Objective To establish an "human serum - porcine aortic endothelial cells (PAEC) - human platelets" in vitro model and explore the mechanism of xenotransplantation-related coagulation dysfunction mediated by immune complexes - platelet FcγRⅡa (CD32a) receptor. Methods Healthy human serum was co-incubated with PAEC to prepare the supernatant containing immune complexes, which was then used to stimulate healthy human platelets, or directly treated with the serum of xenogeneic liver transplant recipients. Flow cytometry was used to detect platelet activation markers CD62P and surface IgG binding levels, and the platelet adhesion function was evaluated by platelet-PAEC adhesion experiments. CD32a blocking antibody IV.3 and SYK blocker SKYIN 4 were used to clarify the signaling pathways. Results The supernatant from the co-incubation of healthy human serum and PAEC could significantly induce platelet activation and endothelial adhesion. The use of the serum from xenogeneic liver transplant recipients could also significantly induce platelet activation. Antibody IV.3 and SYK blocker SKYIN 4 could significantly inhibit these effects. Conclusions In xenotransplantation, the immune complexes formed by human serum antibodies and porcine endothelial antigens may induce abnormal platelet activation through the platelet CD32a receptor, which is an important mechanism of non-complement-dependent post-transplant coagulation dysfunction, providing a new target for the intervention of coagulation complications in xenotransplantation.

Xenotransplantation using pigs as donors is considered one of the most promising approaches to address the shortage of allogeneic organs and has been seen rapid development in recent years. However, the clinical application of xenotransplantation still faces challenges in immune rejection, physiological compatibility, and biosafety. Among these, humoral rejection is a major barrier affecting the long-term survival of xenografts, and human recipients may have pre-existing antibodies against triple gene knockout (TKO) donor pigs. Nevertheless, early antibody-mediated humoral rejection can still be prevented. Additionally, cellular rejection and inflammatory responses are closely related to the long-term survival of grafts, and the introduction of humanized molecules in donor pigs, combined with the use of anti-inflammatory drugs by recipients, can help improve the survival of xenografts. Moreover, physiological challenges are mainly reflected in the incompatibility of the coagulation cascade and the structural and functional differences between pig proteins and human proteins. In terms of biosafety, breeding donor pigs free of designed pathogens and developing highly sensitive detection methods for pig-derived microorganisms are key to reduce transplant rejection and interspecies infections. At present, further research is needed in gene editing strategies, immunosuppressive regimens, and infection monitoring methods to improve the long-term survival of grafts and recipients.

Liver transplantation is the only effective cure for end-stage liver disease. However,donor liver shortage has become a major problem,limiting the development of liver transplantation. Among the many ways to solve the problem of donor liver shortage,xenotransplantation has high feasibility and prospect of clinical application. In recent years,heart and kidney xenotransplantation have been successfully carried out in clinical trials,while there are few studies on liver xenotransplantation. The survival time of preclinical models in liver xenotransplantation is limited to 34 days,which is not sufficient for clinical trials. The future clinical trials of liver xenotransplantation will face multiple challenges such as immune rejection,interspecific incompatibility,social and ethical issues,which require the joint efforts of government,society,research institution and medical institution and to promote the basic and clinical research of liver xenotransplantation in China.

BACKGROUND:Nobiletin has been found to improve lipopolysaccharide-induced abnormal activation of microglia,excessive release of inflammatory factors and redox imbalance.However,the specific mechanism is not fully understood. OBJECTIVE:To investigate the molecular mechanism by which nobiletin can inhibit lipopolysaccharide-induced inflammation in BV2 microglia. METHODS:Passage 3 BV2 microglia were divided into three groups:control group was cultured for 24 hours(without any treatment).Lipopolysaccharide group was treated with 10 μg/mL lipopolysaccharide for 24 hours.Lipopolysaccharide+nobiletin group was treated with 20 μmol/L nobiletin for 6 hours and then 10 μg/mL lipopolysaccharide for 24 hours.After the processing,cell proliferation was detected by CCK-8 assay.The level of intracellular reactive oxygen species was detected by fluorescent probe.The mRNA expression levels of nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β were detected by qRT-PCR.The protein expression levels of nuclear factor κB p65,p-nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β were detected by western blot assay. RESULTS AND CONCLUSION:(1)Compared with the control group,the proliferation activity of lipopolysaccharide group was decreased(P<0.001).Compared with the lipopolysaccharide group,the cell proliferation activity of lipopolysaccharide+nobiletin group was increased(P<0.001).(2)Compared with the control group,the level of intracellular reactive oxygen species was increased in the lipopolysaccharide group(P<0.001).Compared with the lipopolysaccharide group,the level of intracellular reactive oxygen species was decreased in the lipopolysaccharide+nobiletin group(P<0.01).(3)Compared with the control group,the mRNA expression levels of tumor necrosis factor α and interleukin-1β were increased in the lipopolysaccharide group(P<0.001,P<0.01).Compared with the lipopolysaccharide group,mRNA expression levels of tumor necrosis factor α and interleukin-1β were decreased in the lipopolysaccharide+nobiletin group(P<0.01,P<0.05).(4)Compared with the control group,the protein expression levels of p-nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β in were increased the lipopolysaccharide group(P<0.001).Compared with the lipopolysaccharide group,the expression of p-nuclear factor κB p65,tumor necrosis factor α,and interleukin-1β was decreased in the lipopolysaccharide+nobiletin group(P<0.001).(5)These findings suggest that nobiletin attenuates lipopolysaccharide-induced inflammatory response in BV2 microglia by suppressing nuclear factor-κB signaling pathway.

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

Objective To investigate the mechanism of total ginsenosides(TG)in alleviating myocardial injury in septic rat models by regulating solute carrier family 7 member 11/glutathione per-oxidase 4(SLC7 A1 1/GPX4)-mediated ferroptosis.Methods Ninety SPF-grade male rats were ran-domly divided into five groups:Sham group(sham operation),septic cardiomyopathy(SCM)group,TG40 group(SCM model rats gavaged with 40 mg/kg TG),TG160 group(SCM model rats gavaged with 160 mg/kg TG),and TG160+RSL3 group(SCM model rats gavaged with 160 mg/kg TG and intraperitoneally injected with 10 mg/kg ferroptosis inducer RSL3).The SCM model was established using the cecal ligation and puncture method.Cardiac function was assessed in each group.Hematox-ylin-eosin(HE)staining was performed to observe pathological changes in myocardial tissue.Levels of tumor necrosis factor(TNF)-α,interleukin(IL)-1 β,IL-6,creatine kinase-MB(CK-MB),myo-globin(Mb),and cardiac troponin I(cTnI)were measured by enzyme-linked immunosorbent assay.Protein expression levels of NADPH oxidase(NOX)1,NOX2,NOX4,SLC7A11,and GPX4 were detected by western blotting.Levels of Fe2+and total iron in myocardial tissue were also determined.Results Compared with the Sham group,the SCM group exhibited decreased left ventricular ejec-tion fraction(LVEF),left ventricular fractional shortening(LVFS),and protein expression of SLC7A11 and GPX4,along with increased levels of CK-MB,Mb,cTnI,TNF-α,IL-1β,IL-6,Fe2+,total iron,and protein expression of NOX1,NOX2,and NOX4(P＜0.05).Compared with the SCM group,the TG40 and TG160 groups showed dose-dependent increases in LVEF,LVFS,and SLC7A11/GPX4 expression,as well as reductions in CK-MB,Mb,cTnI,TNF-α,IL-1 β,IL-6,Fe,total iron,and NOX1/NOX2/NOX4 expression(P＜0.05).In contrast,the TG160+RSL3 group displayed lower LVEF,LVFS,and SLC7A11/GPX4 expression,and higher levels of the aforementioned biomarkers compared with the TG160 group(P＜0.05).Conclusion TG sig-nificantly suppresses sepsis-induced myocardial inflammation and oxidative stress,improves cardiac function,and mitigates myocardial injury,likely by inhibiting ferroptosis via activation of the SLC7A11/GPX4 signaling pathway.

Nasojejunal feeding tubes are widely used in surgical,intensive care,and older patients.Manual blind insertion of nasojejunal feeding tubes is technically challenging,associated with a high failure rate,and prone to complications.The primary causes of suboptimal placement outcomes are the uncertainty and weak controllability of the interaction forces during the coordination between manual posterior advancement and the patient's physiological state.While current auxiliary techniques such as X-ray,ultrasound,and endoscopy can improve the success rate of nasojejunal tube placement and reduce complications to some extent,the accuracy and safety of placement remain constrained by challenges in controlling insertion forces and achieving precise positional localization.Robotic technology holds promise for addressing the uncertainties and controllability issues inherent in the placement process.By leveraging precise sensing,real-time navigation,and efficient control,robots can achieve intelligent positioning and precise control over the direction and location of the catheter tip during nasojejunal intubation.However,current research on robotic applications for nasojejunal feeding tube placement is still in an early stage,facing challenges such as high costs,operational complexity,and concerns over safety and reliability.Herein,we analyzed the limitations and causes of failure in existing placement methods and explored the application prospects of robotic technologies for precise control and intelligent positioning in nasojejunal feeding tube placement.The paper provides new insights for developing nursing techniques that enable safer and more effective,comfortable,and rapid intubation.Future efforts should focus on deepening the integration of artificial intelligence and robotics,optimizing drive technologies,and accelerating the translation of these technologies from the laboratory to clinical practice.This will drive the advancement of nasojejunal feeding tube placement techniques towards intelligent,precise,and accessible solutions.

Objective To achieve a preliminary understanding of the current situation of clinical laboratories in Guangdong Province,and discuss how to establish a sound investigation system,and utilize quality indicators to improve laboratory quality through the inves-tigation and analysis of data from 16 clinical laboratory quality indicators issued by the National Center for Clinical Laboratories.Meth-ods The questionnaire was issued by Clinet-EQA system and the basic information and quality indicator information during 2023 were collected.SPSS 20.0 software was used for statistical analysis according to different specialty categories and hospital grades.The 13 quality indicators measured in rate-based units were evaluated by sigma measurement.The P75,P50 and P25 percentiles of the overall distribution of each quality index were used to explore the optimal,appropriate and minimum quality specifications.Results A total of 577 laboratories participated in this survey.In addition to the implementation rate of internal quality assessment and the inter-laboratory comparison rate,the median sigma(σ)value of 11/13 quality indicators was greater than 3σ,and some of them even reach the level of 6σ,and there were disparities between hospital laboratories at different grades.The turnaround time(TAT)of the whole process of emergency examination was significantly less than those of inpatient and outpatient,TAT before emergency examination was controlled within 20 min,TAT before outpatient examination was within 30 min,and TAT before inpatient examination was within 42 min.The optimal quality specifications of 8 out of 13 indicators reached 6σ level,while the minimum quality specifications of 2 out of 13 indica-tors were lower than 3σ level.Conclusion In Guangdong Province,the overall level of quality indicators in the post-analytical of clin-ical laboratories was superior to that in the pre-analytical and analytical process.It should be essential to continuously monitor quality indicators and actively adopt improvement measures for those laboratories with unsatisfactory results,so as to enhance the examination quality of laboratories.

Objective To achieve a preliminary understanding of the current situation of clinical laboratories in Guangdong Province,and discuss how to establish a sound investigation system,and utilize quality indicators to improve laboratory quality through the inves-tigation and analysis of data from 16 clinical laboratory quality indicators issued by the National Center for Clinical Laboratories.Meth-ods The questionnaire was issued by Clinet-EQA system and the basic information and quality indicator information during 2023 were collected.SPSS 20.0 software was used for statistical analysis according to different specialty categories and hospital grades.The 13 quality indicators measured in rate-based units were evaluated by sigma measurement.The P75,P50 and P25 percentiles of the overall distribution of each quality index were used to explore the optimal,appropriate and minimum quality specifications.Results A total of 577 laboratories participated in this survey.In addition to the implementation rate of internal quality assessment and the inter-laboratory comparison rate,the median sigma(σ)value of 11/13 quality indicators was greater than 3σ,and some of them even reach the level of 6σ,and there were disparities between hospital laboratories at different grades.The turnaround time(TAT)of the whole process of emergency examination was significantly less than those of inpatient and outpatient,TAT before emergency examination was controlled within 20 min,TAT before outpatient examination was within 30 min,and TAT before inpatient examination was within 42 min.The optimal quality specifications of 8 out of 13 indicators reached 6σ level,while the minimum quality specifications of 2 out of 13 indica-tors were lower than 3σ level.Conclusion In Guangdong Province,the overall level of quality indicators in the post-analytical of clin-ical laboratories was superior to that in the pre-analytical and analytical process.It should be essential to continuously monitor quality indicators and actively adopt improvement measures for those laboratories with unsatisfactory results,so as to enhance the examination quality of laboratories.

Xenotransplantation using pigs as donors is considered one of the most promising approaches to address the shortage of allogeneic organs and has been seen rapid development in recent years. However, the clinical application of xenotransplantation still faces challenges in immune rejection, physiological compatibility, and biosafety. Among these, humoral rejection is a major barrier affecting the long-term survival of xenografts, and human recipients may have pre-existing antibodies against triple gene knockout (TKO) donor pigs. Nevertheless, early antibody-mediated humoral rejection can still be prevented. Additionally, cellular rejection and inflammatory responses are closely related to the long-term survival of grafts, and the introduction of humanized molecules in donor pigs, combined with the use of anti-inflammatory drugs by recipients, can help improve the survival of xenografts. Moreover, physiological challenges are mainly reflected in the incompatibility of the coagulation cascade and the structural and functional differences between pig proteins and human proteins. In terms of biosafety, breeding donor pigs free of designed pathogens and developing highly sensitive detection methods for pig-derived microorganisms are key to reduce transplant rejection and interspecies infections. At present, further research is needed in gene editing strategies, immunosuppressive regimens, and infection monitoring methods to improve the long-term survival of grafts and recipients.