, its effects on pulmonary microcirculatory dysfunction in septic shock remain unclear. METHODS:A rat model of sepsis was established via cecal ligation and puncture (CLP). Rats were treated with low, medium, or high doses of ADM. Seven-day survival rates and arterial blood gas parameters were monitored. Pulmonary microvascular leakage was evaluated using Evans blue extravasation and FITC-dextran imaging. Histological analysis, immunofluorescence, and Western blotting were performed to assess leukocyte adhesion, inflammatory cell infiltration, endothelial junction proteins, basement membrane proteins, and matrix metalloproteinases. RESULTS:ADM treatment significantly improved 7-day survival and restored arterial partial pressure of oxygen (PaO2), oxygen saturation (SaO2), and pH in CLP rats. High-dose ADM markedly reduced Evans blue and FITC-dextran leakage, attenuated pulmonary edema, and preserved alveolar architecture. ADM inhibited leukocyte adhesion in pulmonary microvessels and decreased infiltration of MPO- and CD68-positive inflammatory cells. Mechanistically, ADM suppressed CLP-induced Caveolin-1 upregulation, restored the expression of VE-Cadherin, Occludin, and Claudin-5, and prevented degradation of Collagen IV and Laminin. Additionally, ADM significantly downregulated MMP-9 expression, while MMP-2 levels remained unchanged, suggesting a role in limiting junctional and basement membrane degradation. CONCLUSION:ADM protects against CLP-induced pulmonary microcirculation dysfunction in rats by attenuating inflammatory cell infiltration, preserving pulmonary endothelial junctions, and maintaining basement membrane integrity. These results provide mechanistic insight into ADM's therapeutic potential in sepsis-induced pulmonary microcirculation dysfunction and support its use for sepsis in clinic.

ETHNOPHARMACOLOGICAL RELEVANCE:Zhi Chuan Ling Oral Liquid (ZCL) has long been used in traditional Chinese medicine for the treatment of asthma. However, how ZCL acts on Th2-type asthma and its active components remain unclear. AIM OF THE STUDY:This study is to investigate the mechanism and potential active substances of ZCL in intervening Th2-type asthma using an ovalbumin (OVA)-induced Th2-type asthma model. MATERIALS AND METHODS:An OVA-sensitized and challenged Th2-type asthma mouse model was established to evaluate the effects of ZCL on airway hyperresponsiveness, Th2-type inflammatory effector cells and cytokines in bronchoalveolar lavage fluid (BALF). Proteomics was employed to identify potential targets of ZCL intervention in Th2-type asthma, and validation was performed in vitro and in vivo using molecular biology, bioinformatics, and LC-MS. RESULTS:ZCL alleviated airway hyperresponsiveness and airway inflammation in Th2-type asthmatic mice, and downregulated TGF-β mRNA expression in lung tissue, decreased the numbers of eosinophils, mast cells and Th2 cytokines in BALF. Proteomic analysis suggested that ZCL may modulate Th2-type asthma via IL-4 and KIT signaling pathways. Western blotting showed that ZCL reduced the IL-4's downstream p-JAK1/JAK1 and p-STAT6/STAT6 ratios and decreased the expression of KIT and Tryptase in lung tissue. Bioinformatics analysis, LC-MS, and in vitro experiments suggest that rutin, forsythiaside, and isoforsythiaside may be the key components of ZCL responsible for intervening in the aforementioned targets. CONCLUSIONS:ZCL can ameliorate Th2-type asthma by suppressing IL-4 and KIT signaling pathways. Rutin, isoforsythiaside and forsythiaside are likely to be key bioactive components mediating these protective effects.

BACKGROUND:Acute lung injury (ALI) rapidly progressing into acute respiratory distress syndrome (ARDS) is a major cause of the high fatality rate in acute respiratory infectious diseases. Qing-Fei-Pai-Du-Tang (QFPDT) has a clinical advantage in ALI/ARDS treatment. However, whether QFPDT can improve multiple pathological process involved in pulmonary microcirculatory disturbances during ALI, and the underlying mechanisms remain unclear. PURPOSE:The present study aimed to assess the role of QFPDT in a lipopolysaccharide (LPS)-induced ALI rat model, attempting to disclose the rationale behind the effects of QFPDT. METHODS:Male Wistar rats were intraperitoneally injected with LPS (7.5 mg/kg), and received QFPDT (6 g/kg) by gavage either 10 min before (pre-treatment) or 6 hours after (post-treatment) LPS injection. Intravital microscopy, histology, immunohistochemistry and immunofluorescence, flow cytometry, enzyme-linked immunosorbent assay, Western blotting, and proteomics analysis were utilized to investigate the effects and mechanisms of QFPDT. Chemical profiling of QFPDT was performed to identify potential active ingredients. RESULTS:The results revealed that 6 and 24 hours after LPS injection induced a hyper-inflammatory and hyperpermeability process in rat lung tissues. Pre- and post-treatment with QFPDT attenuated the increase in leukocyte adhesion to pulmonary venules, accompanied by high expression of CD11b and intercellular adhesion molecule-1. Besides, QFPDT attenuated the LPS-induced increase in fluorescein isothiocyanate-dextran leakage from pulmonary microvessels, along with a downregulated expression of junction proteins and an upregulated expression and phosphorylation of Caveolin-1. Moreover, there was a downregulated expression of basement membrane proteins, increased matrix metalloproteinase-9 and cleaved Cathepsin B, and decreased ATP/ADP and ATP/AMP ratios after LPS, all of which were attenuated by QFPDT. Proteomics data evaluated by gene set enrichment analysis, QFPDT pharmacokinetic analysis, combined with molecular docking prediction and surface plasmon resonance validation revealed that QFPDT contained lung-entering prototype ingredients that improved ALI by regulating various key signaling pathway proteins associated with leukocyte adhesion, microvascular hyperpermeability, basement membrane degradation, and oxidative stress. CONCLUSION:The present study demonstrates the multifaceted effects of QFPDT and offers insight into better understanding its underlying mechanisms in attenuating LPS-induced pulmonary microcirculatory disturbances and lung dysfunction through a multi-component and multi-target mode, thereby providing evidence supporting the application of QFPDT in ALI/ARDS-related diseases.

Background:Immunoglobulin A nephropathy (IgAN) is one of the most common causes of primary glomerulonephritis that lacks a specific treatment option. This study aimed to evaluate the efficacy and safety of telitacicept in patients with IgAN. Methods:We performed a retrospective analysis in 82 biopsy-proven IgAN patients with baseline estimated glomerular filtration rate (eGFR) >20 mL/min/1.73 m2 and proteinuria ≥1 g/day. Forty-one patients were treated with telitacicept and angiotensin-converting enzyme inhibitor (ACEI)/angiotensin receptor blocker (ARB). They were divided into extended group (treated with telitacicept weekly for the first 6 months, then once every 2 weeks for the next 3-6 months) and short-term group (treated with telitacicept weekly for the first 6 months). The other 41 patients received ACEI/ARB alone and served as the ACEI/ARB group. Results:The mean percent change in proteinuria from baseline of extended group, short-term group and ACEI/ARB group were -56.8 ± 23.5% (P < .01), -28.6 ± 65.6% (P = .09) and -0.3 ± 57.0% at Month 12. eGFR decline in telitacicept groups were slower compared with the ACEI/ARB group. Univariate logistic regression analysis revealed only extended treatment (odds ratio = 4.3, 95% confidence interval 1.2-15.0, P < .05), but not short-term treatment was significantly associated with proteinuria decrease (defined as reduction in urine protein by more than 50%) at 12 months. This association remained robust after adjusting for age, gender, baseline eGFR or proteinuria. Subgroup analysis showed that the effect of extended treatment on reducing urine protein was more pronounced than that of short-term treatment in patients with higher proteinuria (≥2 g/day), poorer renal function (eGFR<60 mL/min/1.73 m2), or worse pathological changes (M1, E1, T1/T1 and C1/C2). The safety outcomes of telitacicept were similar to ACEI/ARB. No severe adverse events were reported in all groups. Conclusion:Our study confirms that telitacicept has a definite proteinuria-lowering effect in IgAN. Extending the treatment duration from 6 months to 9-12 months further enhances its ability to reduce proteinuria.

BACKGROUND:Acute lung injury (ALI) has emerged as a critical illness, with sepsis-related ALI accounting for >80 %. In the context of bacterial infection, damage to the pulmonary microvascular barrier leads to inflammatory cell infiltration and plasma component extravasation into pulmonary interstitium. This disruption impairs gas exchange, resulting in hypoxemia. Norwogonin (NWG), a natural plant flavone, has shown potential anti-inflammatory and antioxidative effects. However, whether it could ameliorate sepsis-related ALI and the potential mechanism remains unknown. PURPOSE:This study aims to investigate the effects and underlying mechanisms of NWG in treating sepsis-related ALI. METHODS:Male Wistar rats (200-220 g) were used to establish sepsis-related ALI model via intraperitoneal injection of lipopolysaccharide (LPS). Vital signs and arterial blood gas analysis, HE and immunohistochemistry staining, dynamic visualization of the microcirculatory system to observe FITC-dextran leakage and leukocyte adhesion, ELISA assay of inflammatory cytokines, Evans Blue extravasation, measurement of total protein content in bronchoalveolar lavage fluid, determination of the Wet/Dry weight ratio, Western blot and RT-qPCR analysis were used to evaluate NWG's effects and the potential mechanism. Additionally, we employed network pharmacology and molecular docking to identify and evaluate the interaction between NWG and the key targets of ALI. Surface plasmon resonance and enzyme activity assay were utilized to confirm the direct interaction between NWG and the potential targets. RESULTS:NWG administration improved the vital signs of LPS-stimulated rats. Exposure to LPS led to deteriorated arterial blood gas analysis, prominent lung morphology destruction, neutrophil and M1 macrophage infiltration, leukocyte adhesion, FITC-dextran leakage, elevated secretion of inflammatory cytokines, and aggravated lung edema. NWG intervention effectively mitigated these changes. Furthermore, NWG suppressed NF-κB/NLRP3 signaling and up-regulated endothelial junction proteins. Network pharmacology analysis and molecular docking identified five top key targets: MMP-9, AKT1, COX-2, Src and JAK-2. Western blot and RT-qPCR results confirmed that NWG inhibited the Src/AKT1/NF-κB signaling pathway, and down-regulated the levels of inflammatory factors. Surface plasmon resonance revealed the direct binding between NWG and AKT1, COX-2 and Src, rather than MMP-9. Enzyme activity assay demonstrated that NWG inhibited the activity of AKT1, COX-2 and Src. CONCLUSION:NWG alleviated inflammation, restored pulmonary microvascular barrier function and improved LPS-induced ALI. These effects were mediated by inhibiting the Src/AKT1/NF-κB signaling pathway through direct targeting of Src, AKT1 and COX-2. Our study provided novel scientific evidence supporting the use of NWG in the treatment of ALI caused by sepsis.

ETHNOPHARMACOLOGICAL RELEVANCE:Jianpi Qingre Chubi prescription primarily consists of a compound formula, also known as Huangqin Qingre Chubi Capsules (HQC), which strengthens the spleen and resolves dampness, clear heat, and collaterals. Long-term clinical use has shown that HQC improves joint swelling and pain in patients with osteoarthritis. Mechanistically, we demonstrated that HQC inhibits inflammatory responses, extracellular matrix degradation, and delays chondrocyte senescence. AIM:To determine the bioactivity and mechanism of action of Jianpi Qingre Tongluo prescription (HQC) on osteoarthritis (OA). MATERIALS AND METHODS:First, the chondroprotective effects of HQC were assessed using histopathology, immunohistochemical staining and protein blotting in an OA rat model. Additionally, we identified key targets for crucial targets of HQC in OA using the Network Pharmacology and Gene Expression Omnibus (GEO) dataset (GSE98918 and GSE152805). In vitro conditions, IL-1β-treated chondrocytes served to study the impact of HQC on OA development and the senescence-associated secretory phenotype (SASP). This was evaluated using a series of approaches, such as flow cytometry assays, and immunofluorescence staining, and then verified by rescue experiments. RESULTS:Therapy with HQC attenuated the severity of osteoarthritis (demonstrated by histopathology, OARSI grading scores, and Mankin scores) and SASP factors (as indicated by IL-1β, IL-6, IL-4, IL-37, MMP13, ADAMTS5, COL2A1, and ACAN levels, and apoptotic cell death). HQC might treat osteoarthritis via four important targets (STAG1, TP53, P21, and P16), with the p53 signalling pathway representing one of the main pathways. The HQC acts primarily on chondrocyte clusters. In vitro experiments indicated that STAG1 overexpression accelerates chondrocyte apoptosis, promotes SASP factor expression and extracellular matrix (ECM) degradation, and facilitates OA progression. HQC-containing serum suppressed the expression of the STAG1/TP53/P21 pathway, regulated SASP factors, and restored ECM balance. CONCLUSION:Jianpi Qingre Tongluo prescription modulated SASP factors by regulating the STAG1/TP53/P21 signal transduction axis and decelerating cartilage senescence and degradation in patients with OA. Jianpi Qingre Tongluo may be an effective drug candidate.

Background:Xinfeng Capsule (XFC) is a traditional Chinese medicine compound preparation that has been clinically used to treat rheumatoid arthritis (RA) for more than 20 years. It has demonstrated clear therapeutic effects, including improving pulmonary function and reducing lung injury in patients with RA. However, the precise mechanism underlying its protective effect against lung injury remains unclear. This study aims to explore the potential mechanisms of XFC in the treatment of lung injury. Methods:Liquid chromatography-mass spectrometry (LC-MS) analysis was conducted to determine the chemical composition of XFC. Proteomic and bioinformatic analyses of differentially expressed proteins (DEPs) in rat lung tissue were performed using tandem mass tag labeling. A rat adjuvant arthritis (AA) model was established using Freund's complete adjuvant to observe pathological changes in synovial and lung tissues, as well as alterations in lung function. In addition, a cell model was constructed by inducing lung fibroblasts with transforming growth factor-β1 (TGF-β1) to investigate the effects of XFC-containing serum on oxidative stress and pulmonary fibrosis through the peroxisome proliferator-activated receptor gamma (PPARγ)/3-hydroxy-3-methylglutaryl-CoA synthase 2 (HMGCS2) pathway. Results:LC-MS analysis identified a total of 867 compounds in XFC, of which 25 unique compounds were closely associated with pulmonary fibrosis and lung injury. Proteomic analysis suggested that XFC may regulate PPAR signaling pathway-related proteins and alleviate lung injury in AA rats. Animal experiments showed that XFC significantly inhibited immune inflammation, synovial hyperplasia, and oxidative stress in AA rats, while reducing lung injury and improving lung function. Furthermore, XFC-containing serum suppressed TGF-β1-induced proliferation of lung fibroblasts, promoted PPARγ expression, and significantly decreased the levels of interleukin-6, tumor necrosis factor-α, reactive oxygen species, nicotinamide adenine dinucleotide phosphate oxidase 4, HMGCS2, collagen type I α 1, collagen type III α 1, and α-smooth muscle actin (P < 0.01). In addition, XFC partially reversed the effects of the PPARγ antagonist GW9662, activated the PPARγ signaling pathway, inhibited oxidative stress and inflammatory responses, and exerted anti-fibrotic effects similar to those of the PPARγ agonist rosiglitazone. Conclusion:XFC inhibits inflammation and oxidative stress by regulating the PPARγ/HMGCS2 pathway, thereby attenuating fibrosis and alleviating lung injury.

Although doxorubicin (DOX) is an efficient chemotherapeutic drug for human tumors, severe cardiotoxicity restricts its clinical use. Oridonin (Ori), a bioactive component isolated from Isodon rubescens (Hemsl.) H. Hara, possesses potent anti-inflammatory and anticancer potentials. Therefore, our study aimed to evaluate the protective effects of Ori against DOX-induced cardiotoxicity. DIC models were established in vivo and in vitro. The action targets and pharmaceutical mechanism of Ori against DIC were comprehensively examined by network pharmacology, RNA-sequencing, and experimental validation. Ori relieved Dox-induced cell apoptosis in vitro and in vivo. A total of 7084 DEGs, 196 Ori, and 8172 DIC targets were screened by transcriptomics and network pharmacology, respectively. The three sets contained 11 intersection genes, including Ccl2, Myc, Mmp3, Egfr, p38 MAPK (MAPK14), Esr1, Tnf, Jun, Cdk1, Alb, and Ccnd1. The experimental results showed that Ori significantly decreased MMP-3 activity and the expression of p38 MAPK, thereby attenuating myocardial apoptosis and inflammatory infiltration. This study suggests that Ori is a potential therapeutic agent for DOX-induced cardiotoxicity that exerts its effects by inhibiting the p38 MAPK/MMP-3 signaling pathway.

BackgroundRheumatoid arthritis (RA) is often accompanied by abnormal changes in inflammatory responses and coagulation-fibrinolysis indicators. Jianpi Huashi Tongluo Prescription - Xinfeng Capsule (XFC), a traditional Chinese medicine formulation comprising multiple herbal ingredients, is widely used clinically for the treatment of RA. It exhibits dual anti-inflammatory and anticoagulant effects. However, the specific mechanisms underlying its actions remain to be further investigated.ObjectiveThis study aims to elucidate the anti-inflammatory and anticoagulant mechanisms of XFC in the treatment of RA.MethodsA multidimensional methodological framework was employed. Firstly, through retrospective clinical data mining, combined with the Apriori algorithm and random walk models, an in-depth analysis was conducted to explore the potential associations between XFC treatment and improvements in clinical inflammatory and coagulation markers among RA patients. Secondly, an adjuvant-induced arthritis rat model was established to directly observe the anti-inflammatory and anticoagulant effects of XFC in vivo. Furthermore, bioinformatics and network pharmacology techniques were applied to decipher the major active components and their targets of XFC. Lastly, a co-culture system of RA patient-derived peripheral blood mononuclear cells (RA-PBMCs) and vascular endothelial cells (VECs) was established to mimic the in vivo microenvironment, and the anti-inflammatory and anticoagulant mechanisms of XFC were validated in vitro.ResultsData mining analysis revealed abnormally elevated levels of inflammatory and coagulation markers such as fibrinogen (FBG), erythrocyte sedimentation rate (ESR), high-sensitivity C-reactive protein (Hs-CRP), and rheumatoid factor (RF) in RA patients (p < 0.001), and emphasized the close correlation between XFC treatment and the improvement of these markers including Hs-CRP, ESR, and RF (confidence >60% and lift >1). Animal experimental data indicated that XFC effectively reduced the levels of inflammatory and coagulant markers (IL-6, D-D, FBG, PAF, VEGF, and TF) in adjuvant-induced arthritis (AA) rats while enhancing the expression of anti-inflammatory factors (IL-10) (p < 0.05). Furthermore, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) results suggested that the pharmacodynamic mechanism of XFC may be closely related to the regulation of the PI3K/AKT signaling pathway. Additionally, network pharmacology and molecular docking results show that the main active components of XFC, namely, calycosin-7-O-beta-D-glucoside, calycosin, and formononetin, exhibit excellent docking with the core targets HIF1A, PTGS2, and MMP9. In vitro co-culture model showed that XFC inhibited RA-related inflammatory responses and hypercoagulable states by suppressing the activation of the PI3K/AKT signaling pathway.ConclusionThis study demonstrates that XFC exerts its dual anti-inflammatory and anticoagulant effects, at least in part, by inhibiting the activation of the PI3K/AKT signaling pathway, providing potential insights into targeted therapy for RA.

ETHNOPHARMACOLOGICAL RELEVANCE:YangXue QingNao Wan (YXQN) is a compound Chinese medicine comprising of 11 traditional Chinese medicinal herbs, including Angelica sinensis, Ligusticumstriatum, and Paeonia lactiflora, etc. Previous studies in our laboratory have demonstrated that YXQN improved cerebral microcirculation in hypertensive rats. However, its efficacy and underlying mechanisms in treating vascular dementia (VaD) remain unclear. AIM OF THE STUDY:This study aims to investigate the therapeutic effects and underlying mechanisms of YXQN on VaD using a rat model of bilateral common carotid artery occlusion (2-VO). MATERIALS AND METHODS:Male Wistar rats (10-12 weeks old) were randomly divided into seven groups: Sham, YXQN, 2-VO model, 2-VO with low-, medium-, or high-dose YXQN, and 2-VO with donepezil as a positive control. The 2-VO model was established by bilateral common carotid artery ligation, followed by 36 consecutive days of oral YXQN administration. Cognitive function was assessed using the Morris water maze and Y-maze tests, while hippocampal neuronal damage was evaluated by Nissl staining. Western blot analysis was performed to examine the types of neuronal cell death in the hippocampus. Transcriptome sequencing was conducted to explore potential molecular targets and mechanisms. In addition, in vitro experiments using HT22 cells were carried out to further validate the effects and mechanisms of YXQN. RESULTS:Our results showed that YXQN alleviated learning and memory impairments, as well as hippocampal neuronal structural damage in 2-VO rats. Western blot analysis indicated that ferroptosis occurred in the hippocampal neurons of 2-VO rats, characterized by a significant downregulation of GPX4 and NRF2, upregulation of TF, TFR and ACSL4, increased levels of Fe2+, MDA, 4-HNE and GSSG, decreased levels of GSH, and reduced expression of mitochondrial functional proteins ATP5A and ATP5D. All of these changes were reversed by YXQN treatment. The transmission electron microscopy (TEM) analysis revealed typical ferroptotic mitochondrial alterations in the hippocampal neurons of 2-VO group, which was largely improved by YXQN. Transcriptomic analysis revealed that the ferroptosis-related gene Dpp4 was upregulated in the 2-VO group and downregulated following YXQN administration. In vitro experiments confirmed that YXQN inhibited Erastin-induced ferroptosis in HT22 cells. Furthermore, Dpp4 knockdown significantly attenuated Erastin-induced downregulation of GPX4, while Dpp4 overexpression abolished YXQN-mediated upregulation of GPX4 expression in HT22 cells. Molecular docking analysis showed that Ursolic Acid (UA), an active compound in YXQN, strongly binds to Dpp4, and UA also inhibited Erastin-induced Dpp4 expression in HT22 cells. CONCLUSION:This study demonstrated that YXQN ameliorated cognitive dysfunction and hippocampal neuronal degeneration in 2-VO rats through inhibition of ferroptosis, with UA identified as a critical component mediating the inhibition of Dpp4. This study provided scientific evidence for the application of YXQN in VaD prevention and treatment.

Objectives Circular RNAs (circRNAs) are known to be associated with cardiovascular diseases. At present, an ideal biomarker for the early diagnosis of coronary heart disease (CHD) is still lacking.Methods We screened differentially expressed circRNAs (DEcircRNAs) in the peripheral blood monocytes (PBMCs) of patients with CHD, using the microarray technology in comparing the transcriptome. We identified upregulated and downregulated circRNAs. At the same time, we collected the patient clinical medical records and the PBMCs, the above results were analyzed and validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), using 374 patients.Results We identified 183 upregulated and 41 downregulated circRNAs. Among these DEcircRNAs, hsa_circ_0000745/hsa_circRNA_101996 was significantly upregulated in a cohort of 297 patients with CHD and 77 non-CHD controls. Among patients with CHD, hsa_circ_0000745/hsa_circRNA_101996 was significantly upregulated in the unstable angina pectoris (UAP) and acute myocardial infarction (AMI) subgroups compared to the stable angina pectoris (SAP) subgroup. By dividing hsa_circ_0000745/hsa_circRNA_101996 expression into quartiles, we observed that the highest hsa_circ_0000745/hsa_circRNA_101996 expression quartile was a risk factor for CHD compared to the lowest quartile (odds ratio [OR]: 2.709; 95 % confidence interval [CI]: 1.126-6.519, p=0.026), after adjusting for the traditional risk factors (age, sex, body mass index [BMI], smoking, alcohol, C-reactive protein [CRP], small and dense low-density lipoprotein [sdLDL] and lipoprotein-associated phospholipase A2 [LP-PLA2]).Conclusions These data suggest that upregulated hsa_circ_0000745/hsa_circRNA_101996 in PBMCs is a risk factor for CHD and could be used as a biomarker of CHD.

ETHNOPHARMACOLOGICAL RELEVANCE:Lizhong decoction (LZD), a Traditional Chinese Medicine formula, is widely utilized to treat gastrointestinal diseases, including ulcerative colitis in China for thousands of years. AIM OF THE STUDY:To investigate whether the protective effect of LZD on ulcerative colitis is dependent on gut microbiota and T-cell immune homeostasis. MATERIAL AND METHODS:The preventive effects of LZD on dextran sodium sulfate (DSS)-induced colitis mice were evaluated through the measurement of body weight, disease activity index, colon length and hematoxylin-eosin staining. Flow cytometry was used to detect the ratio of Th17/Treg cells. Pseudo sterile mice and fecal transplantation experiments were used to investigate whether the preventive effect of LZD was dependent on the gut microbiota. The alterations of gut microbiota were identified by the 16S rDNA sequencing. The content of intestinal short-chain fatty acids (SCFAs) was detected by LC-MS/MS analysis. The downstream signal pathways of SCFAs were detected by the immunoblotting. RESULTS:LZD administration significantly alleviated weight loss and intestinal injury in DSS-induced colitis mice. LZD administration also promotes the balance of Th17/Treg cells. Moreover, LZD administration relies on gut microbiota to alleviate ulcerative colitis and regulate Th17/Treg cell balance. LZD administration significantly improves gut microbial composition in colitis mice, elevating the abundance of SCFAs producing bacterium such as lachnospiraceae_nk4a136_group and Akkermansia. LZD treatment further increases the abundance of SCFAs and promotes activation of free fatty acid activated receptor 2 (FFAR2). CONCLUSION:LZD administration promotes Th17/Treg cell balance in a gut microbiota-SCFAs dependent manner, which in turn ameliorates ulcerative colitis.