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.

BACKGROUND: The level of measurable residual disease (MRD) before and after transplantation is related to inferior transplant outcomes, and post-hematopoietic stem cell transplantation measurable residual disease (post-HSCT MRD) has higher prognostic value in determining risk than pre-hematopoietic stem cell transplantation measurable residual disease (pre-HSCT MRD). However, only a few work has been devoted to the risk factors for positive post-HSCT MRD in patients with acute lymphoblastic leukemia (ALL). This study evaluated the risk factors for post-HSCT MRD positivity in patients with ALL who underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: A total of 1683 ALL patients from Peking University People's Hospital between January 2009 and December 2019 were enrolled to evaluate the cumulative incidence of post-HSCT MRD. Cox proportional hazard regression models were built for time-to-event outcomes. Multivariable analysis was performed to determine independent influencing factors from the univariable analysis. RESULTS: Both in total patients and in T-cell ALL or B-cell ALL, pediatric or adult, human leukocyte antigen-matched sibling donor transplantation or haploidentical SCT subgroups, positive pre-HSCT MRD was a risk factor for post-HSCT MRD positivity ( P  <0.001 for all). Disease status (complete remission 1 [CR1] vs . ≥CR2) was also a risk factor for post-HSCT MRD positivity in all patients and in the B cell-ALL, pediatric, or haploidentical SCT subgroups ( P  = 0.027; P  = 0.003; P  = 0.035; P  = 0.003, respectively). A risk score for post-HSCT MRD positivity was developed using the variables pre-HSCT MRD and disease status. The cumulative incidence of post-HSCT MRD positivity was 12.3%, 25.1%, and 38.8% for subjects with scores of 0, 1, and 2-3, respectively ( P  <0.001). Multivariable analysis confirmed the association of the risk score with the cumulative incidence of post-HSCT MRD positivity and relapse as well as leukemia-free survival and overall survival. CONCLUSION Our results indicated that positive pre-MRD and disease status were two independent risk factors for post-HSCT MRD positivity in patients with ALL who underwent allo-HSCT.
Humans ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/pathology* ; Neoplasm, Residual ; Hematopoietic Stem Cell Transplantation/methods* ; Male ; Female ; Risk Factors ; Adolescent ; Adult ; Child ; Child, Preschool ; Young Adult ; Middle Aged ; Infant ; Transplantation, Homologous ; Proportional Hazards Models ; Retrospective Studies

AIM:To investigate the impact of recombinant human CC16 protein(rhCC16)on cigarette smoke extract(CSE)-induced senescence-associated secretory phenotype(SASP)in human bronchial epithelial cells(HBECs)and in the lung tissues of chronic obstructive pulmonary disease(COPD)mice,and to explore the underlying mechanism.METHODS:HBECs were induced into cellular senescence using 5%CSE.The senescent HBECs were treated with 250 ng/mL rhCC16,and the levels of reactive oxygen species(ROS)were assessed using the 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)method.The levels of trimethylated histone H3 at lysine 9(H3K9me3),a marker of senescence-associated heterochromatic foci(SAHF),were detected using a Western blot assay.RT-qPCR and ELISA were utilized to measure the mRNA expression and protein levels of SASP components including interleukin-1 beta(IL-1β),IL-6,IL-8,chemokine(C-X-C motif)ligand-1(CXCL-1),matrix metalloproteinase 1(MMP1)and MMP3.Passive smoking was con-ducted for six months to induce COPD in mice.RhCC16(2.5 μg/g body weight)or an equal volume of PBS(20 μL)was intranasally administered from the 16th week of smoking in the COPD+rhCC16 group or COPD+PBS group,respectively,with administration 2 hours before smoking.ROS levels in lung tissue cells were investigated using DCFH-DA staining.H3K9me3 levels in lung tissues were tested using Western blot assay.RT-qPCR and ELISA were performed to examine the mRNA expression and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3.RESULTS:DCFH-DA staining results showed that CSE stimulation increased ROS levels in HBECs,while rhCC16 treatment reduced them(P＜0.01).Western blot results indicated that CSE stimulation elevated H3K9me3 levels in HBECs,which were decreased with rhCC16 treatment(P＜0.01).RT-qPCR and ELISA assays demonstrated that CSE stimulation upregulated the mRNA and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3 in HBECs,which were reduced with rhCC16 admin-istration(P＜0.05).DCFH-DA staining results showed an increase in ROS levels in the lung tissues of COPD mice,which were decreased with rhCC16 administration(P＜0.01).Western blot data revealed an increase in H3K9me3 levels in the lung tissues of COPD mice,which were reduced with rhCC16 treatment(P＜0.01).RT-qPCR and ELISA assays demon-strated an upregulation of the mRNA and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3 in the lung tis-sues of COPD mice,which were reduced with rhCC16 treatment(P＜0.05).No statistically significant differences were ob-served in the above-mentioned indicators between the lung tissues of COPD and COPD+PBS mice(P＞0.05).CONCLU-SION:rhCC16 can effectively inhibit CSE-induced SASP in HBECs and in the lung tissues of COPD mice,with its under-lying mechanism potentially related to the inhibition of the ROS-H3K9me3 signaling pathway.

AIM:To investigate autophagic status in ischemic stroke with Liver Yang Hyperactivity and the mechanism of Tianma Gouteng Decoction(TMGTD).METHODS:SD rats were divided into sham,model,TMGTD high/medium/low-dose(20.52/10.26/5.13 g·kg-1·d-1),and Nimodipine(30 mg·kg-1·d-1)groups.A Liver Yang Hyperactivity and cerebral ischemia-reperfusion model was es-tablished using Fuzi Decoction(2 g·kg-1·d-1)and thread-occlusion.After 21 days of Fuzi decoction pretreatment,rats received daily drug administra-tion for 12 days.Syndrome indicators(irritability,24-hour water intake,24-hour urine volume,facial temperature)were recorded,plasma NE,E,cAMP,and cGMP were measured by ELISA,neurological function was assessed using Zea Longa and mNSS methods,brain histopathology was evaluated by HE staining,protein expression of soluble/insoluble p62 and LC3B was detected by Western blot,au-tophagy-related genes were analyzed by PCR array,additionally,mRNA and protein levels of CXCR4 and CXCL12 were measured by qRT-PCR and Western blot.RESULTS:Compared to the sham group,the model group showed increased irritability,24-hours water intake,24-hours urine volume,facial temper-ature,and level of NE,E,cGMP(P＜0.01),neurologi-cal scores(P＜0.01),LC3B-Ⅱ,insoluble p62,CXCR4,CXCL12 expression(P＜0.01),but decreased soluble p62(P＜0.01).TMGTD groups exhibited reduced irri-tability,water intake,urine volume,facial tempera-ture,NE,E,cGMP(P＜0.05,P＜0.01),neurological scores(P＜0.05,P＜0.01),p62 expression(P＜0.01),alongside increased LC3B-Ⅱ(P＜0.01)and improved cortical pathology.TMGD also reversed dysregulat-ed autophagy-apoptosis genes(CXCR4,Lamp1,Tgfb1,APP,Rab24)and reduced CXCR4,CXCL12 ex-pression(P＜0.01).CONCLUSION:In the Liver Yang Hyperactivity and cerebral ischemia-reperfusion model,autophagy genes were activated but flux was impaired,and Tianma Gouteng Decoction may protect by restoring autophagic flux and inhibiting the CXCL12/CXCR4 axis.

Objective:To explore the impact of the two-way referral model on compliance and prognosis in patients with heart failure.Methods:This bidirectional cohort study enrolled chronic heart failure (CHF) patients treated at Qilu Hospital of Shandong University or designated primary hospitals between March 2018 and March 2022. Patients were categorized into two groups based on referral status: two-way referral group (participating in the referral model with≥1 follow-up visit at primary hospitals) and the core hospital group (receiving treatment and follow-up exclusively at Qilu Hospital). Baseline clinical characteristics were collected and compared between groups. Patients underwent followed-up, with primary endpoints including follow-up rate, drug (β-blockers, angiotension converting enzyme inhibitor (ACEI)/angiotensin Ⅱ receptor blockers (ARB)/angiotensin receptor-neprilysin inhibitor (ARNI), sodium-glucose cotransporter 2 inhibitors and mineralocorticoid receptor antagonists) utilization rate and target dose achievement rate. Secondary endpoints encompassed changes from baseline in left ventricular ejection fraction (LVEF), left ventricular end-diastolic diameter (LVEDd), and N-terminal pro-brain natriuretic peptide (NT-proBNP), plus cardiovascular mortality and heart failure rehospitalization. Generalized linear mixed models analyzed longitudinal trends in LVEF, LVEDd, and NT-proBNP levels. Kaplan-Meier curves and Cox regression evaluated LVEF recovery rates, supplemented by subgroup analyses. Multivariate logistic regression was used to identify factors influencing target dose achievement rate for β-blockers and ACEI/ARB/ARNI therapies in CHF patients.Results:A total of 357 patients were enrolled, aged 53 (41, 63) years, including 256 males (71.7%). 157 patients were in the two-way referral group and 200 patients in the core hospital-treated group. Compared with the core hospital-treated group, the two-way referral group had lower baseline LVEF (28 (22, 34)% vs. 31 (23, 36)%, P=0.021) and systolic blood pressure (116 (104, 125) mmHg vs. 121 (109, 134) mmHg (1 mmHg=0.133 kPa), P=0.010). The 12-month follow-up rate of the two-way referral group was higher than the core hospital-treated group (73.8% vs. 56.0%, P=0.004). No significant between-group differences were observed in drug utilization rate of β-blockers, ACEI/ARB/ARNI, or sodium-glucose cotransporter 2 inhibitors during follow-up (all P>0.05), while mineralocorticoid receptor antagonists use showed a declining trend in both groups. Although the core hospital-treated group had higher target dose achievement rates for β-blockers (65.4% vs. 49.3%, P=0.042) and ACEI/ARB/ARNI (79.8% vs. 65.8%, P=0.046) than the two-way referral group, multivariate logistic regression indicated that the two-way referral model was not a negative predictor for these outcomes (all P>0.05). Both groups showed improved NT-proBNP, LVEDd, and LVEF from baseline (all P<0.001) with no significant difference in trends between groups (all P>0.05). There was no significant difference in the composite incidence (7.6% vs. 6.5%, P=0.674) and cumulative incidence (log-rank P=0.684) of cardiovascular death and heart failure rehospitalization at 12 months between two groups. Conclusion:The two-way referral model demonstrates advantages in improving medication adherence, drug utilization rates, and targetdoseachievement rates among CHF patients. This model not only promotes cardiac functional recovery but also reduces risks of cardiovascular mortality and heart failure rehospitalization, achieving comparable therapeutic and management outcomes to those observed in core hospital-treated patients.

Objective:To evaluate the clinical value of holographic imaging-based digital localization technology in robot-assisted partial nephrectomy（RAPN）for the treatment of completely endophytic renal tumors.Methods:A retrospective analysis was conducted on the clinical data of 23 patients with completely endophytic renal tumors who underwent RAPN at the First Affiliated Hospital of Xiamen University between December 2022 and December 2024. Patients were divided into two groups based on the use of holographic imaging：the holographic imaging group（16 cases）and the conventional group（7 cases）. There was no statistically significant difference between the holographic imaging group and the conventional group in terms of age［（41.9 ± 13.4）years vs.（46.9 ± 13.4）years］，body mass index［（25.6 ± 4.8）kg/m2 vs.（24.7 ± 3.1）kg/m2］，maximum tumor diameter［（3.1 ± 0.9）cm vs.（3.0 ± 9.0）cm］，tumor volume［（13.2 ± 9.0）cm3 vs.（34.9 ± 9.9）cm3］，R.E.N.A.L. score［（9.4 ± 1.2）points vs.（9.9 ± 0.7）points］，PADUA score［（10.4 ± 0.7）points vs.（9.4 ± 0.7）points］，proportion of T 1a stage patients［12 cases（75.0%）vs. 6 cases（85.7%）］and preoperative serum creatinine［（67.4 ± 9.5）μmol/L vs.（78.0 ± 16.0）μmol/L］. In the holographic group，holographic models were reconstructed based on preoperative enhanced CT or MRI images and used for preoperative planning and intraoperative localization. In the conventional group，surgeons relied on preoperative CT or MRI images for cognitive fusion during RAPN. Perioperative parameters such as warm ischemia time，operative time，tumor localization time，positive surgical margin rate，and renal function were compared between the two groups. Results:The operative time in the holographic group was significantly shorter than that in the conventional group［（152.8 ± 12.9）min vs.（218.4 ± 105.5）min， P = 0.001］. Warm ischemia time［（26.9 ± 3.4）min vs.（30.7 ± 3.8）min， P < 0.001］，localization time［（4.2 ± 0.9）min vs.（8.9 ± 1.7）min， P < 0.001］，and estimated blood loss［（47.0 ± 17.7）ml vs.（128.6 ± 87.8）ml， P < 0.001］were also significantly lower in the holographic group. In the conventional group，one patient underwent radical nephrectomy，while no patient in the holographic imaging group required conversion to radical nephrectomy. No cases of positive surgical margins were identified in either group. Serum creatinine levels measured one month after surgery showed no statistically significant differences between the two groups［（79.5 ± 15.7）μmol /L vs.（104.9 ± 22.5）μmol /L］. Conclusions:The application of holographic imaging-based digital localization technology in RAPN for completely endophytic renal tumors significantly reduces operative time，localization time，warm ischemia time，and intraoperative blood loss. This technology improves surgical efficiency and success rates，offering distinct clinical advantages.

Background and Aims:Pancreatic cancer(PC)is a highly lethal gastrointestinal malignancy with poorly understood pathogenesis.Previous studies suggest that alterations in plasma metabolomics may be associated with PC development;however,traditional observational studies are prone to confounding and reverse causation,making it difficult to establish causal relationships.This study employed a two-sample Mendelian randomization(MR)approach to systematically evaluate the potential causal relationship between 325 nuclear magnetic resonance(NMR)metabolites and PC risk.Methods:Genome-wide association study(GWAS)data of 325 NMR metabolites from the UK Biobank were integrated with GWAS data of PC from FinnGen.Single nucleotide polymorphisms(SNPs)significantly associated with metabolites were selected as instrumental variables.The inverse variance weighted method served as the primary analysis,supplemented by MR-Egger regression,weighted median,weighted mode,Bayesian weighted Mendelian randomization(BWMR),and constrained maximum likelihood(cML)for validation.Multiple sensitivity analyses were performed to assess the robustness of the results.Results:Four metabolites were identified to have significant causal associations with PC risk.Higher phospholipid-to-total lipid ratios in intermediate-density lipoproteins(IDL)(GCST90445881)and small high density lipoproteins(HDL)(GCST90446027),as well as higher free cholesterol-to-total lipid ratios in extremely large very-low-density lipoproteins(VLDL)(GCST90446151),were inversely associated with PC risk.Conversely,an elevated triglyceride-to-total lipid ratio in chylomicrons and extremely large VLDL(GCST90446157)was positively associated with increased PC risk.The findings were consistently supported by multiple sensitivity analyses.Conclusion:This study provides genetic evidence linking lipid metabolism alterations to PC risk.Elevated phospholipid and free cholesterol ratios appear protective,whereas increased triglyceride levels act as risk factors.These metabolite profiles may serve as promising biomarkers for early diagnosis and intervention in PC,offering novel insights for risk assessment and potential metabolic-targeted therapies.

AIM:To investigate autophagic status in ischemic stroke with Liver Yang Hyperactivity and the mechanism of Tianma Gouteng Decoction(TMGTD).METHODS:SD rats were divided into sham,model,TMGTD high/medium/low-dose(20.52/10.26/5.13 g·kg-1·d-1),and Nimodipine(30 mg·kg-1·d-1)groups.A Liver Yang Hyperactivity and cerebral ischemia-reperfusion model was es-tablished using Fuzi Decoction(2 g·kg-1·d-1)and thread-occlusion.After 21 days of Fuzi decoction pretreatment,rats received daily drug administra-tion for 12 days.Syndrome indicators(irritability,24-hour water intake,24-hour urine volume,facial temperature)were recorded,plasma NE,E,cAMP,and cGMP were measured by ELISA,neurological function was assessed using Zea Longa and mNSS methods,brain histopathology was evaluated by HE staining,protein expression of soluble/insoluble p62 and LC3B was detected by Western blot,au-tophagy-related genes were analyzed by PCR array,additionally,mRNA and protein levels of CXCR4 and CXCL12 were measured by qRT-PCR and Western blot.RESULTS:Compared to the sham group,the model group showed increased irritability,24-hours water intake,24-hours urine volume,facial temper-ature,and level of NE,E,cGMP(P＜0.01),neurologi-cal scores(P＜0.01),LC3B-Ⅱ,insoluble p62,CXCR4,CXCL12 expression(P＜0.01),but decreased soluble p62(P＜0.01).TMGTD groups exhibited reduced irri-tability,water intake,urine volume,facial tempera-ture,NE,E,cGMP(P＜0.05,P＜0.01),neurological scores(P＜0.05,P＜0.01),p62 expression(P＜0.01),alongside increased LC3B-Ⅱ(P＜0.01)and improved cortical pathology.TMGD also reversed dysregulat-ed autophagy-apoptosis genes(CXCR4,Lamp1,Tgfb1,APP,Rab24)and reduced CXCR4,CXCL12 ex-pression(P＜0.01).CONCLUSION:In the Liver Yang Hyperactivity and cerebral ischemia-reperfusion model,autophagy genes were activated but flux was impaired,and Tianma Gouteng Decoction may protect by restoring autophagic flux and inhibiting the CXCL12/CXCR4 axis.

Background and Aims:Pancreatic cancer(PC)is a highly lethal gastrointestinal malignancy with poorly understood pathogenesis.Previous studies suggest that alterations in plasma metabolomics may be associated with PC development;however,traditional observational studies are prone to confounding and reverse causation,making it difficult to establish causal relationships.This study employed a two-sample Mendelian randomization(MR)approach to systematically evaluate the potential causal relationship between 325 nuclear magnetic resonance(NMR)metabolites and PC risk.Methods:Genome-wide association study(GWAS)data of 325 NMR metabolites from the UK Biobank were integrated with GWAS data of PC from FinnGen.Single nucleotide polymorphisms(SNPs)significantly associated with metabolites were selected as instrumental variables.The inverse variance weighted method served as the primary analysis,supplemented by MR-Egger regression,weighted median,weighted mode,Bayesian weighted Mendelian randomization(BWMR),and constrained maximum likelihood(cML)for validation.Multiple sensitivity analyses were performed to assess the robustness of the results.Results:Four metabolites were identified to have significant causal associations with PC risk.Higher phospholipid-to-total lipid ratios in intermediate-density lipoproteins(IDL)(GCST90445881)and small high density lipoproteins(HDL)(GCST90446027),as well as higher free cholesterol-to-total lipid ratios in extremely large very-low-density lipoproteins(VLDL)(GCST90446151),were inversely associated with PC risk.Conversely,an elevated triglyceride-to-total lipid ratio in chylomicrons and extremely large VLDL(GCST90446157)was positively associated with increased PC risk.The findings were consistently supported by multiple sensitivity analyses.Conclusion:This study provides genetic evidence linking lipid metabolism alterations to PC risk.Elevated phospholipid and free cholesterol ratios appear protective,whereas increased triglyceride levels act as risk factors.These metabolite profiles may serve as promising biomarkers for early diagnosis and intervention in PC,offering novel insights for risk assessment and potential metabolic-targeted therapies.

AIM:To investigate the impact of recombinant human CC16 protein(rhCC16)on cigarette smoke extract(CSE)-induced senescence-associated secretory phenotype(SASP)in human bronchial epithelial cells(HBECs)and in the lung tissues of chronic obstructive pulmonary disease(COPD)mice,and to explore the underlying mechanism.METHODS:HBECs were induced into cellular senescence using 5%CSE.The senescent HBECs were treated with 250 ng/mL rhCC16,and the levels of reactive oxygen species(ROS)were assessed using the 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)method.The levels of trimethylated histone H3 at lysine 9(H3K9me3),a marker of senescence-associated heterochromatic foci(SAHF),were detected using a Western blot assay.RT-qPCR and ELISA were utilized to measure the mRNA expression and protein levels of SASP components including interleukin-1 beta(IL-1β),IL-6,IL-8,chemokine(C-X-C motif)ligand-1(CXCL-1),matrix metalloproteinase 1(MMP1)and MMP3.Passive smoking was con-ducted for six months to induce COPD in mice.RhCC16(2.5 μg/g body weight)or an equal volume of PBS(20 μL)was intranasally administered from the 16th week of smoking in the COPD+rhCC16 group or COPD+PBS group,respectively,with administration 2 hours before smoking.ROS levels in lung tissue cells were investigated using DCFH-DA staining.H3K9me3 levels in lung tissues were tested using Western blot assay.RT-qPCR and ELISA were performed to examine the mRNA expression and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3.RESULTS:DCFH-DA staining results showed that CSE stimulation increased ROS levels in HBECs,while rhCC16 treatment reduced them(P＜0.01).Western blot results indicated that CSE stimulation elevated H3K9me3 levels in HBECs,which were decreased with rhCC16 treatment(P＜0.01).RT-qPCR and ELISA assays demonstrated that CSE stimulation upregulated the mRNA and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3 in HBECs,which were reduced with rhCC16 admin-istration(P＜0.05).DCFH-DA staining results showed an increase in ROS levels in the lung tissues of COPD mice,which were decreased with rhCC16 administration(P＜0.01).Western blot data revealed an increase in H3K9me3 levels in the lung tissues of COPD mice,which were reduced with rhCC16 treatment(P＜0.01).RT-qPCR and ELISA assays demon-strated an upregulation of the mRNA and protein levels of IL-1β,IL-6,IL-8,CXCL-1,MMP1 and MMP3 in the lung tis-sues of COPD mice,which were reduced with rhCC16 treatment(P＜0.05).No statistically significant differences were ob-served in the above-mentioned indicators between the lung tissues of COPD and COPD+PBS mice(P＞0.05).CONCLU-SION:rhCC16 can effectively inhibit CSE-induced SASP in HBECs and in the lung tissues of COPD mice,with its under-lying mechanism potentially related to the inhibition of the ROS-H3K9me3 signaling pathway.