Objective To explore the feasibility and reference value of allogeneic lung transplantation and postoperative monitoring in miniature pigs for lung transplantation research. Methods Two miniature pigs (R1 and R2) underwent left lung allogeneic transplantation. Complement-dependent cytotoxicity tests and blood cross-matching were performed before surgery. The main operative times and partial pressure of arterial oxygen (PaO₂) after opening the pulmonary artery were recorded during surgery. Postoperatively, routine blood tests, biochemical blood indicators and inflammatory factors were detected, and pathological examinations of multiple organs were conducted. Results The complement-dependent cytotoxicity test showed that the survival rate of lymphocytes between donors and recipients was 42.5%-47.3%, and no agglutination reaction occurred in the cross-matching. The first warm ischemia times of D1 and D2 were 17 min and 10 min, respectively, and the cold ischemia times were 246 min and 216 min, respectively. Ultimately, R1 and R2 survived for 1.5 h and 104 h, respectively. Postoperatively, in R1, albumin (ALB) and globulin (GLB) decreased, and alanine aminotransferase increased; in R2, ALB, GLB and aspartate aminotransferase all increased. Urea nitrogen and serum creatinine increased in both recipients. Pathological results showed that in R1, the transplanted lung had partial consolidation with inflammatory cell infiltration, and multiple organs were congested and damaged. In R2, the transplanted lung had severe necrosis with fibrosis, and multiple organs had mild to moderate damage. The expression levels of interleukin-1β and interleukin-6 increased in the transplanted lungs. Conclusions The allogeneic lung transplantation model in miniature pigs may systematically evaluate immunological compatibility, intraoperative function and postoperative organ damage. The data obtained may provide technical references for subsequent lung transplantation research.

Metabolic dysfunction-associated fatty liver disease （MAFLD） has become one of the most prevalent chronic liver diseases worldwide， posing a serious challenge to public health. In this context， the integration of artificial intelligence （AI）， especially intelligent diagnosis and treatment and digital health interventions based on machine learning， can break through the limitations of traditional methods， realize efficient screening of multi-dimensional data such as key genes， biomarkers， and biochemical metabolites， and achieve revolutionary breakthroughs in risk prediction， subtype identification， and therapeutic effect assessment for MAFLD. This article systematically reviews the ground-breaking application of machine learning models in driving the innovation of clinical diagnosis and precise risk prediction of MAFLD， conducts a comprehensive comparative analysis of digital health practice cases of MAFLD in China and globally， and deeply analyzes their advantages and limitations in terms of research subjects， interventions， and management team. Studies have shown that the deep integration of digital health and long-term management of MAFLD is becoming the key engine driving the transformation of disease management modes towards an intelligent， individualized， and precise era， but there are various ethical and technical issues that need to be addressed urgently.

Background/Aims: Lenvatinib resistance remains a critical barrier in advanced hepatocellular carcinoma (HCC) therapy. However, the underlying mechanisms and strategies for reversing resistance remain incompletely understood. Methods: Integrated transcriptomics of lenvatinib-resistant patient tumors and an acquired-resistance murine model identified a novel macrophage subpopulation. Functional validation employed CRISPR-SAM screening, conditioned medium (CM) assays, subcutaneous/orthotopic xenografts, patient-derived organoids (PDOs), and patient-derived xenografts (PDXs). Mechanistic studies included ChIP-qPCR, co-immunoprecipitation, and pharmacologic targeting. Clinical relevance was assessed in a retrospective cohort. Results: Resistant HCC exhibited significant enrichment of a COLEC12high TAM subset , which correlated with poor survival and treatment response. These TAMs secreted neuregulin-1 (NRG1) , activating HER2/HER3-AKT signaling in tumor cells to drive cancer stemness and lenvatinib resistance. Mechanistically, in TAMs COLEC12 sequestered STAT1 in the cytoplasm, preventing its phosphorylation, and thereby derepressing STAT3-mediated NRG1 transcription. Depletion of NRG1 reversed the stemness phenotypes and resensitized tumors to lenvatinib both in vitro and in vivo. Clinically, high NRG1 expression predicted an inferior lenvatinib response and shorter survival. Crucially, the bispecific anti-HER2/HER3 antibody zenocutuzumab restored lenvatinib efficacy in PDOs, PDXs, and murine models. Conclusions Our work establishes the COLEC12high TAM/NRG1 axis as a master regulator of therapeutic resistance and identifies NRG1 as a predictive biomarker, providing a clinically actionable strategy to overcome lenvatinib resistance in HCC.

Objective: To analyze and compare the levels of various coagulation factors in cryoprecipitate and corresponding fresh frozen plasma, thereby providing a basis for the precise clinical administration of cryoprecipitate. Methods: Fibrinogen, coagulation factors Ⅱ, Ⅴ, Ⅶ, Ⅷ, Ⅸ, Ⅹ, Ⅺ, Ⅻ, and ⅩⅢ, von Willebrand factor antigen (VWF∶Ag), and von Willebrand factor activity (VWF∶Act) were measured in cryoprecipitate and corresponding fresh frozen plasma prepared from 400 mL of whole blood. The levels of each coagulation factor were quantified and subjected to paired comparisons; the total coagulation factor content was also calculated for each bag of cryoprecipitate and fresh frozen plasma. Results: The fibrinogen level in the precipitate was (4.48±0.83) g/L, coagulation factor Ⅷ activity was (330.11±172.00) IU/dL, VWF∶Ag was (510.7±229.34) IU/dL, VWF∶Act was (341.35±116.21) IU/dL; the activities of other coagulation factors did not differ significantly from those in fresh frozen plasma. In each bag of cryoprecipitate (47.7±1.68) mL, the fibrinogen content was (211.94±39.6) mg, the coagulation factor Ⅷ content was (156.67±79.82) IU, the VWF∶Ag content was (245.18±103.25) IU, VWF∶Act was (164.21±54.30) IU. Paired comparisons between cryoprecipitate and fresh frozen plasma revealed that the concentrations of Fib, FⅧ, FⅨ, FⅪ, FⅫ, FⅩⅢ, VWF∶Ag, and VWF∶Act in cryoprecipitate were all higher than those in fresh frozen plasma, with statistically significant differences (P<0.05). Conclusion: Fresh frozen plasma and cryoprecipitate can serve as treatment options for patients with coagulation factor deficiencies. The results provide data to guide the precise selection of blood products and the rational use of blood in clinical practice.

Objective To target at the NKG2A-HLA-E inhibitory axis,a pluripotent stem cell(iPSC)-derived geneti-cally engineered natural killer cells(NK cells)with NKG2A knockout(NKG2A KO-iNK)were prepared and then their tumor-killing efficacy was evaluated in vitro.Methods NKG2A was knocked out in iPSCs using gene-editing technology.These cells were then differentiated into NKG2A KO-iNK cells.Surface markers at each differentiation stage were analyzed by flow cytometry.Western blot confirmed NKG2A knockout,and flow cytometry assessed expres-sion of activating receptors(NKG2D)and natural cytotoxicity receptors(NKp30,NKp44,NKp46)in NKG2A KO-iNK cells.Cytotoxic activity against tumor cell lines with varying human leukocyte antigen E(HLA-E)expression level was evaluated via lactate dehydrogenase(LDH)release assay.Results Co-transfection of iPSCs with Cas9 pro-tein and three small-guide RNAs(sgRNAs)targeting at exons 1 and 2 of the KLRC1 gene(encoding NKG2A)suc-cessfully generated monoclonal NKG2A-knockout iPSCs(NKG2A KO-iPSCs)with a single T-base insertion in exon 1.During iPSC differentiation into NK cells,CD34 expression reached 30％-50％at the embryoid body(EB)stage(day 8),while CD56 and CD 16 expression exceeded 80％by day 28.Western blot confirmed complete NKG2A knockout in NKG2A KO-iNK cells.Flow cytometry revealed comparable expression level of activating receptor NKG2D and cytotox-icity receptors(NKp30,NKp44,NKp46)between NKG2A KO-iNK and wild-type iNK(WT-iNK)cells.The LDH assay results indicated that the cytotoxic activity of NKG2A KO-iNK cells against the HLA-E highly-expressed B-cell precursor leukemia cell line Nalm6 cells was significantly higher than that of WT-iNK cells,while there was no signif-icant difference between them and human myeloma cell line H929 cells with low HLA-E expression and human hepa-tocellular carcinoma cell line HepG2 cells with almost no HLA-E expression.Interferon-γ(IFN-γ)pretreatment up regulated HLA-E expression in Nalm6 cells,further amplifying NKG2A KO-iNK-mediated cytotoxicity.Conclusions By disrupting the NKG2A-HLA-E inhibitory axis,NKG2A KO-iNK cells exhibit markedly enhanced in vitro cytotoxic-ity against HLA-E-high tumor cells.This result highlights their potential function as a novel adoptive cell therapy strategy for cancers reliant on HLA-E-mediated immune evasion.

Objective To investigate the value of combined prediction of serum 25-hydroxyvita-min D[25-(OH)-D],homocysteine(Hey)and brain natriuretic peptide(BNP)for pregnancy out-comes in patients with hypertensive disorders of pregnancy(HDP).Methods A total of 200 patients with HDP were selected as research subjects and divided into gestational hypertension(GH)group(n=84),mild preeclampsia(PE)group(n=67)and severe PE group(n=49)according to the severity of the disease.Additionally,50 healthy pregnant women were selected as control group.The serum levels of 25-(OH)-D,Hey and BNP were compared among the groups,and their relationships with the severity of HDP were analyzed.According to the pregnancy outcomes of HDP patients,they were divided into poor pregnancy outcome group(n=82)and good pregnancy outcome group(n=118).The serum levels of 25-(OH)-D,Hey and BNP were compared between the poor and good pregnancy outcome groups.Logistic regression analysis was used to screen for the influencing factors of poor pregnancy outcomes in HDP patients.Results Compared with the control group,the severe PE group had the lowest serum 25-(OH)-D level,followed by the mild PE group and the GH group,with statistically significant differences(P＜0.001).Compared with the control group,the severe PE group had the highest serum Hcy and BNP levels,followed by the mild PE group and the GH group,with statistically significant differences(P＜0.001).Serum 25-(OH)-D was negatively cor-related with the severity of HDP in patients(r=-0.427,P＜0.001),while serum Hey and BNP were positively correlated with the severity of HDP in patients(r=0.585 and 0.682;P＜0.001).The serum 25-(OH)-D level in the poor pregnancy outcome group was significantly lower than that in the good pregnancy outcome group,while the serum Hcy and BNP levels were significantly higher than those in the good pregnancy outcome group(P＜0.05).Logistic regression analysis results showed that elevated serum Hey and BNP levels,severe PE and elevated systolic blood pressure were independent risk factors for poor pregnancy outcomes in HDP patients,while high 25-(OH)-D level was a protective factor(P＜0.05).The area under the curve(AUC)of the prediction model constructed based on the Logistic regression analysis results was 0.911(95％CI,0.863 to 0.947),with asensitivity of 91.46％and specificity of 75.42％.Conclusion As the severity of HDP in pa-tients increases,their serum 25-(OH)-D level decreases,while serum Hey and BNP levels in-crease.The combined prediction of serum 25-(OH)-D,Hey and BNP has a high value for predic-ting poor pregnancy outcomes in HDP patients.

Objective To investigate the impact of anticentromere antibody(ACA)on the clinical features and prognosis of patients with primary biliary cholangitis(PBC)by comparing clinical classification,ursodeoxycholic acid(UDCA)response,GLOBE score,and UK-PBC score between ACA-positive PBC patients and ACA-negative PBC patients.Methods A total of 749 patients who were admitted to Beijing Ditan Hospital,Capital Medical University,from August 2013 to December 2022 and were diagnosed with PBC were enrolled and divided into ACA-positive group with 147 patients and ACA-negative group with 602 patients.According to their conditions on admission,the two groups were compared in terms of the distribution of clinical types,i.e.,chronic progression-type PBC,portal hypertension-type PBC,and standard jaundice/liver failure-type PBC.There were 261 patients with complete data after 1-year follow-up,among whom there were 53 patients with positive ACA and 208 with negative ACA.A statistical analysis was performed,and propensity score matching was performed based on sex and age at a ratio of 1∶2.The two groups were compared in terms of 1-year UDCA response rate,GLOBE score,and UK-PBC score before and after matching.The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups,and the chi-square test was used for comparison of categorical data between two groups.Results Compared with the ACA-negative group,the ACA-positive group had a significantly higher age(61.28±10.35 years vs 56.74±12.17 years,t=4.164,P<0.001),a significantly higher proportion of female patients(93.9%vs 77.6%,χ2=20.221,P<0.001),a significantly higher proportion of patients with portal hypertension(48.3%vs 27.6%,χ2=23.289,P<0.001),and a significantly lower proportion of patients with jaundice/liver failure(24.5%vs 38.5%,χ2=10.205,P<0.001).After 1-year follow-up,for the 261 PBC patients with complete data,there was no significant difference in UDCA response rate before propensity score matching between the ACA-positive group and the ACA-negative group(41.5%vs 41.8%,P>0.05),and there was a significant difference in the proportion of patients with a GLOBE score of>0.3 between the ACA-positive group and the ACA-negative group(92.5%vs 80.3%,χ2=3.935,P=0.047).There were 53 patients in the ACA-positive group and 106 patients in the ACA-negative group after propensity score matching,and there were no significant differences between the two groups in UDCA response rate,GLOBE score,and UK-PBC score(all P>0.05).Conclusion ACA-positive patients tend to have an older age,with a higher proportion of female patients or patients with portal hypertension,while there is a relatively low proportion of patients with jaundice/liver failure.Positive ACA has no significant impact on UDCA response rate,GLOBE score,and UK-PBC score.

Objective To observe value of real-time functional MRI neurofeedback(rtfMRI-NF)for modulating brain activity changes in obese adults.Methods Twenty-two obese adults were prospectively recruited,3-week rtfMRI-NF training intervention was conducted.Barratt impulsivityness scale version 11(BIS-11),three-factor eating questionnaire(TFEQ)and food rating scales were scored before and after intervention,and whole-brain resting state fMRI(rs-fMRI)data were obtained.Clinical scale scores,rs-fMRI regional homogeneity(ReHo)values and amplitude of low frequency fluctuation(ALFF)values were compared before and after intervention.Then brain regions with differences of ReHo and ALFF values before and after intervention were explored,and correlations of values of rs-fMRI data differences and clinical scale score differences were analyzed.Results BIS-11 score,TFEQ emotional eating(TFEQ-EE)score and food rating scales scores decreased,while TFEQ cognitive restraint(TFEQ-CR)scores increased in obese adults after intervention(all P＜0.05).ReHo values in right inferior frontal gyrus,right anterior cingulate and left precuneus increased,while of left middle temporal gyrus decreased(all corrected P＜0.05).ALFF values of left precuneus increased,whereas of left middle occipital gyrus,right superior occipital gyrus,left calcarine fissure and surrounding cortex and left supramarginal and angular gyrus decreased(all corrected P＜0.05).ReHo difference in right anterior cingulate was negatively correlated with BIS-11 scores difference(r=-0.601,P＜0.05),and ALFF difference in left precuneus was negatively correlated with TFEQ-EE difference(r=-0.478,P＜0.05).Conclusion rtfMRI-NF intervention could correct disorder of cerebral functional areas in obese adults and change high-calorie food preference and poor dietary habits.

Breast cancer remains one of the most prevalent malignancies among women worldwide,with distant organ metastasis being the leading cause of mortality.The lungs are a common site of metastasis in breast cancer and are associated with significantly poor prognosis.Although notable progress has been made in therapeutic strategies,efficacy remains markedly limited once pulmonary metastasis develops.Therefore,elucidating the molecular mechanisms that drive pulmonary metastasis,characterizing the immune microenvironment,and as-sessing therapeutic responsiveness are critical.Recent advances in multi-omics approaches-including genomics,transcriptomics,proteomics,metabolomics,and epigenetics-have been widely applied to investigate breast cancer metastasis,offering new insights into the complex mo-lecular networks underlying pulmonary dissemination.This review synthesizes current findings on key genes,proteins,metabolites,and epi-genetic regulatory mechanisms involved in the metastatic cascade from a multi-omics perspective.

Objective:To investigate the relationship between nerve growth factor (NGF) concentration in follicular fluid and abnormal menstrual cycle in infertile patients with polycystic ovary syndrome (PCOS).Methods:A retrospective cohort study was conducted on 100 infertile patients with PCOS who underwent in vitro fertilization and embryo transfer (IVF-ET) at Department of Obstetrics and Gynecology, Peking University Third Hospital from March 2017 to June 2019. For comparison, the 100 patients with PCOS were divided into low NGF group ( n=50) and high NGF group ( n=50) based on the median NGF concentration (1 644.03 ng/L) in follicular fluid. Baseline characteristics, menstrual status and clinical outcomes of assisted reproductive technology were compared. We performed multiple linear regression analysis to examine the effect of NGF in follicular fluid on menstrual cycle length for multivariate analysis. Results:1) PCOS patients in the low NGF group had significantly higher body mass index [(27.24±5.17) kg/m 2] and white blood cell count [7.31(5.99, 8.43)×10 9/L ] than those in the high NGF group [(25.03±4.46) kg/m 2, P=0.024; 5.95(5.08,7.01)×10 9/L, P=0.001], while high-density lipoprotein cholesterol [1.15 (0.98, 1.36) mmol/L] and basic follicle-stimulating hormone level [6.51 (5.10,7.95) U/L] in the low NGF group were significantly lower than those in the high NGF group [1.36 (1.09,1.52) mmol/L, P=0.039;6.51 (5.10,7.95)U/L, P=0.040]. 2) PCOS patients in the low NGF group had significantly higher menstrual cycle length [60.00 (35.00, 180.00) d] than the high NGF group [32.50 (27.00,67.50) d, P=0.001]. 3) Multiple linear regression analysis revealed that after adjustment for body mass index, age, infertility duration, infertility type, and glucose and lipid metabolic parameters, the NGF concentration in the follicular fluid independently and negatively correlated with menstrual cycle length ( P<0.05). 4) The NGF concentration in follicular fluid was not correlated with assisted reproductive outcomes. Conclusion:NGF concentration in follicular fluid is closely related to the degree of menstrual cycle abnormalities in patients with PCOS.