Objective To investigate the effects of TLR7/8 agonist Resiquimod (R848) on the generation and immunosuppressive function of γδ Treg induced by TGF-β1, and explore the underlying molecular mechanisms. Methods γδ T cells were expanded with anti-human γδ TCR antibodies in the presence of different concentrations of TGF-β1, and the expression of transcription factor forkhead box protein P3 (FoxP3) in γδ T cells was subsequently analyzed by flow cytometry. Then Vδ1 and Vδ2 γδ T cells were amplified by anti-human Vδ1 TCR anti- body and anti-human γδ TCR antibody respectively, and R848 was added to the induction system to examine its effect on the proportion of FoxP3⁺γδ Treg. Induced Vδ1 and Vδ2 γδ T cells were isolated by magnetic-activated cell sorting, and the effect of R848 on their suppressive function against CD4⁺ T cell proliferation was assessed by a CFSE dilution assay. Flow cytometry was used to detect the expression of perforin and granzyme B. Western blot was performed to measure the expression level of granzyme K. ELISA was used to determine the concentration of TGF-β1 in the supernatant, and RNA sequencing was performed to identify differentially expressed genes after R848 treatment with validation using Western blot. Results 3 ng/mL TGF-β1 significantly increased the expression of FoxP3 in γδ T cells. R848 had no significant effect on the generation of TGF-β1-induced FoxP3⁺ Vδ1 γδ Treg, but significantly reduced the proportion of FoxP3⁺ Vδ2 γδ Treg. R848 treatment reversed the suppressive effect of Vδ2 γδ Treg on CD4⁺ T cell proliferation, whereas no such effect was observed in Vδ1 γδ Treg. R848 partially reversed the suppressive function of Vδ2 γδ Treg, promoted granzyme B expression, and significantly decreased their secretion of TGF-β1.RNA-seq analysis and Western blot validation demonstrated that R848 reversed the suppressive function of Vδ2 γδ Treg by downregulating the expression of SOCS1 and CISH, two key negative regulators of the JAK/STAT signaling pathway and inhibiting STAT3 phosphorylation. Conclusions The TLR7/8 agonist R848 reverses the generation and suppressive function of TGF-β1-induced Vδ2 γδ Treg. These findings provide experimental support for the development of a combinational immunotherapeutic strategy involving γδ T cells and R848.

Objective To investigate the effects of the T204A variant in triggering receptor expressed on myeloid cells 2(TREM2) on microglial phagocytic function and the activity of the TREM2-DAP12-SYK signaling pathway. Methods Wild-type and mutant constructs were expressed in 293T and HMC3 microglia. Protein expression and localisation were assessed by immunofluorescence and Western blot. Aβ clearance-related capacity was evaluated by measuring residual Aβ levels in the supernatant using ELISA. TREM2-DAP12 interaction was analysed by Co-IP, and SYK/AKT phosphorylation was examined by Western blot. Results This study identified three SNPs in the TREM2 coding region, two of which were located in isoform2 and one in isoform1. None of the three SNPs affected TREM2 expression or localisation. Only T204A significantly reduced residual Aβ levels in the supernatant(P＜0.05), indicating enhanced Aβ clearance-related capacity. Mechanistically, T204A increased TREM2-DAP12 interaction and was associated with elevated SYK phosphorylation. Conclusions TREM2 T204A is a gain-of-function variant that enhances TREM2-DAP12 complex formation and is associated with increased SYK signaling and Aβ clearance in microglia.

Objective To investigate the regulatory mechanisms of the cyclic adenosine monophosphate (cAMP) signaling pathway in the pathogenesis of silicosis by screening and identifying novel proteins that interact with cAMP in mouse alveolar macrophage cells (MH-S) stimulated with silica. Methods MH-S cells were divided into a control group and an experimental group. Intracellular cAMP levels were measured using enzyme-linked immunosorbent assay (ELISA). Pull-down assays were performed with cAMP-conjugated agarose beads to capture cAMP-binding proteins. These proteins were separated by SDS-PAGE and visualized by silver staining. Liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteomics was used to screen for potential cAMP-binding proteins. AlphaFold was used to predict the structures and assess the binding capacities of selected candidate proteins. Results Compared with the control group, the intracellular cAMP level in the experimental group was significantly decreased (P＜0.01) and the profile of cAMP-binding proteins was altered. A total of 5 upregulated and 413 down- regulated proteins were identified. AlphaFold predictions indicated that the upregulated proteins might be potential cAMP-binding proteins with high confidence. Conclusions Silica stimulation can alter the intracellular cAMP levels and profile of cAMP-binding proteins in alveolar macrophages. These novel cAMP-binding proteins may participate in the occurrence and progression of silicosis by increasing expression levels or enhancing binding affinity to cAMP, providing a new avenue for the discovery of potential therapeutic targets for silicosis.

Objective To investigate the regulatory role of histone deacetylase 4(HDAC4) overexpression in the proliferation and invasion of human colorectal cancer (CRC) cells. Methods The expression level of HDAC4 in CRC and its correlation with patient prognosis were analyzed using TCGA, GTEx, and HPA databases. Lentiviral transfection was used to construct HDAC4 overexpressing CRC cell lines (SW480 and HCT116), with transfection efficiency verified by RT-qPCR and Western blot. The effects of HDAC4 overexpression on CRC cell proliferation and invasion were assessed using CCK-8, EdU, colony formation, and Transwell assays. Results Database analysis revealed that HDAC4 was significantly downregulated in human CRC tissues and closely associated with poor patient prognosis. Immunohistochemical validation using HPA clinical samples further confirmed that HDAC4 expression was markedly lower in CRC tissues compared to normal colorectal tissues. RT-qPCR and Western blot confirmed the successful establishment of stable HDAC4 overexpressing SW480 and HCT116 cells. HDAC4 overexpression reduced CRC cell viability (P＜0.001), decreased the number of EdU-positive cells (P＜0.01), and significantly inhibited colony formation (P＜0.001). Additionally, HDAC4 overexpression downregulated the expression of proliferation-related genes PCNA(P＜0.01) and MK167(P＜0.01) and significantly impaired cell invasion ability. Conclusions HDAC4 overexpression suppresses the proliferation and invasion of human CRC cells, providing new theoretical and experimental support for precision therapy in colorectal cancer.

Objective To investigate the effects of Pifithrin-μ on the proliferation of TSC2 mutant hepatocellular carcinoma cells. Methods The effect of Pifithrin-μ on cell proliferation was evaluated using CCK8 assay in multiple lines of Tsc2 mutant mouse embryonic fibroblasts (MEFs) and human hepatocellular carcinoma cells. Lentiviral infection was used to construct hepatocellular carcinoma cells with knockdown or overexpression of TSC2. The protein expression of TSC2 and the downstream effectors of the mTOR pathway were detected by Western blot. Cellular sensitivity to Pifithrin-μ was detected by CCK8. The nude mouse subcutaneous transplantation tumor model was established for evaluation of drug efficacy in vivo. RNA sequencing was used for differential gene and pathway enrichment analysis, which was verified by Western blot and CCK8. Results Pifithrin-μ selectively inhibited the proliferation of Tsc2 mutated MEFs and hepatocellular carcinoma cells in a dose-dependent manner(P＜0.05). Hepatocellular carcinoma cells with TSC2 knockdown showed increased sensitivity to Pifithrin-μ, whereas those with TSC2 overexpression were resistant (P＜0.05). Pifithrin-μ significantly inhibited the growth of Tsc2 mutant transplantation tumors in nude mice (P＜0.05). Pifithrin-μ downregulated the expression of Smad7 protein in Tsc2 mutant cells, resulting in elevated levels of p-Smad2/3 and over-activation of the TGF-β/Smad signaling pathway (P＜0.05). The combination of each of the six cell death inhibitors with Pifithrin-μ revealed that reactive oxygen species inhibitor and TGF-β/Smad pathway inhibitor were able to reverse Pifithrin-μ-induced cell death in TSC2 mutant cells (P＜0.05). Conclusions Pifithrin-μ selectively inhibits the proliferation of TSC2 mutant hepatocellular carcinoma cells. This effect is associated with the downregulation of Smad7 and the disruption of the TGF-β/Smad signaling pathway balance.

Objective Cancer-associated fibroblasts(CAFs) represent the predominant stromal cell population in the tumor microenvironment of pancreatic ductal adenocarcinoma (PDAC) and play critical roles in tumor progression and the development of therapeutic resistance. CAFs not only participate in stromal remodeling through abnormal proliferation and migration but also regulate the tumor microenvironment by secreting various metabolites that support tumor cell growth. Carboxyamidotriazole orotate(CTO) is a Complex Ⅰ inhibitor with antitumor activity; however, its effects on the biological functions and metabolic secretory profile of CAFs within the tumor microenvironment remain unclear. The present study aims to investigate the regulatory effects of CTO on CAF biological functions and metabolic secretion in pancreatic cancer and to explore its potential role in remodeling the tumor microenvironment. Methods SRB assays and Western blot analysis were performed to evaluate the effects of CTO on CAF proliferation and activation status. Transcriptome sequencing was used to analyze the alterations in CAF-related biological pathways following CTO treatment. In addition, metabolomic analysis of cell culture supernatants was conducted to characterize changes in the secreted metabolite profile of CAFs, thereby assessing the impact of CTO on metabolic support within the tumor microenvironment. Results CTO treatment significantly inhibited CAF proliferation and activation. Transcriptomic analysis revealed that CTO regulated multiple signaling pathways associated with stromal remodeling and tumor-promoting functions in CAFs, thereby attenuating their pro-tumor activity. Metabolomic analysis further demonstrated that CTO altered the composition of metabolites secreted by CAFs and affected the supply of key metabolites in the tumor microenvironment, consequently reducing the metabolic support provided by CAFs for tumor cell survival and growth. Conclusions CTO can remodel the pancreatic cancer tumor microenvironment by inhibiting CAF proliferation and regulating their metabolic secretory profile, thereby influencing tumor progression. These findings provide new experimental evidence supporting strategies that target stromal cells to modulate the tumor microenvironment.

Objective To establish an animal model of chronic pancreatitis-induced osteoporosis and to investigate the effects of chronic pancreatitis (CP) on bone loss in mice and the underlying mechanism. Methods Forty Kunming (KM) mice aged 6-8 weeks were divided into control group (n=20) and CP group (n=20). The CP mouse model was established by intraperitoneal injection of L-arginine, while the control group received an equal volume of normal saline. Samples were collected at 8 and 12 weeks after modeling. Pathological changes in the pancreas were observed by HE staining and Masson staining. Bone quality and bone morphology were assessed by micro-CT and HE staining. The expression of bone formation markers OPN and Runx2 proteins was detected by immunohisto- chemical staining. Levels of fecal elastase-1, serum vitamin D, Ca²⁺, and osteocalcin were measured by ELISA. Results At 8 weeks after modeling, compared with the control group, the CP group showed atrophy and necrosis of the pancreatic parenchyma, accompanied by extensive inflammatory cell infiltration and increased fibrosis; decreased trabecular bone density in the femur and tibia, significant destruction of bone microstructure, and reduced overall bone mass (P＜0.05); and significantly decreased expression of bone formation markers OPN and Runx2 proteins in bone tissue (P＜0.05). The CP group also exhibited significantly lower levels of fecal elastase-1, serum vitamin D, Ca²⁺, and osteocalcin (P＜0.05). These changes became more significant with the extension of modeling duration to 12 weeks. Conclusions Intraperitoneal injection of L-arginine successfully establishes a mouse model of chronic pancreatitis-induced osteoporosis. The pathological mechanism is associated with impaired vitamin D absorption leading to reduced bone formation.

Objective To investigate the therapeutic effect of the natural flavonoid taxifolin in experimental pulmonary arterial hypertension (PAH), and to further explore its potential molecular mechanisms underlying the amelioration of PAH. Methods This study first employed network pharmacology to predict the potential targets of taxifolin in PAH. After establishing a PAH model with monocrotaline (MCT), rats were administered with taxifolin or vehicle. Hemodynamic parameters, right ventricular hypertrophy, and pulmonary vascular remodeling were evaluated. Transcriptomic sequencing and analysis were performed to investigate the underlying molecular mechanisms, with results validated through immunofluorescence staining. Results Network pharmacology indicated that taxifolin was closely associated with biological processes related to leukocyte chemotaxis and vascular regulation. In MCT-induced PAH rats, taxifolin significantly reduced right ventricular systolic pressure (P＜0.001), attenuated right ventricular hypertrophy (P＜0.05), and inhibited pulmonary arteriolar remodeling (the number of muscularized vessels was significantly reduced, P＜0.000 1, and the medial wall thickness of small vessels was significantly decreased, P＜0.000 1). Transcriptomic analysis revealed that taxifolin intervention selectively reversed the aberrant expression of PAH-related chemokine-binding genes and regulated the expression of atypical chemokine receptors such as Ackr1 and Ackr2, suggesting that it may exert pulmonary vascular protection through regulating chemokine signaling. Conclusions Taxifolin ameliorates experimental PAH, showing disease-modifying effects in attenuating pulmonary vascular remodeling, potentially through the modulation of leukocyte chemotaxis. These findings suggest the therapeutic potential of taxifolin and highlight the chemotaxis pathway as a tractable target.

Objective To clarify the regulatory effect of DNA released by apoptotic multiple myeloma cells on the antiviral innate immunity of macrophages, and to elucidate its molecular mechanism, to provide experimental evidence for understanding the immunological mechanism underlying the high susceptibility to viral infections in patients with multiple myeloma. Methods Human RPMI-8226 cells were induced to undergo apoptosis by ultraviolet irradiation. The apoptotic conditioned medium (Apo-CM) was collected to treat THP-1 macrophages, followed by VSV-GFP virus infection. Viral replication, as well as the expression levels of IFN-β, IL-6 and VSV-G were detected by fluorescence microscopy and quantitative real-time PCR (qPCR). DNase Ⅰ and RNase A were used to treat Apo-CM to identify the key effector molecules. Specific inhibitors targeting cGAS, STING and TLR9 were applied to block the corresponding pathways for further exploration of the immune regulatory mechanism. Results Preconditioning with Apo-CM markedly promoted viral replication and suppressed IFN-β secretion in macrophages, and such immunosuppressive effects were enhanced in a time-dependent manner (P＜0.05). Degradation of DNA by DNase Ⅰ significantly reversed the immunosuppressive effect of Apo-CM(P＜0.01), while RNase A treatment exerted no obvious influence. ConclusionsDNA derived from apoptotic myeloma cells inhibits the type Ⅰ interferon response and impairs antiviral innate immunity in macrophages through activating the TLR9 and cGAS-STING pathways, thereby facilitating viral replication.

Objective To investigate the effect of esketamine (S-KET) on pain in knee osteoarthritis (KOA) rats by modulating the TLR4/TNF-α/MMP-9 signaling pathway. Methods A KOA rat model was established by intra-articular injection of papain. Successfully modeled rats were assigned into KOA group, L-S-KET, M-S-KET, H-S- KET groups (intraperitoneal injection of 10, 20, and 40 mg/kg S-KET), and H-S-KET+LPS group (intraperitoneal injection of 40 mg/kg S-KET+intra-articular injection of 0.25 mg/kg TLR4 activator LPS) according to the random number table method, with 10 rats in each group. Another 10 normal rats served as control group. The control group and KOA group were given equal amounts of physiological saline once a day for 28 consecutive days. The thermal radiation method and pain threshold measurement instrument were used to detect the pain threshold of rats. HE staining was used to observe pathological changes in rat cartilage tissue, and the pathological scoring was performed. TUNEL staining was used to observe the apoptosis of chondrocytes in cartilage tissue. ELISA was performed to detect IL-6, TNF-α, and IL-1β in serum. Western blot was performed to detect changes in the expression of TLR4/TNF-α/MMP-9 signaling pathway proteins in joint tissues. Results Compared with the control group, the cartilage tissue of rats in KOA group lost normal structure and the Mankin score increased (P＜0.05). Compared with KOA group, the L-S-KET group, M-S-KET group, and H-S-KET group showed reduced cartilage damage, relatively uniform cell arrangement, and decreased Mankin score(P＜0.05). Compared with the H-S-KET group, the H-S-KET+LPS group showed cracks in the cartilage tissue of rats, uneven cell arrangement, and an increase in Mankin score (P＜0.05). Compared with the control group, the KOA group had lower thermal pain threshold and mechanical pain threshold, higher chondrocyte apoptosis rate, IL-6, TNF-α, IL-1β levels, and TLR4, TNF-α, MMP-9 protein expression (P＜0.05). Compared with the KOA group, the L-S-KET group, M-S-KET group, and H-S-KET group had higher thermal pain threshold and mechanical pain threshold, lower chondrocyte apoptosis rate, IL-6, TNF-α, IL-1β levels, and TLR4, TNF-α, MMP-9 protein expression(P＜0.05). Compared with the H-S-KET group, the H-S-KET+LPS group had lower thermal pain threshold and mechanical pain threshold, higher chondrocyte apoptosis rate, IL-6, TNF-α, IL-1β levels, and TLR4, TNF-α, MMP-9 protein expression(P＜0.05). Conclusions S-KET can reduce inflammatory factors and alleviate KOA pain in rats. This may be achieved by modulating the TLR4/TNF-α/MMP-9 signaling pathway.

Objective To explore the impact of miR-130a-3p on the sensitivity of human cervical cancer cell line Caski to chemoradiotherapy. Methods The cells were divided into four groups: the control group (Caski cell line), cisplatin-resistant group (Caski/DDP cells), miR-130a-3p overexpression group (Caski+miR-130a-3p cells), and cisplatin-resistant miR-130a-3p overexpression group (Caski/DDP+miR-130a-3p cells). After treatment with various concentrations of cisplatin and radiotherapy, cell proliferation was assessed using the CCK-8 assay, cell apoptosis was measured by flow cytometry, and the expression of ZEB1 protein was detected by Western blot. Results After overexpression of miR-130a-3p, the IC₅₀ of cisplatin in Caski cells decreased significantly (9.447 μg/mL vs. 6.429 μg/mL, P＜0.05), and the apoptosis rate increased, although the difference was not statistically significant (mean apoptosis rate 29.3% vs. 30.6%, P＞0.05), and the expression of ZEB1 was significantly downregulated (P＜0.01). In Caski/DDP cell strain, the IC₅₀ decreased more markedly after overexpression of miR-130a-3p (IC₅₀: 58.38 μg/mL vs. 18.38 μg/mL, P＜0.000 1); the rate of cell apoptosis increased significantly (mean apoptosis rate: 15.9% vs. 29.8%, P＜0.001), and the expression of ZEB1 protein was downregulated (P＜0.01). Conclusions Overexpression of miR-130a-3p may increase the chemoradiotherapy sensitivity of cervical cancer cells by downregulating ZEB1 expression.

Objective To explore the role of CBP/p300-interacting transactivator 2 with Glu/Asp rich carboxy-terminal domain 2(CITED2) in patients with coronary slow blood flow (CSF) and its regulatory mechanism on endothelial cell hypoxia-reoxygenation (H/R) injury. Methods Peripheral blood samples were collected from CSF patients and healthy controls. Serum TNF-α, IL-6 and ROS levels were detected by ELISA, and the expressions of CITED2, hypoxia-inducible factor-1 α(HIF-1α), Dynamin-related protein 1(DRP1), Sirtuin 1(SIRT1) and Mitofusin 2(MFN2) in peripheral blood mononuclear cells (PBMCs) were analyzed by RT-qPCR and Western blot. Human umbilical vein endothelial cells (HUVECs) were cultured in vitro to construct an H/R model, and the expression of CITED2 was regulated by transfection of siRNA and overexpression plasmid. The experiment was divided into control group, H/R group, H/R+OE-NC group, H/R+OE-CITED2 group, H/R+siNC group and H/R+siCITED2 group. The intracellular ROS level and mitochondrial membrane potential were detected by DHE and JC-1 staining, and the expression of related genes and proteins was detected by RT-qPCR and Western blot. Results Compared with the control group, the levels of TNF-α, IL-6 and ROS in CSF patients were significantly increased(P＜0.05), the expressions of HIF-1α and DRP1 in PBMCs were upregulated, and the expressions of CITED2, SIRT1 and MFN2 were down-regulated(P＜0.05). In HUVECs, the expression of HIF-1α and DRP1 increased significantly after H/R treatment, while the expression of CITED2, SIRT1 and MFN2 decreased. Overexpression of CITED2 significantly inhibited the expression of HIF-1α and DRP1, upregulate SIRT1 and MFN2, reduce ROS production and restore mitochondrial membrane potential. Knockdown of CITED2 aggravated the above damage effect. Conclusions CITED2 can reduce ROS production and restore mitochondrial membrane potential by regulating the expressions of HIF-1α, DRP1, SIRT1 and MFN2, thus improving the endothelial cell injury associated with CSF.

Objective To explore the efficacy of plan-do-check-act(PDCA)cycle management in improving the detection efficiency of the BRAF V600E mutation in fine needle aspiration cytology (FNAC) samples. Methods 275 FNAC samples received from September to December 2023 (before PDCA improvement) by the Department of Pathology of Xuanwu Hospital were collected as the control group, and 406 samples received from January to March 2024 (after PDCA improvement) were collected as the study group. The positive detection rate, assay repeat rate, and timely reporting rate of the two groups were compared to evaluate the effect of PDCA management. Results The positive detection rate increased from 45.45% before implementation to 55.17% after implementation. The assay repeat rate decreased from 6.18% (17/275) to 1.72% (7/406)(P＜0.05). The average turnaround time (TAT) for reporting results was reduced from (1.85±1.35)working days to (1.45±0.86)working days. Consequently, the timeliness reporting rate improved from 87.64% to 94.83%(P＜0.05). Conclusions Improving the whole process of molecular detection through PDCA management can effectively improve the accuracy and timeliness of BRAF V600E detection, which can help form a long-term mechanism to ensure the continuous improvement of molecular detection quality.

Objective To report and analyze the clinical characteristics of a child with Langer-Giedion syndrome (LGS) combined with Cornelia de Lange syndrome type 4 (CdLS4), and to review the literature in order to improve understanding of the clinical phenotypes and differential diagnosis of contiguous gene deletion syndromes. Methods A retrospective analysis was conducted on the clinical data of one patient with LGS co-occurring with CdLS4 who was admitted to the Department of Pediatrics, Peking Union Medical College Hospital in October 2024. Relevant literature was also reviewed. Results The patient was an 8-year-old boy whose main clinical features included characteristic facial appearance (thick eyebrows, mild synophrys, long eyelashes, prominent ears, broad nasal bridge, bulbous nasal tip, long philtrum, and thin upper lip), language developmental delay, intellectual disability, multiple osteochondromas, and skeletal abnormalities. Genetic testing revealed a heterozygous copy number deletion of approximately 2.2 Mb in chromosome 8q23.3-q24.12, involving the RAD21, EXT1, and TRPS1 genes, which was classified as pathogenic. He was diagnosed with Langer-Giedion syndrome combined with Cornelia de Lange syndrome type 4. Literature review showed that when deletions involve multiple genes in this region, patients may present overlapping phenotypes of both syndromes. In the early stages, reliance solely on clinical manifestations makes differentiation difficult, leading to a risk of misdiagnosis or delayed diagnosis. Conclusions LGS and CdLS4 share partially overlapping clinical phenotypes, and contiguous gene deletions can result in combined manifestations of multiple syndromes. Genetic testing enables definitive diagnosis and helps avoid misdiagnosis. This case highlights the importance of comprehensive genetic evaluation in children with developmental delay and multiple congenital anomalies, and the need for individualized long-term management plans under multidisciplinary collaboration.

Objective To explore the treatment strategy for patients with locally advanced rectal cancer harboring a POLE proofreading deficiency mutation. Methods The clinical data of a patient with locally advanced rectal cancer carrying the POLE p.V411L mutation was analyzed. Results The patient was a 41-year-old male with mismatch repair-proficient locally advanced rectal cancer. After initially receiving neoadjuvant concurrent chemoradiotherapy, laparoscopic exploration revealed that the tumor was unresectable. Subsequent genetic testing identified a POLE proofreading deficiency mutation with a tumor mutational burden of 426 mut/MB. The patient then underwent two years of treatment with envafolimab, achieving long-term complete clinical remission. Conclusions Colorectal cancer with POLE proofreading deficiency mutation is highly sensitive to immunotherapy. Genetic testing should be considered to screen for POLE proofreading deficiency mutations and other immunotherapy-sensitive variants in cases where rectal cancer invades surrounding tissues or organs, or when organ function preservation is a concern during surgery.

Pulmonary arterial hypertension(PAH) is characterized by pulmonary vascular remodeling, which is difficult to reverse with current therapies. Natural bioactive components provide a novel strategy for PAH intervention due to their multi-target advantages. This review systematically summarizes the mechanistic characteristics of natural bioactive components against PAH, encompassing multi-dimensional network effects such as improving vasomotor function, inhibiting smooth muscle cell proliferation, modulating inflammatory responses, and correcting metabolic reprogramming. Furthermore, it profoundly analyzes the evidence limitations in current research regarding direct molecular target validation, experimental models, structural endpoint evaluation, and druggability. Finally, we provide translational perspectives on strengthening target validation, upgrading pre-clinical evaluation systems, and optimizing delivery systems, aiming to provide a reference for promoting the substantive clinical translation of natural bioactive components in PAH.

Autoimmune hepatitis (AIH) is a chronic, inflammatory liver disease mediated by the immune system. Due to a breakdown in immune tolerance to liver autoantigens, multiple immune cells become activated and release abundant inflammatory factors. These autoantigens and inflammatory factors act upon cell surface receptors, activating multiple signaling pathways, including NF-κB, AMPK, PI3K/Akt, JAK/STAT, and MAPK, which induce or exacerbate immune-mediated liver injury. This occurs through the modulation of various processes in immune cells (such as dendritic cells, macrophages, and T cells), including energy metabolism, autophagy, apoptosis, and receptor expression.

Copper homeostasis imbalance and cuproptosis exacerbate pulmonary inflammation by promoting the release of cytokines through oxidative stress and mitochondrial damage; contribute to pulmonary hypertension associated with chronic obstructive pulmonary disease (COPD) by participating in vascular endothelial growth factor (VEGF)-mediated angiogenesis; worsen asthma symptoms by enhancing the activity of the thioredoxin-interacting protein (TXNIP) signaling pathway; induce pulmonary fibrosis by activating epithelial-mesenchymal transition (EMT) signaling; and accelerate the growth and metastasis of lung cancer by upregulating hypoxia inducible factor (HIF)-driven angiogenesis. Clinical translation of these mechanisms may provide novel insights and therapeutic strategies for the diagnosis and treatment of pulmonary diseases.

Sepsis, a life-threatening inflammatory syndrome, involves immune dysfunction, vascular homeostasis imbalance, and metabolic reprogramming. Abnormal lactate accumulation is a hallmark metabolic feature. Emerging evidence demonstrates that lactate-mediated protein lactylation contributes to sepsis progression by regulating immune cell activation, inflammatory factor expression, metabolic pathways, and cell death. Specifically, lactylation modulates the release of inflammatory factors and immune cell activity, mediates metabolic state remodeling, and influences vascular endothelial function and programmed cell death, thereby playing a critical regulatory role in sepsis pathophysiology. Although current studies link lactylation to sepsis-related inflammation, metabolic disturbances, and organ dysfunction, its integrated regulatory framework during sepsis progression remains incompletely defined. Furthermore, the theoretical basis for targeting lactylation therapeutically requires further clarification. This review systematically summarizes the regulatory functions and molecular mechanisms of lactylation in sepsis, focusing on inflammation and immune responses, metabolic reprogramming, vascular homeostasis, and programmed cell death. Our aim is to indicate subsequent mechanistic investigations and the development of targeted therapeutic interventions.

Small cell lung cancer (SCLC) is an aggressive pulmonary malignancy with a poor prognosis. Advances in PET/CT imaging and artificial intelligence have improved the accuracy of clinical staging and enabled more precise prediction of treatment responses. Additionally, circulating tumor DNA analysis and molecular subtyping have deepened our understanding of SCLC, laying a theoretical foundation for precision diagnosis and therapy. Therapeutically, immune checkpoint inhibitors have significantly improved outcomes for SCLC patients. Moreover, targeted therapies—such as DLL3 inhibitors, anti-angiogenic agents, PARP inhibitors, and AAK inhibitors—along with novel therapeutics, are opening new avenues and strategies for SCLC management. This review summarizes the major diagnostic and therapeutic advances in SCLC over the past five years, to provide reference and insights for both basic and clinical research.

Objective To investigate the current status of pain medicine education and career plan among medical students undergoing clinical training at a tertiary teaching hospital in Beijing through a questionnaire survey. Methods A self-designed questionnaire was distributed to and completed voluntarily by medical students during their rotation in the anesthesiology department. Results A total of 65 valid responses were collected (response rate: 92.86%), including 26 junior internship students and 39 senior internship students. Among them, 62 students (95.38%) expressed willingness to continue working as doctors after graduation, 2 students were interested in specializing in pain medicine, and 1 student planned to pursue a career in pain medicine abroad. Nine students (13.85%) reported having received pain medicine training during their clinical rotations, but 88.89% of them felt the training content was insufficient to meet clinical needs. Fifty students (76.92%) expressed a desire to participate in a pain medicine rotation, preferably lasting 1-2 weeks, to observe pain clinics, interventional treatments,and postoperative pain management. Conclusions Medical students lack sufficient knowledge in pain management, which is insufficient to meet the increasing demands of social healthcare services. In the future, incorporating mandatory/elective theoretical courses and clinical rotations in pain medicine could provide students with learning opportunities and foster greater awareness and emphasis on patient pain management in their professional practice.

Objective To explore the significance of surgical skills training in general practitioner education, the existing problems, and solutions. Methods A questionnaire survey was conducted among general practitioners undergoing and those who had completed standardized residency training at the Department of General Medicine, Peking Union Medical College Hospital from August 2014 to May 2022, as well as their instructors. The questionnaire included the respondents′ basic information, evaluation of the importance of surgical training, perceived current problems in surgical training, and potential solutions to these problems. The questionnaire was distributed and completed using the “Questionnaire Star” software. Results Surgical skills training was an essential part of general medical education. General practitioners identified general surgery and orthopedics as key departments requiring focused training. Additionally, specific surgical skills, such as wound dressing and suture removal, were crucial for general practitioners. However, the current surgical training model faced issues such as limited clinical practice and lack of practical opportunities. Conclusions Surgical skills training is a vital component of general medical education, contributing to the development of general practitioners′ surgical clinical practice capabilities. Nonetheless, the current methods of surgical skills training have many deficiencies. Understanding general practitioners′ perceptions of surgical training can help educational administrators design and implement targeted surgical skills training programs.

With changes in the medical model, restructuring of healthcare systems, and rapid development of digital technologies, clinical research is shifting from a paradigm centered on single projects and classical study designs toward a new paradigm emphasizing real-world settings, multisource data, interdisciplinary collaboration, and evidence translation. In contrast, current medical education still mainly focuses on disease knowledge, traditional epidemiology and biostatistics, and basic biomedical techniques, with insufficient training in research planning, systems thinking, teamwork, and translation. Internationally, platform-based, competency-oriented, and system-embedded training models have gradually emerged in clinical and translational science, learning health systems, implementation science, clinical informatics, precision medicine, and real-world research. In China, under the background of New Medical Education, training has begun to expand from traditional clinical research methods courses to comprehensive literacy, translational capacity, and interdisciplinary competence, but overall remains in a transitional stage from policy guidance and local exploration to systematic construction. In particular, the rapid emergence of generative artificial intelligence and AI agents in medical education, health care, and scientific research is accelerating changes in clinical research processes and organization, while also making the abilities to raise high-quality questions, generate original hypotheses, and maintain scientific judgment increasingly critical. This article reviews the current status of relevant education at home and abroad, analyzes major problems, and proposes suggestions for curriculum and training reform, to provide reference for the cultivation of clinical research talents in China.