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:We have successfully established an animal model of small ear pig in southern Yunnan province with internal tension relieving technique combined with autologous Achilles tendon for anterior cruciate ligament reconstruction,and verified the stability and reliability of the model.However,whether internal tension relieving technique can promote the ligamentalization process of autologous Achilles tendon graft has not been studied. OBJECTIVE:To investigate the differences in the process of ligamentalization between conventional reconstruction and internal reduction reconstruction of the anterior cruciate ligament by gross view,histology and electron microscopy. METHODS:Thirty adult female small ear pigs in southern Yunnan province were selected.Anterior cruciate ligament reconstruction was performed on the left knee joint with the ipsilateral knee Achilles tendon(n=30 in the normal group),and anterior cruciate ligament reconstruction was performed on the right knee joint with the ipsilateral knee Achilles tendon combined with the internal relaxation and enhancement system(n=30 in the relaxation group).The autogenous right forelimb was used as the control group;the anterior cruciate ligament was exposed but not severed or surgically treated.At 12,24,and 48 weeks after surgery,10 animals were sacrificed,respectively.The left and right knee joint specimens were taken for gross morphological observation to evaluate the graft morphology.MAS score was used to evaluate the excellent and good rate of the ligament at each time point.Hematoxylin-eosin staining was used to evaluate the degree of ligament graft vascularization.Collagen fibers and nuclear morphology were observed,and nuclear morphology was scored.Ultrastructural remodeling was evaluated by scanning electron microscopy and transmission electron microscopy. RESULTS AND CONCLUSION:(1)The ligament healing shape of the relaxation group was better at various time points after surgery,and the excellent and good rate of MAS score was higher(P<0.05).Moreover,the relaxation group could obtain higher ligament vascularization score(P<0.05).(2)The arrangement of collagen bundles and fiber bundles in the two groups gradually tended to be orderly,and the transverse fiber connections between collagen gradually increased and thickened,suggesting that the strength and shape degree of the grafts were gradually improved,but the ligament remodeling in the relaxation group was always faster than that in the normal group at various time points after surgery.(3)The diameter,distribution density,and arrangement degree of collagen fibers in the relaxation group were better than those in the normal group at all time points,especially in the comparison of collagen fiber diameter between and within the relaxation group(P<0.05).

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.

Background and Aims:Pancreatic cancer(PC)is a highly lethal malignancy with poor prognosis and limited early diagnostic tools.Although numerous serum metabolites have been associated with PC risk in observational studies,the causal nature of these associations remains uncertain.This study aimed to evaluate the genetic causal relationships between serum metabolites and PC risk,identify PC-related risk genes,and elucidate the gene-metabolite-PC causal network.Methods:Two-sample Mendelian randomization(TSMR)and summary-data-based Mendelian randomization(SMR)analyses were performed by integrating GWAS data of 325 serum metabolites,GTEx v8 pancreatic tissue eQTL data,and FinnGen R12 PC GWAS data.The study assessed causal effects of metabolites on PC risk,identified risk-associated genes,and explored the potential mediating role of metabolites between genes and PC.Results:Four serum metabolites showed significant causal relationships with PC risk.Elevated serum albumin(OR=1.456,P=0.003)and free cholesterol percentage in small high-density lipoprotein(HDL)(OR=1.189,P=0.005)were associated with increased PC risk,whereas higher phospholipid percentages in intermediate-density lipoprotein(IDL)and small HDL were protective(OR=0.792 and 0.836,respectively;both P<0.01).SMR analysis identified 196 PC-related genes,including risk genes such as DGKQ,CDC37P1,and SULT1A2,and protective genes such as PALMD and HEG1.Thirty-two significant gene-metabolite causal pairs were further confirmed,indicating potential mediation of PC genetic risk through specific metabolic pathways.Conclusion:This study systematically clarified the causal relationships between serum metabolites and pancreatic cancer risk and established a gene-metabolite regulatory network.The findings highlight the central role of lipid metabolism in PC development and provide molecular evidence for early detection and personalized prevention strategies.

Background and Aims:Pancreatic cancer(PC)is a highly lethal malignancy with poor prognosis and limited early diagnostic tools.Although numerous serum metabolites have been associated with PC risk in observational studies,the causal nature of these associations remains uncertain.This study aimed to evaluate the genetic causal relationships between serum metabolites and PC risk,identify PC-related risk genes,and elucidate the gene-metabolite-PC causal network.Methods:Two-sample Mendelian randomization(TSMR)and summary-data-based Mendelian randomization(SMR)analyses were performed by integrating GWAS data of 325 serum metabolites,GTEx v8 pancreatic tissue eQTL data,and FinnGen R12 PC GWAS data.The study assessed causal effects of metabolites on PC risk,identified risk-associated genes,and explored the potential mediating role of metabolites between genes and PC.Results:Four serum metabolites showed significant causal relationships with PC risk.Elevated serum albumin(OR=1.456,P=0.003)and free cholesterol percentage in small high-density lipoprotein(HDL)(OR=1.189,P=0.005)were associated with increased PC risk,whereas higher phospholipid percentages in intermediate-density lipoprotein(IDL)and small HDL were protective(OR=0.792 and 0.836,respectively;both P<0.01).SMR analysis identified 196 PC-related genes,including risk genes such as DGKQ,CDC37P1,and SULT1A2,and protective genes such as PALMD and HEG1.Thirty-two significant gene-metabolite causal pairs were further confirmed,indicating potential mediation of PC genetic risk through specific metabolic pathways.Conclusion:This study systematically clarified the causal relationships between serum metabolites and pancreatic cancer risk and established a gene-metabolite regulatory network.The findings highlight the central role of lipid metabolism in PC development and provide molecular evidence for early detection and personalized prevention strategies.

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.

OBJECTIVE To study the improvement effect of total flavonoids from Rosa multiflora root on vascular injury in rheumatoid arthritis （RA） model rats and its potential mechanism. METHODS Female Wistar rats were randomly divided into normal control group， model group， aspirin group （positive control， 30 mg/kg）， low-dose and high-dose groups of total flavonoids from R. multiflora root （4.15， 8.30 g/kg， by crude drug）， with 10 rats in each group. Except for the normal control group， the RA model was induced in other groups by collagen induction and high-fat diet. After 14 days of modeling， they were given corresponding drug solution/0.5% sodium carboxymethyl cellulose solution intragastrically， once a day， for 36 consecutive days. The total body score， arthritis index （AI） and swollen joint count （SJC） of the rats were evaluated regularly. After the last medication， serum levels of interleukin-6 （IL-6）， intercellular adhesion molecule-1 （ICAM-1） and vascular cell adhesion molecule- 1 （VCAM-1） were determined. The pathological morphological changes in the vascular tissue of thoracic aorta were observed； the protein expression of Toll-like receptor 4 （TLR4） and the protein phosphorylation levels of Janus kinase 2 （JAK2） and signal transduction and activator of transcription 3 （STAT3） in vascular tissue of thoracic aorta were measured. RESULTS Compared with the normal control group， serum levels of IL-6， ICAM-1 and VCAM-1， protein expression of TLR4， and the protein phosphorylation levels of JAK2 and STAT3 in vascular tissue of thoracic aorta were increased significantly in model group （P＜ 0.01）. The atherosclerotic plaque （atheroma）， cholesterol crystal， lymphocyte infiltration and a small number of unbroken foam cell aggregation could be seen in the vascular tissue of thoracic aorta. Compared with the model group， total body score （except for the low-dose group）， AI and SJC were decreased significantly in groups of total flavonoids from R. multiflora root on the 28th day （P＜0.05 or P＜0.01）； total body score，AI and SJC were decreased significantly in low-dose group of total flavonoids from R. multiflora root on the 49th day （P＜0.05 or P＜0.01）； the other quantitative indicators in serum and vascular tissue were significantly reversed in groups of total flavonoids from R. multiflora root （P＜0.05 or P＜0.01）， and pathological damage of vascular tissue was significantly relieved. CONCLUSIONS Total flavonoids from R. multiflora root can significantly improve vascular injury in RA model rats， and its mechanism may be related to reducing the protein expression of TLR4 in vascular tissue and inhibiting the activation of IL-6/JAK2/ STAT3 signaling pathway.

BACKGROUND:As a dominant breed pig in southwest China,the southern Yunnan small-ear pig has been widely used as an experimental animal in the basic research of other disciplines,but there are still no reports on its application in anterior cruciate ligament reconstruction. OBJECTIVE:To establish a large animal model of the southern Yunnan small-ear pig with anterior cruciate ligament with autologous Achilles tendon was established. METHODS:Twenty adult female Yunnan small-ear pigs were equally randomized into two groups.In the autologous Achilles tendon group,the right knee anterior cruciate ligament was reconstructed with autologous Achilles tendon as a graft,while in the sham-operated group,a similar operation was performed on the right knee without any treatment of the anterior cruciate ligament.General conditions of each pig were observed and recorded before and 12 months after surgery.Ligaments and grafts were taken for gross observation and MAS scoring.Hematoxylin-eosin staining was performed to observe morphological characteristics of ligaments.The staining and arrangement of type I and type Ⅲ collagen were evaluated by immunohistochemistry.Transmission electron microscopy was used to observe the type,size,diameter,ratio,and distribution of collagen fibers in ligaments. RESULTS AND CONCLUSION:All animals had normal diet and activity,good wound healing,no obvious inflammatory reaction,no local purulent infection,and no significant changes in mental and urinary conditions compared with those before surgery.The reconstructed cruciate ligament of the knee was intact,with no stiffness and normal range of motion.Both the anterior drawer and Lachman tests were negative.Gross observation of the graft:12 months after surgery,the grafts was in good position,with good integrity,obvious tension,ligament color close to the original anterior cruciate ligament,and complete surface synovial coverage.Most of the intraarticular ligaments in the autologous Achilles tendon group were defined as MAS I type and a few were defined as MAS Ⅱ type.In the sham-operated group,the intraarticular ligament was defined as MAS I type.Hematoxylin-eosin staining indicated that,12 months after surgery,collagen fibers in the autologous Achilles tendon group began to appear bundled,isotropic,and uniformly arranged,with more obvious isotropic corrugations,and the nuclei were mainly linear or spindle-shaped,which were similar to those in normal anterior cruciate ligament tissue of the sham-operated group.Immunohistochemistry results indicated that,12 months after surgery,there was a higher expression of type I collagen and significantly less expression of type Ⅲ collagen in the reconstructed anterior cruciate ligament in the autologous Achilles tendon group.The degree of type I and type Ⅲ staining was similar in the two groups.Under the transmission electron microscope,the diameter,arrangement and density of collagen fibers in the reconstructed anterior cruciate ligament of the autologous Achilles tendon group were similar to those of the original anterior cruciate ligament at 12 months after surgery,indicating that the ligament remodeling process had been basically completed in the autologous Achilles tendon group at 12 months after surgery.Through a comprehensive evaluation of animal general conditions,ligament general view,MAS score,hematoxylin-eosin staining,immunohistochemistry,and transmission electron microscopy observation,we successfully established a large animal model of anterior cruciate ligament reconstruction using autogenous Achilles tendon in southern Yunnan small-ear pigs,with good morphological,histological and ultrastructural results.