ObjectiveThis study aims to elucidate the potential mechanism of Zuojinwan in improving liver fibrosis through hepatic tissue metabolomics analysis. MethodsTwenty-four mice were randomly allocated into normal group， model group ， positive drug group （silymarin， 100 mg·kg^-1）， and Zuojinwan group （Zuojinwan solution， 2.5 g·kg^-1）， with per group six mice. Liver fibrosis model was induced via intraperitoneal injection of olive oil solution with 10% carbon tetrachloride （CCl₄） （0.5 μL·g^-1， three times weekly for 8 weeks） in all groups except the normal group. During the final 4 weeks， the silymarin group received silymarin （100 mg·kg^-1） by gavage thrice weekly， while the Zuojinwan group was administered Zuojinwan solution （2.5 g·kg^-1） under the same regimen. After the last administration， the levels of liver fibrosis indicators and liver injury markers in serum were detected. The pathological morphological changes of the liver tissues were observed. The levels of liver fibrosis markers α-smooth muscle actin （α-SMA） and Collagen Ⅰ（ColⅠ） were detected. Metabolomics was analyzed on mice's liver tissues. The mice's serum was collected for metabolomics analysis. ResultsCompared with the model group， Zuojinwan significantly improved indicators related to liver fibrosis and liver injury. Compared with the normal group， the model group showed significantly elevated levels of fibrosis markers such as laminin （LN）， hyaluronic acid （HA）， procollagen typeⅢ （PC-Ⅲ）， and type Ⅳ Collagen （Ⅳ-C）， while liver injury indicators such as alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， alkaline phosphatase （ALP）， lactate dehydrogenase （LDH）， and total bilirubin （TBIL）， exhibited a marked upward trend （P<0.05）. Compared with the model group， the silymarin group showed a significant decrease in the aforementioned indicators （P<0.05）. Notably， compared with the model group， the Zuojinwan group exhibited a significant reduction in all these indicators （P<0.05）， with efficacy comparable to that of the silymarin group. Zuojinwan reduced mRNA and protein levels of α-SMA and ColⅠ in the liver tissue. Metabolomics results revealed that compared with the model group， Zuojiinwan significantly reduced levels of glucose metabolism-related metabolites such as D-fructose 1，6-bisphosphate （FBP）， nicotinamide adenine dinucleotide phosphate （NADPH）， sodium beta-D-fructose 6-phosphate （F6P）， dihydroxyacetone phosphate （DHAP）， fumaric acid， and D-glucose 6-phosphate （G6P） （P<0.05）. Serum enzyme-linked immunosorbent assay （ELISA） was used to detect glucose metabolism indicators and further validate the regulatory effect of Zuojinwan on glucose metabolism. ConclusionThese results suggest that Zuojinwan may improve liver fibrosis by regulating the dysregulated levels of glucose metabolism during the progression of liver fibrosis.

Based on the theory that Shaoyang （少阳） and Shaoyin （少阴） function as the principal pivots， and integrating the laws of qi-fire transformation， yin-yang alternation， and the waxing and waning of healthy qi and pathogenic factors， as well as the action characteristics of immune checkpoint inhibitors （ICIs）， this study systematically expounds the dynamic evolution laws of the Shaoyang and Shaoyin pivots and constructs a stage- and phase-based differentiation and treatment framework for gastric cancer immunotherapy. In the neoadjuvant treatment stage， the core pathogenesis is the dysfunction of the Shaoyang pivot and the disorder of qi and fire transformation， often accompanied by stagnation of cold-deficiency in the middle jiao （焦）. In the postoperative adjuvant treatment stage， the dominant factors are the depletion of Shaoyin fire and the damage of yin fluid， accompanied by the lingering of residual pathogen in the Shaoyang level and the obstruction of collaterals by phlegm and stasis. In the advanced stage， the critical conditions are the exhaustion of the Shaoyin pivot， the blockage of orifices by turbid toxins， and the separation of yin and yang， leading to the rampant of pathogenic toxins and the emergence of various complicated syndromes. The treatment should be guided by the principle of regulating the double pivot. In the neoadjuvant treatment stage， it is recommended to activate the Shaoyang pivot and warm and promote the qi movement in the middle jiao. In the postoperative adjuvant treatment stage， clearing the residual pathogenic factors from the pivot and nourishing the Shaoyin. In the advanced stage， rescuing the Shaoyin pivot and opening the orifices to transform turbidity. Furthermore， stage-based treatment should serve as the foundation for dynamically assessing the patient's immune status and phase-specific changes in immunity， thereby promoting immune activation while preventing and managing immune overactivation and drug resistance.

BACKGROUND:The stress effect of autophagy on epithelial cells,fibroblasts and myofibroblasts is closely related to the formation process of pulmonary fibrosis.OBJECTIVE:To screen the genes related to autophagy in patients with pulmonary fibrosis,and explore their correlation with the prognosis of patients with pulmonary fibrosis,in order to provide a new target for clinical intervention in pulmonary fibrosis.METHODS:The gene expression profiling dataset downloaded from GSE70866 was used as a training set,differentially expressed genes between pulmonary fibrosis patients and normal healthy individuals was analyzed using the R language and intersected with autophagy-related genes to identify the differentially expressed genes with the most significant changes.Multiple analysis methods were used to identify key prognostic genes and construct genetic prognostic models.Patients with pulmonary fibrosis were divided into high-risk and low-risk groups according to their risk scores,and the validity of the prognostic model was verified using the Siena cohort and Leuven cohort validation sets.A cell model of pulmonary fibrosis was established by inducing HFL-1 cells(human embryonic lung fibroblasts)with transforming growth factor-β1,and an animal model of pulmonary fibrosis was established in mice by tracheal instillation of bleomycin to validate the expressions of prognostic genes.RESULTS AND CONCLUSION:(1)There were 2 650 differentially expressed genes between fibrotic tissue and normal tissue.Among them,34 genes related to autophagy showed significant expression changes.(2)Kaplan-Meier survival analysis curves for the Siena cohort and Leuven cohort validation sets showed significantly lower survival in the high-risk group than in the low-risk group.(3)Three autophagy genes related to prognosis were screened out:myelocytomatosis viral oncogene(MYC),C-C motif chemokine ligand 2(CCL2),and GABA type a receptor associated protein like 1(GABARAPL1).(4)Both in vivo and in vitro studies showed that compared with the control group,the expression levels of myelocytomatosis viral oncogene and C-C motif chemokine ligand 2 mRNA and protein were significantly higher in the lung fibrosis model group(P＜0.01,P＜0.05),while the expression levels of GABA type a receptor associated protein like 1 mRNA and protein were lower(P＜0.001).To conclude,bioinformatics methods are used to analyze the expression of three autophagy-related genes in pulmonary fibrosis and their correlation with the prognosis of patients with pulmonary fibrosis.The constructed prognostic model has good predictive ability for the 1-,2-,and 3-year survival rates of patients with pulmonary fibrosis.Moreover,in vivo and in vitro models have been used to verify that myelocytomatosis viral oncogene and C-C motif chemokine ligand 2 are highly expressed in lung fibroblasts and tissues,and that GABA type a receptor associated protein like 1 is lowly expressed.

BACKGROUND:The stress effect of autophagy on epithelial cells,fibroblasts and myofibroblasts is closely related to the formation process of pulmonary fibrosis.OBJECTIVE:To screen the genes related to autophagy in patients with pulmonary fibrosis,and explore their correlation with the prognosis of patients with pulmonary fibrosis,in order to provide a new target for clinical intervention in pulmonary fibrosis.METHODS:The gene expression profiling dataset downloaded from GSE70866 was used as a training set,differentially expressed genes between pulmonary fibrosis patients and normal healthy individuals was analyzed using the R language and intersected with autophagy-related genes to identify the differentially expressed genes with the most significant changes.Multiple analysis methods were used to identify key prognostic genes and construct genetic prognostic models.Patients with pulmonary fibrosis were divided into high-risk and low-risk groups according to their risk scores,and the validity of the prognostic model was verified using the Siena cohort and Leuven cohort validation sets.A cell model of pulmonary fibrosis was established by inducing HFL-1 cells(human embryonic lung fibroblasts)with transforming growth factor-β1,and an animal model of pulmonary fibrosis was established in mice by tracheal instillation of bleomycin to validate the expressions of prognostic genes.RESULTS AND CONCLUSION:(1)There were 2 650 differentially expressed genes between fibrotic tissue and normal tissue.Among them,34 genes related to autophagy showed significant expression changes.(2)Kaplan-Meier survival analysis curves for the Siena cohort and Leuven cohort validation sets showed significantly lower survival in the high-risk group than in the low-risk group.(3)Three autophagy genes related to prognosis were screened out:myelocytomatosis viral oncogene(MYC),C-C motif chemokine ligand 2(CCL2),and GABA type a receptor associated protein like 1(GABARAPL1).(4)Both in vivo and in vitro studies showed that compared with the control group,the expression levels of myelocytomatosis viral oncogene and C-C motif chemokine ligand 2 mRNA and protein were significantly higher in the lung fibrosis model group(P＜0.01,P＜0.05),while the expression levels of GABA type a receptor associated protein like 1 mRNA and protein were lower(P＜0.001).To conclude,bioinformatics methods are used to analyze the expression of three autophagy-related genes in pulmonary fibrosis and their correlation with the prognosis of patients with pulmonary fibrosis.The constructed prognostic model has good predictive ability for the 1-,2-,and 3-year survival rates of patients with pulmonary fibrosis.Moreover,in vivo and in vitro models have been used to verify that myelocytomatosis viral oncogene and C-C motif chemokine ligand 2 are highly expressed in lung fibroblasts and tissues,and that GABA type a receptor associated protein like 1 is lowly expressed.

Chronic heart failure （CHF） is the terminal stage of various heart diseases， with high morbidity， mortality， and hospitalization rate. According to the theory of collateral diseases， the core pathogenesis of CHF is blood stasis of collaterals， water retention and stagnation， and pathogen accumulation in collaterals. Accordingly， the treatment should focus on reinforcing healthy Qi to warm Yang， activating blood to dredge collaterals， and excreting water to alleviate edema. On this basis， Qili Qiangxin capsules （QLQX） are created. This prescription can effectively treat chronic heart failure. Modern studies have shown that QLQX contains a variety of pharmacological components such as flavonoids， alkaloids， terpenoids， phenylpropanoids， phenolic acids， and cardiac glycosides. QLQX can improve the cardiac function， inhibit myocardial fibrosis， improve hemodynamics， mitigate inflammation， reduce cardiomyocyte apoptosis， and regulate the nervous system， with mild adverse reactions. This study analyzed the etiology and pathogenesis of CHF based on the theory of collateral diseases， explored the relationship between the prescription and syndrome， and delved into the material basis and mechanism of QLQX in the treatment of CHF， aiming to provide reference for the clinical application and scientific research of QLQX.

OBJECTIVES: Keloids are fibrotic skin disorders characterized by excessive collagen deposition and a high recurrence rate, closely associated with inflammatory mediators. However, existing epidemiological studies are limited by confounding factors and reverse causality, making it difficult to establish causation. This study aims to investigate the causal relationship between circulating cytokines and keloids using Mendelian randomization analysis. METHODS: Significant single nucleotide polymorphisms (SNPs) associated with circulating cytokines (exposures) and keloids (outcomes) were extracted from genome-wide association study (GWAS) summary datasets. Eligible SNPs were selected as instrumental variables (IVs). Exposure data were derived from a cytokine GWAS including 8 293 Finnish participants, and outcome data from a keloid GWAS based on the UK Biobank. The inverse-variance weighted (IVW) method served as the primary analytical approach to estimate causal effects, supplemented by weighted median (WME), MR-Egger regression, and other sensitivity analyses. Horizontal pleiotropy was assessed using MR-Egger regression and the MR pleiotropy residual sum and outlier (MR-PRESSO) test, while Cochran's Q test evaluated heterogeneity. Leave-one-out analysis was used to verify robustness and consistency. A reverse MR analysis was also conducted, with keloid as the exposure and cytokines as outcomes, to rule out reverse causation. RESULTS: IVW analysis identified significant positive causal associations between two cytokines and keloids-macrophage migration inhibitory factor (MIF) [odds ratio (OR)=2.081, 95% confidence interval (CI) 1.219 to 3.552, P=0.007] and monocyte chemoattractant protein-1 (MCP-1) (OR=1.673, 95% CI 1.036 to 2.701, P=0.035). Conversely, stem cell factor (SCF) showed a negative causal relationship with keloids (OR=0.518, 95% CI 0.269 to 0.998, P=0.049). Results from the MR-Egger and weighted median analyses were consistent with IVW findings. No evidence of horizontal pleiotropy was observed (P>0.05). Except for interleukin-6 (P=0.014), no heterogeneity was detected in other cytokines. Leave-one-out analysis further confirmed the robustness of the causal associations. In reverse MR analysis, keloids were causally related only to β-nerve growth factor (beta-NGF) (OR=1.048, 95% CI 1.002 to 1.095, P=0.039), with no heterogeneity or pleiotropy detected in most cytokines (P>0.05). CONCLUSIONS MIF and MCP-1 exhibit positive causal associations with keloid formation, while SCF shows a negative causal relationship. These findings provide new evidence for the causal involvement of inflammatory cytokines in keloid pathogenesis and offer potential molecular targets for developing novel keloid therapies.
Humans ; Keloid/blood* ; Mendelian Randomization Analysis ; Cytokines/genetics* ; Polymorphism, Single Nucleotide ; Genome-Wide Association Study ; Chemokine CCL2/genetics* ; Interleukin-6/genetics* ; Macrophage Migration-Inhibitory Factors/genetics* ; Male ; Stem Cell Factor/blood* ; Female ; Intramolecular Oxidoreductases

Hepatic ischemia-reperfusion injury (HIRI) has been considered as an inevitable process of liver transplantation. Hepatocyte ferroptosis is a key factor in HIRI development, yet precise mechanism and potential therapies are still unclear. Here, we demonstrated a strong correlation between hepatocyte ferroptosis and the downregulation of poly(rC)-binding protein (PCBP2), which compromised the stability of antiporter system Xc^- (consisted of SL3A2/SLC7A11). Besides, inhibiting PCBP2 contributed to facilitating cofactor p300 to enhance the transcriptional activity of HIF1α, leading to the expression and secretion of HMGB1. Then, released HMGB1 from ferroptotic hepatocytes worsened M1 macrophage recruitment and immune response during HIRI. Additionally, acteoside (ACT) was shown to assist PCBP2 in stabilizing the mRNA stability of Slc3a2 and Slc7a11, as well as enhance the binding affinity of PCBP2-system Xc^-. Beyond that, ACT also supported PCBP2 to limit HMGB1-induced M1 macrophage recruitment through imposing restrictions on p300 and HIF1α. Furthermore, specific knockdown of PCBP2 in hepatocytes directly interposed the therapeutic efficacy of ACT on HIRI mice. In conclusion, ACT alleviated hepatocyte ferroptosis and HIRI via promoting PCBP2 to maintain the stability of system Xc^- and limit HIF1α/p300-HMGB1 signaling. These findings highlight the therapeutic benefits of ACT in treating HIRI and offer insights into innovative therapeutic strategies.

The identification and optimization of mutations in nanobodies are crucial for enhancing their therapeutic potential in disease prevention and control. However, this process is often complex and time-consuming, which limit its widespread application in practice. In this study, we developed a workflow, named Evolutionary-Nanobody (EvoNB), to predict key mutation sites of nanobodies by combining protein language models (PLMs) and molecular dynamic (MD) simulations. By fine-tuning the ESM2 model on a large-scale nanobody dataset, the ability of EvoNB to capture specific sequence features of nanobodies was significantly enhanced. The fine-tuned EvoNB model demonstrated higher predictive accuracy in the conserved framework and highly variable complementarity-determining regions of nanobodies. Additionally, we selected four widely representative nanobody-antigen complexes to verify the predicted effects of mutations. MD simulations analyzed the energy changes caused by these mutations to predict their impact on binding affinity to the targets. The results showed that multiple mutations screened by EvoNB significantly enhanced the binding affinity between nanobody and its target, further validating the potential of this workflow for designing and optimizing nanobody mutations. Additionally, sequence-based predictions are generally less dependent on structural absence, allowing them to be more easily integrated with tools for structural predictions, such as AlphaFold 3. Through mutation prediction and systematic analysis of key sites, we can quickly predict the most promising variants for experimental validation without relying on traditional evolutionary or selection processes. The EvoNB workflow provides an effective tool for the rapid optimization of nanobodies and facilitates the application of PLMs in the biomedical field.

Objective To explore the effects and mechanism of Jianpi Shuyi Decoction in improving pancreatic fibrosis in chronic pancreatitis(CP)based on network pharmacology and animal experiments.Methods TCMSP was used to screen the active components and targets of Jianpi Shuyi Decoction.GeneCards was used to obtain the disease targets of pancreatic fibrosis,and the intersection of drug and disease targets was used to construct the protein interaction network and the drug-component-target network,and the core target genes were screened out.GO and KEGG pathway enrichment analysis was performed on the intersecting targets.Caerulein was used to induce CP mouse model,and Jianpi Shuyi Decoction was given for gavage.HE and Sirius red staining were used to observe pancreatic tissue inflammation and collagen deposition,respectively.RT-qPCR was used to observe the mRNA expression levels of Acta2,COL1A1,PI3K and Akt1 in pancreatic tissue.Immunohistochemistry staining was used to observe the protein expression levels of α-SMA,COL-1,p-PI3K and p-Akt in pancreatic tissues.Results A total of 181 active components were screened from Jianpi Shuyi Decoction,corresponding to 284 targets,with 240 targets overlapping between drugs and disease and the core targets were PTGS2,HSP90AA1,etc.193 signaling pathways were obtained from KEGG pathway enrichment analysis,primarily involving lipids and atherosclerosis,chemical carcinogenic-receptor activation,PI3K-Akt signaling pathway and others.The results of animal experiments showed that,compared with the normal group,the model group showed a large number of inflammatory cell infiltration and collagen deposition in pancreatic tissue,the mRNA expression of Acta2,COL1A1,PI3K and Akt1 in pancreatic tissue significantly increased(P<0.01),and the protein expression of α-SMA,COL-1,p-PI3K,p-Akt significantly increased(P<0.01);Jianpi Shuyi Decoction significantly reduced the inflammation and collagen deposition in pancreas of mice,reduced the mRNA expression of Acta2,COL1A1,PI3K and Akt1(P<0.05),and attenuated the protein expression of α-SMA,COL-1,p-PI3K and p-Akt in pancreatic tissue(P<0.05).Conclusion Jianpi Shuyi Decoction may exert a therapeutic effect on CP pancreatic fibrosis by regulating the PI3K/Akt signaling pathway,attenuating inflammation and collagen deposition in the pancreas,and reducing the levels of α-SMA and COL-1.

Cancer incidence in China has been rising steadily,with a particularly heavy burden from several high-prevalence malignancies.Medical examination for cancer plays a critical role in the early detection of cancer,precancerous lesions,and precursor conditions,thereby facilitating timely diagnosis and intervention.Such examination also addresses the growing demand for person-alized cancer screening services among diverse population groups.The development of evidence-based,context-specific cancer screening guidelines is essential to enhance the standardization,quality,and equity of preventive screening practices across the country,ultimately improving out-comes in early cancer detection and treatment.Guided by the Department of Medical Emergency Response of the National Health Commission,the Guidelines for Medical Examination for Cancer in Health Examination Agency(2025 Edition)were developed under the leadership of the National Cancer Center.A multidisciplinary panel of experts formulated the guidelines in accordance with the principles and methodology of the World Health Organization Handbook for Guideline Deve-lopment.The guidelines provide evidence-based recommendations on key clinical domains:target cancers and populations,overall screening workflow,screening protocols,diagnostic technolo-gies,result interpretation,follow-up procedures,and quality control.The primary objective is to standardize cancer screening practices in health examination agency and strengthen China's ca-pacity for prevention and control of high-burden cancers.