Metabolomics covers a wide range of applications in life sciences, biomedicine, and phytology. Data acquisition (to achieve high coverage and efficiency) and analysis (to pursue good classification) are two key segments involved in metabolomics workflows. Various chemometric approaches utilizing either pattern recognition or machine learning have been employed to separate different groups. However, insufficient feature extraction, inappropriate feature selection, overfitting, or underfitting lead to an insufficient capacity to discriminate plants that are often easily confused. Using two ginseng varieties, namely Panax japonicus (PJ) and Panax japonicus var. major (PJvm), containing the similar ginsenosides, we integrated pseudo-targeted metabolomics and deep neural network (DNN) modeling to achieve accurate species differentiation. A pseudo-targeted metabolomics approach was optimized through data acquisition mode, ion pairs generation, comparison between multiple reaction monitoring (MRM) and scheduled MRM (sMRM), and chromatographic elution gradient. In total, 1980 ion pairs were monitored within 23 min, allowing for the most comprehensive ginseng metabolome analysis. The established DNN model demonstrated excellent classification performance (in terms of accuracy, precision, recall, F1 score, area under the curve, and receiver operating characteristic (ROC)) using the entire metabolome data and feature-selection dataset, exhibiting superior advantages over random forest (RF), support vector machine (SVM), extreme gradient boosting (XGBoost), and multilayer perceptron (MLP). Moreover, DNNs were advantageous for automated feature learning, nonlinear modeling, adaptability, and generalization. This study confirmed practicality of the established strategy for efficient metabolomics data analysis and reliable classification performance even when using small-volume samples. This established approach holds promise for plant metabolomics and is not limited to ginseng.

Metabolomics covers a wide range of applications in life sciences,biomedicine,and phytology.Data acquisition(to achieve high coverage and efficiency)and analysis(to pursue good classification)are two key segments involved in metabolomics workflows.Various chemometric approaches utilizing either pattern recognition or machine learning have been employed to separate different groups.However,insufficient feature extraction,inappropriate feature selection,overfitting,or underfitting lead to an insufficient capacity to discriminate plants that are often easily confused.Using two ginseng varieties,namely Panax japonicus(PJ)and Panax japonicus var.major(PJvm),containing the similar ginsenosides,we integrated pseudo-targeted metabolomics and deep neural network(DNN)modeling to achieve accurate species differentiation.A pseudo-targeted metabolomics approach was optimized through data acquisition mode,ion pairs generation,comparison between multiple reaction monitoring(MRM)and scheduled MRM(sMRM),and chromatographic elution gradient.In total,1980 ion pairs were monitored within 23 min,allowing for the most comprehensive ginseng metabolome analysis.The established DNN model demonstrated excellent classification performance(in terms of accuracy,precision,recall,F1 score,area under the curve,and receiver operating characteristic(ROC))using the entire metabolome data and feature-selection dataset,exhibiting superior advantages over random forest(RF),support vector ma-chine(SVM),extreme gradient boosting(XGBoost),and multilayer perceptron(MLP).Moreover,DNNs were advantageous for automated feature learning,nonlinear modeling,adaptability,and generalization.This study confirmed practicality of the established strategy for efficient metabolomics data analysis and reliable classification performance even when using small-volume samples.This established approach holds promise for plant metabolomics and is not limited to ginseng.

BACKGROUND: The primary limitation to kidney transplantation is organ shortage. Recent progress in gene editing and immunosuppressive regimens has made xenotransplantation with porcine organs a possibility. However, evidence in pig-to-human xenotransplantation remains scarce, and antibody-mediated rejection (AMR) is a major obstacle to clinical applications of xenotransplantation. METHODS: We conducted a kidney xenotransplantation in a brain-dead human recipient using a porcine kidney with five gene edits (5GE) on March 25, 2024 at Xijing Hospital, China. Clinical-grade immunosuppressive regimens were employed, and the observation period lasted 22 days. We collected and analyzed the xenograft function, ultrasound findings, sequential protocol biopsies, and immune surveillance of the recipient during the observation. RESULTS: The combination of 5GE in the porcine kidney and clinical-grade immunosuppressive regimens prevented hyperacute rejection. The xenograft kidney underwent delayed graft function in the first week, but urine output increased later and the single xenograft kidney maintained electrolyte and pH homeostasis from postoperative day (POD) 12 to 19. We observed AMR at 24 h post-transplantation, due to the presence of pre-existing anti-porcine antibodies and cytotoxicity before transplantation; this AMR persisted throughout the observation period. Plasma exchange and intravenous immunoglobulin treatment mitigated the AMR. We observed activation of latent porcine cytomegalovirus toward the end of the study, which might have contributed to coagulation disorder in the recipient. CONCLUSIONS 5GE and clinical-grade immunosuppressive regimens were sufficient to prevent hyperacute rejection during pig-to-human kidney xenotransplantation. Pre-existing anti-porcine antibodies predisposed the xenograft to AMR. Plasma exchange and intravenous immunoglobulin were safe and effective in the treatment of AMR after kidney xenotransplantation.
Transplantation, Heterologous/methods* ; Kidney Transplantation/methods* ; Heterografts/pathology* ; Immunoglobulins, Intravenous/administration & dosage* ; Graft Survival/immunology* ; Humans ; Animals ; Sus scrofa ; Graft Rejection/prevention & control* ; Kidney/pathology* ; Gene Editing ; Species Specificity ; Immunosuppression Therapy/methods* ; Plasma Exchange ; Brain Death ; Biopsy ; Male ; Aged

Milk-derived exosomes are nanoscale extracellular vesicles naturally present in mammalian milk and are rich in proteins,lipids,nucleic acids,and bioactive metabolites.They exhibit biological activities such as anti-inflammatory,antioxidant,and immunomodulatory effects.Owing to their unique advantages of low immunogenicity,excellent biocompatibility,stability,and natural targeting ability,milk-derived exosomes have emerged as promising natural nanocarriers,garnering significant attention in disease treatment and nutritional interventions.Sarcopenia,characterized by reduced skeletal muscle mass and diminished strength,is closely associated with aging,chronic inflammation,and oxidative stress.Recent studies have highlighted the potential role of milk-derived exosomes in combating sarcopenia.These exosomes can increase myotube diameter,enhance muscle mass and fiber cross-sectional area,and improve exercise performance metrics such as grip strength.Potential mechanisms may include promoting muscle anabolism,improving mitochondrial function through antioxidant mechanisms,and modulating the inflammatory microenvironments.However,challenges remain in their application for sarcopenia at present,such as the unclear mechanisms of key bioactive components[e.g.,microRNAs(miRNAs),L-ornithine,and milk fat globule-epidermal growth factor 8(MFG-E8)],and the targeting and stability of delivery systems need to be optimized.Future research should focus on the functional analysis of components in exosomes,optimization of delivery systems,and preclinical validation to promote their practical application in managing age-related muscle health.

Objective To observe the effect of a Jianpi Huayu Jiedu formula on the NLRP3-mediated pyroptosis pathway in gastric precancerous lesion(GPL)associated with Helicobacter pylori(Hp)infection.Methods A GPL mouse model was prepared using Hp suspension gavage combined with Atp4a gene-deficient mice.The Jianpi Huayu Jiedu formula was administered as an intervention.Gastric mucosal tissue pathological damage was observed using hematoxylin and eosin(HE)staining.The presence of intestinal metaplasia(IM)was assessed using Alcian Blue-Periodic Acid Schiff(AB-PAS)staining.Ultrastructural changes in cell organelles were observed using transmission electron microscopy.Enzyme-linked immunosorbent assay(ELISA)was used to measure levels of gastrin-17(G-17),pepsinogen I(PGI),and proinflammatory cytokines IL-1β and IL-18.The expression of molecules related to the pyroptosis pathway was detected using Western blot and real-time quantitative PCR(RT-qPCR).Results Compared to the control group,Hp-related GPL mice exhibited gastric mucosal atrophy accompanied by IM and dysplasia.Damage to mitochondria and endoplasmic reticulum in parietal cells was observed.Levels of G-17,PGI,and proinflammatory cytokines IL-1β and IL-18 were elevated.The expression of molecules related to the pyroptosis pathway was increased.The Jianpi Huayu Jiedu formula significantly reduced gastric mucosal tissue pathological damage in GPL mice,decreased G-17 and PGI levels,mitigated inflammatory responses,and downregulated the expression of molecules related to the pyroptosis pathway.Conclusion The Jianpi Huayu Jiedu formula may exert its effects by inhibiting the NLRP3-mediated pyroptosis signaling pathway,thereby alleviating or even reversing the pathological damage of gastric mucosa in Hp-related GPL.

Objective To observe the effect of a Jianpi Huayu Jiedu formula on the NLRP3-mediated pyroptosis pathway in gastric precancerous lesion(GPL)associated with Helicobacter pylori(Hp)infection.Methods A GPL mouse model was prepared using Hp suspension gavage combined with Atp4a gene-deficient mice.The Jianpi Huayu Jiedu formula was administered as an intervention.Gastric mucosal tissue pathological damage was observed using hematoxylin and eosin(HE)staining.The presence of intestinal metaplasia(IM)was assessed using Alcian Blue-Periodic Acid Schiff(AB-PAS)staining.Ultrastructural changes in cell organelles were observed using transmission electron microscopy.Enzyme-linked immunosorbent assay(ELISA)was used to measure levels of gastrin-17(G-17),pepsinogen I(PGI),and proinflammatory cytokines IL-1β and IL-18.The expression of molecules related to the pyroptosis pathway was detected using Western blot and real-time quantitative PCR(RT-qPCR).Results Compared to the control group,Hp-related GPL mice exhibited gastric mucosal atrophy accompanied by IM and dysplasia.Damage to mitochondria and endoplasmic reticulum in parietal cells was observed.Levels of G-17,PGI,and proinflammatory cytokines IL-1β and IL-18 were elevated.The expression of molecules related to the pyroptosis pathway was increased.The Jianpi Huayu Jiedu formula significantly reduced gastric mucosal tissue pathological damage in GPL mice,decreased G-17 and PGI levels,mitigated inflammatory responses,and downregulated the expression of molecules related to the pyroptosis pathway.Conclusion The Jianpi Huayu Jiedu formula may exert its effects by inhibiting the NLRP3-mediated pyroptosis signaling pathway,thereby alleviating or even reversing the pathological damage of gastric mucosa in Hp-related GPL.

Milk-derived exosomes are nanoscale extracellular vesicles naturally present in mammalian milk and are rich in proteins,lipids,nucleic acids,and bioactive metabolites.They exhibit biological activities such as anti-inflammatory,antioxidant,and immunomodulatory effects.Owing to their unique advantages of low immunogenicity,excellent biocompatibility,stability,and natural targeting ability,milk-derived exosomes have emerged as promising natural nanocarriers,garnering significant attention in disease treatment and nutritional interventions.Sarcopenia,characterized by reduced skeletal muscle mass and diminished strength,is closely associated with aging,chronic inflammation,and oxidative stress.Recent studies have highlighted the potential role of milk-derived exosomes in combating sarcopenia.These exosomes can increase myotube diameter,enhance muscle mass and fiber cross-sectional area,and improve exercise performance metrics such as grip strength.Potential mechanisms may include promoting muscle anabolism,improving mitochondrial function through antioxidant mechanisms,and modulating the inflammatory microenvironments.However,challenges remain in their application for sarcopenia at present,such as the unclear mechanisms of key bioactive components[e.g.,microRNAs(miRNAs),L-ornithine,and milk fat globule-epidermal growth factor 8(MFG-E8)],and the targeting and stability of delivery systems need to be optimized.Future research should focus on the functional analysis of components in exosomes,optimization of delivery systems,and preclinical validation to promote their practical application in managing age-related muscle health.

Identifying the compound formulae-related xenobiotics in bio-samples is full of challenges.Conventional strategies always exhibit the insufficiencies in overall coverage,analytical efficiency,and degree of automation,and the results highly rely on the personal knowledge and experience.The goal of this work was to establish a software-aided approach,by integrating ultra-high performance liquid chromatography/ion-mobility quadrupole time-of-flight mass spectrometry(UHPLC/IM-QTOF-MS)and in-house high-definition MS2 library,to enhance the identification of prototypes and metabolites of the compound formulae in vivo,taking Sishen formula(SSF)as a template.Seven different MS2 acquisition methods were compared,which demonstrated the potency of a hybrid scan approach(namely high-definition data-independent/data-dependent acquisition(HDDIDDA))in the identification precision,MS1 coverage,and MS2 spectra quality.The HDDIDDA data for 55 reference compounds,four component drugs,and SSF,together with the rat bio-samples(e.g.,plasma,urine,feces,liver,and kidney),were acquired.Based on the UNIFI? platform(Waters),the efficient data processing workflows were estab-lished by combining mass defect filtering(MDF)-induced classification,diagnostic product ions(DPIs),and neutral loss filtering(NLF)-dominated structural confirmation.The high-definition MS2 spectral li-braries,dubbed in vitro-SSF and in vivo-SSF,were elaborated,enabling the efficient and automatic identification of SSF-associated xenobiotics in diverse rat bio-samples.Consequently,118 prototypes and 206 metabolites of SSF were identified,with the identification rate reaching 80.51％and 79.61％,respectively.The metabolic pathways mainly involved the oxidation,reduction,hydrolysis,sulfation,methylation,demethylation,acetylation,glucuronidation,and the combined reactions.Conclusively,the proposed strategy can drive the identification of compound formulae-related xenobiotics in vivo in an intelligent manner.

Objective:To evaluate the effect of berberine on acute kidney injury (AKI) in rats undergoing liver transplantation and the role of AMP-activated protein kinase (AMPK).Methods:Twenty-four SPF-grade adult male Sprague-Dawley rats, aged 12 weeks, weighing 210-230 g, were divided into 4 groups ( n=6 each) using the random number table method: sham operation group (S group), AKI group, berberine group (BBR group), and berberine + AMPK inhibitor Compound C group (BBR-Comp C group). In BBR group, berberine 200 mg/kg was given by gavage starting from 2 weeks before surgery, once a day for 14 consecutive days. In BBR-Comp C group, Compound C 1 mg/kg was injected into the tail vein at 30 min before surgery. The rat AKI model was prepared by in situ liver transplantation in AKI group, BBR group and BBR-Comp C group. Blood specimens were taken from the inferior vena cava at 24 h postoperatively, and serum BUN and Cr concentrations were determined by enzyme-linked immunosorbent assay. Then the rats were sacrificed, and the kidney tissues were taken for microscopic examination of the pathological changes (with the light microscope after HE staining) and for determination of the expression of phosphorylated AMPK (p-AMPK), receptor-interacting protein kinase-1 (RIPK-1), receptor-interacting protein kinase-3 (RIPK-3) and mixed lineage kinase domain-like protein (MLKL) (by Western blot). Results:Compared with S group, the serum BUN and Cr concentrations were significantly increased, the p-AMPK expression was down-regulated, the expression of RIPK-1, RIPK-3 and MLKL was up-regulated ( P<0.05), and the pathological damage to renal tissues occurred in AKI group. Compared with AKI group, the serum BUN and Cr concentrations were significantly decreased, the p-AMPK expression was up-regulated, the expression of RIPK-1, RIPK-3 and MLKL was down-regulated ( P<0.05), and the pathological changes of renal tissues were significantly attenuated in BBR group. Compared with BBR group, the serum BUN and Cr concentrations were significantly increased, the p-AMPK expression was down-regulated, and the expression of RIPK-1, RIPK-3 and MLKL was up-regulated in BBR-Comp C group ( P<0.05). Conclusions:Berberine can attenuate AKI in rats undergoing liver transplantation, and the mechanism may be related to the promotion of AMPK phosphorylation and inhibition of programmed necrosis.

With the vigorous promotion of organ donation after citizen death in China, increased utilization of marginal livers, and continuous expansion of hepatocellular carcinoma indications for liver transplantation, innovations in techniques such as auxiliary liver transplantation, pediatric liver transplantation, laparoscopic liver transplantation, magnetic liver transplantation,and non-ischemic liver transplantation have significantly improved the number of liver transplantation surgery performed, patient survival rates, and graft survival rates in China, while complication rates have gradually decreased. As such,liver transplantation in China has reached leading or advanced levels internationally. In the new development context,Chinese liver transplantation faces new opportunities and challenges for development. Evolutions in basic diseases of transplant recipients and tumor classifications of will further broaden the population eligible for transplantation and introduce new demands for liver transplantation procedures. Emerging technologies including artificial organs, xenotransplantation,and artificial intelligence are bringing prospects for advancing liver transplantation. Looking ahead, the progression of liver transplantation will go beyond prioritizing patient survival rates and graft survival rates alone, instead emphasizing improved quality of life for transplant recipients post-surgery to an even greater extent.