L-citrulline is a nonprotein amino acid that plays an important role in human health and has great market demand. Although microbial cell factories have been widely used for biosynthesis, there are still challenges such as genetic instability and low efficiency in the biosynthesis of L-citrulline. In this study, an efficient, plasmid-free, non-inducible L-citrulline-producing strain of Escherichia coli BL21(DE3) was engineered by combined strategies. Firstly, a chassis strain capable of synthesizing L-citrulline was constructed by block of L-citrulline degradation and removal of feedback inhibition, with the L-citrulline titer of 0.43 g/L. Secondly, a push-pull-restrain strategy was employed to enhance the L-citrulline biosynthesis, which realized the L-citrulline titer of 6.0 g/L. Thirdly, the NADPH synthesis and L-citrulline transport were strengthened to promote the synthesis efficiency, which achieved the L-citrulline titer of 11.6 g/L. Finally, fed-batch fermentation was performed with the engineered strain in a 3 L fermenter, in which the L-citrulline titer reached 44.9 g/L. This study lays the foundation for the industrial production of L-citrulline and provides insights for the modification of other amino acid metabolic networks.
Citrulline/biosynthesis* ; Escherichia coli/genetics* ; Metabolic Engineering/methods* ; Fermentation ; NADP/biosynthesis*

Objective: To evaluate the long-term effectiveness of an intelligent blood transfusion system in intraoperative blood management by comparing its performance with clinicians' decisions. Methods: A retrospective analysis of 26 760 surgical cases (2017-2024) was conducted, comparing pre- and post-implementation (2017-2019 vs 2020-2024) metrics, including transfusion prediction accuracy, rationality of blood use, and clinical outcomes. The system, powered by XGBoost, integrated patient demographics, laboratory results, and surgical data to predict red blood cell transfusion needs. Results: The intelligent blood transfusion systems achieved an accuracy of 80.62% in predicting transfusion necessity, significantly outperforming clinicians (24.83%, P<0.001). Its blood-use rationality rate was 83.92% vs 18.02% for clinicians (P<0.001). Post-implementation, major surgeries (grades Ⅲ-Ⅳ) increased while the requested blood units decreased. High physician compliance (>75%) correlated with 88.18% rationality. Conclusion: The intelligent blood transfusion system significantly improves the accuracy of transfusion decision-making, reduces excessive red blood cell use, optimizes perioperative transfusion management, and enhances the utilization of blood medical resources.

Objective To assess the correlation between the body mass index(BMI)and short-term prognosis in the patients with acute myocardial infarction complicating cardiogenic shock(AMI-CS).Methods A total of 262 patients with diagnosed AMI-CS hospitalized in this hospital from January 2013 to February 2022 were selected as the study subjects,and a total of 255 patients with AMI-CS were included after excluding the patients with incomplete clinical data,no coronary angiography,loss to follow-up and low BMI(<18.5 kg/m2).Among them,those with BMI of 18.5-<24.0 kg/m2 were included in the normal BMI group(n=150),and those with BMI of≥24.0 kg/m2 were included in the overweight or obese group(n=105).The baseline data,auxiliary examinations and treatment measures of the patients were collected,and the patients who did not die at the time of discharge were followed up by telephone on 30 d after AMI onset,and the survival status of the patients,whether having major adverse cardiovascular events(MACE)occurrence and the occurrence time were obtained.The correlation between BMI and endpoint events was assessed by the COX regression model.Results The proportion of complicating atrial fibrillation,BNP and urea levels in the overweight or obesity group were lower than those in the normal BMI group,the BMI and Hb levels were higher than those in the normal BMI group,and the differences were statistically significant(P<0.05).There was no statistically significant difference in the results of cardiovascular examination between the two groups(P>0.05).The use rate of β-receptor blockers in the overweight or obese group was higher than that in the normal BMI group,and the difference was statistically significant(P<0.05).On 30 d of follow-up,the all-cause mortality rate in the overweight or obese group was 39.0%,which was lower than 44.0%in the normal BMI group,but the difference was not statistically significant(P=0.430);there was no statistically signifi-cant difference in the incidence rate of each MACE event and the total incidence rate of MACE between the two groups(P>0.05).The BMI normal group served as the control,the risk of death and the occurrence risk of MACE events in the overweight or obese group had no significant increase(P>0.05).The short-term prognosis had no significant interaction between BMI and the patients in different subgroups(P>0.05).Con-clusion The short-term prognosis has no significant difference between the overweight/obese AMI-CS pa-tients and normal BMI AMI-Cs patients.

Objective To evaluate the effects of the method of blood-cooling and blood stasis-resolving on heart function and prognosis in patients with sepsis-induced myocardial dysfunction(SIMD).Methods Sixty patients with SIMD admitted to the department of critical care medicine of Affiliated Hospital of Nanjing University of Chinese Medicine from June 2022 to October 2024 were enrolled as study subjects.The patients were divided into treatment group and control group according to random number table,with 30 patients in each group.All patients received conventional treatments,the patients in the treatment group were given Taohe Chengqi decoction(Persicae Semen 12 g,Chinese rhubarb 12 g,Cinnamon twig 6 g,Licorice root 6 g and Sodium Sulfate 6 g),the decoction was concentrated to 200 mL and taken in 2 divided doses in the morning and evening,one dose daily;and the patients in the control group were given the same amount of warm water.The total course of treatment lasts for 7 days.The differences in indicators of cardiac function[brain natriuretic peptide(BNP),cardiac troponin I(cTnI),MB isoenzyme of creatine kinase(CK-MB),aspartate aminotransferase(AST)]and echocardiographic parameters[left ventricular ejection fraction(LVEF),left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD),mitral orifice early/late diastolic blood flow velocity ratio(E/A ratio),E/mitral and tricuspid valve ostia and the peak early diastolic velocity(E/e')]at admission,at 1st and 7th day after treatment,and prognosis indexes[mechanical ventilation time,intensive care unit(ICU)length of stay,total hospital stay,28-day survival rate]were compared between two groups.Draw the Kaplan-Meier survival curve and compare the difference in the 28-day cumulative survival rate between the two groups of patients.Results After 7 days therapy,LVEF of the treatment group was significantly higher than that the control(0.524±0.132 vs.0.458±0.118,P<0.05)and E/e'ratio of the treatment group was significantly lower than the control group[11.17(9.57,12.04)vs.11.82(11.28,13.72),P<0.05].There were no significant differences in the 28-day mortality and total hospital stay time between the two groups,but mechanical ventilation time[days:7.00(0.00,11.00)vs.12.50(3.50,21.75),P<0.05]and stay time of ICU[days:14.50(7.75,25.00)vs.21.00(14.25,31.50),P<0.05]in the treatment group were shorter than those in the control group.The Kaplan-Meier survival showed that the cumulative 28-day survival rate was similar between two groups(Log-Rank:χ2=1.448,P=0.229).Conclusion The method of blood-cooling and blood stasis-resolving could decrease mechanical ventilation time and length of stay in ICU of SIMD patients and could increase LVEF in the treatment group.

A 13-year-old boy was admitted to the hospital due to 1 month after trauma in a traffic accident on September 29，2017. After admission，due to a large amount of pleural effusion through the chest tube，thoracoscopic exploration and fiberboard dissection were performed，and the postoperative pleural effusion was not significantly reduced. The creatinine of pleural effusion was 913 μmol/L，CT showed that left hydronephrosis and perirenal encapsulated effusion were closely related to the ureter，so ureteral injury and urinothorax were considered. Retrograde ureterography revealed a defect of left ureter，approximately 6 cm in length，and left nephrectomy + left kidney rupture repair + renal autotransplantation was performed. 7 years later，the patient recovered well and the renal blood supply was good，no left hydronephrosis was found by MRU. Early diagnosis of ureteral injury is challenging，especially when accompanied by urinothorax. When life is stable，early diagnosis and treatment can reduce the loss of renal function after combined trauma. Renal autotransplantation is safe and effective for the treatment of long segmental ureteral defects.

For centuries, people have been searching for ways to reconstruct the mutilated thumb and fingers. Among the hundreds of operation methods that have appeared, the method of toe transplantation to reconstruct the thumb and fingers, which appeared in the second half of the 19 th century, had the best effect. However, due to the limitation of technical level at that time, only the pedicled toe could be transplanted to reconstruct the thumb and fingers. During the treatment period, the patient was in an inappropriate position where the hand and foot were fixed together for a long time, and the nerve was not repaired, so the thumb and fingers reconstructed after surgery had poor feeling. Therefore, it has not been widely used. It was not until 1966 when Yang Dongyue succussed in reconstructing the thumb using a free toe transplant with blood vessel anastomosis that toe transplantation gradually became the mainstream method of thumb and finger reconstruction. The appearance and function of the thumb and finger reconstructed by toe transplantation are still very different from that of the normal thumb and finger. Moreover, when multiple thumbs and fingers are defective, the transplantation of multiple toes for repair will cause great damages to the foot, so it is not suitable to reconstruct more than three thumb and fingers using toes in the same period. In 2007, Wang Zengtao proposed the concept of "full-finger reconstruction of thumb and fingers" and a series of new operation methods: new fingers were designed and assembled by means of using a variety of tissue combination assembly, which changed the traditional method of toe transplantation to reconstruct thumb and fingers, and the method of replacing thumb and fingers by toes was changed to manufacturing new thumb and fingers so that the toes could be retained and the thumb and fingers could be reconstructed with approximately normal shape and function. In recent years, the concept and series of new operation methods of thumb and finger reconstruction have been popularized at home and abroad. This paper focuses on the development of full-finger reconstruction of thumb and fingers.

Vertebral refracture following percutaneous vertebral augmentation (PVA) is commonly seen in elderly patients with osteoporotic thoracolumbar compression fractures (OTLCF). It can lead to recurrent pain, loss of vertebral height, progression of kyphosis, and even neurological dysfunction, significantly impairing patients′ quality of life. Current diagnosis and treatment face multiple challenges, including high misdiagnosis rate, difficulty in choosing between surgical and non-surgical treatment options, lack of standardized surgical protocols, interference from intralesional bone cement during procedures, inadequate stability of internal fixation in osteoporotic bone, and suboptimal compliance of anti-osteoporotic therapy. Establishing a standardized diagnostic and therapeutic framework is urgently needed. To standardize the management process and improve outcomes for vertebral refractures after PVA in elderly OTLCF patients, Spinal Trauma Group of the Orthopedic Branch of Chinese Medical Doctor Association organized experts in the field to develop Guideline for the diagnosis and treatment of vertebral refracture after percutaneous vertebral augmentation in elderly patients with osteoporotic thoracolumbar compression fractures ( version 2025), based on current literature and clinical experience, and adhering to principles of scientific rigor and clinical applicability. A total of 11 recommendations were proposed, encompassing diagnosis, treatment, and rehabilitation of vertebral refracture after PVA in elderly patients with OTLCF, aiming to provide a foundation for a standardized management.

The catalytic activity of 3-mercaptopyruvate (3MP) sulfurtransferase (MPST) converts 3MP to hydrogen sulfide (H₂S). However, the regulatory mechanisms governing MPST and its impact on the brain remain largely unexplored. Our study reveals the neuroprotective role of endothelial MPST-generated H₂S, regulated by protein phosphatase 2A (PP2A). Bioinformatics analysis and RNA sequencing demonstrated that endothelial PP2A is associated with neurodegenerative disease pathways. Cerebral ischemic mice exhibited significant inactivation of endothelial PP2A, evidenced by the reduction of PP2Acα in the brain endothelium. Mice with endothelium-specific null PP2A (PP2A^EC-cKO) exhibited neuronal loss, cognitive dysfunction, and long-term potentiation deficits. Postnatal inactivation of endothelial PP2A also contributes to cognitive dysfunction and neuronal loss. However, regaining endothelial PP2A activity by overexpressing Ppp2ca rescued neuronal dysfunction. Mechanistically, PP2A deficiency is intricately linked to the MPST-H₂S signaling pathway. A robust reduction in endothelial MPST-dependent H₂S production followed PP2A deficiency. Exogenous H₂S treatment and AAV-mediated overexpression of MPST in brain endothelial cells significantly mitigated neuronal dysfunction in PP2A^EC-cKO mice. Furthermore, PP2A deficiency promotes an increase in calcium influx and calpain2 phosphorylation, subsequently leading to MPST degradation. The PP2A activator (FTY720) and MPST activator (3MP sodium) both remarkably restored endothelial MPST-dependent H₂S production, subsequently rescuing ischemia-induced neurological deficits. In conclusion, our study demonstrates that endothelial PP2A deficiency leads to MPST degradation by activating calpain2, thus damaging neuronal function.

Ischemic stroke is the leading cause of disability and mortality worldwide. The blood‒brain barrier (BBB) is the first line of defense after ischemic stroke. Disruption of the BBB induced by brain microvascular endothelial cells (BMECs) dysfunction is a key event that triggers secondary damage to the central nervous system, where blood-borne fluids and immune cells penetrate the brain parenchyma, causing cerebral edema and inflammatory response and further aggravating brain damage. Here, we develop a novel artificial mesenchymal stem cell (MSC) extracellular vesicles by integrating MSC membrane proteins into liposomal bilayers, which encapsulated miR-132-3p with protective effects on BMECs. The artificial extracellular vesicles (MSCo/miR-132-3p) had low immunogenicity to reduce non-specific clearance by the mononuclear phagocytosis system (MPS) and could target ischemia-injured BMECs. After internalization into the damaged BMECs, MSCo/miR-132-3p escaped the lysosomes via the H_II phase transition of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) and decreased cellular reactive oxygen species (ROS) and apoptosis levels by regulating the RASA1/RAS/PI3K/AKT signaling pathway. In the transient middle cerebral artery occlusion (tMCAO) models, MSCo/miR-132-3p targeted impaired brain regions (approximately 9 times the accumulation of plain liposomes at 12 h), reduced cerebral vascular disruption, protected BBB integrity, and decreased infarct volume (from 44.95% to 6.99%).

OBJECTIVE: To assess the impact of albumin (Alb) administration on the prognosis of patients with acute kidney injury (AKI). METHODS: Clinical data of AKI patients in the intensive care unit (ICU) were retrospectively analyzed from the American Medical Information Mart of Intensive Care-IV (MIMIC-IV), including demographic data, acute physiology score (APS), comorbidities, vital signs, laboratory indicators, treatment status, ICU length of stay, and outcome indicators. The main outcome measure is ICU mortality. AKI patients were divided into Alb infusion group and Alb non infusion group based on whether they received Alb treatment. Multiple imputation was used to process missing data and eliminate variables that missing more than 30%. To ensure the stability of the results, propensity score matching (PSM) and inverse probability weighting (IPW) were used to correct the results. Using Kaplan-Meier survival curve and Cox proportional hazards regression model to evaluate the effect of Alb infusion on ICU survival rate in AKI patients. Perform subgroup analysis based on patient age, gender, and comorbidities to evaluate the prognostic effects of Alb on different patient subgroups. RESULTS: A total of 6 390 AKI patients were included, including 1 721 in the Alb infusion group and 4 669 in the Alb non infusion group. After adjusting for key covariates in the Cox regression model, compared with the Alb non infusion group, patients in the Alb infusion group were significantly younger in age, with APS III score, proportion of vasoactive drugs and continuous renal replacement therapy (CRRT) use, sepsis proportion, heart rate, respiratory frequency, aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine (Cr), lactic acid (Lac), and arterial partial pressure of carbon dioxide (PaCO₂) levels significantly higher. The proportion of hypertension, myocardial infarction, and congestive heart failure, as well as blood pressure, urine output, platelet count (PLT), and Alb levels were significantly lower. The results of univariate and multivariate Cox regression analysis on the raw data showed that the risk of death in the Alb infusion group was significantly lower than that in the Alb non infusion group [hazard ratio (HR) = 0.69, 95% confidence interval (95%CI) was 0.60-0.80, all P < 0.05]. The results after propensity score matching (PSM) and inverse probability weighting (IPW) processing are consistent with the original data trend (both P < 0.05). The Kaplan-Meier survival curve showed that the cumulative survival rate during ICU stay in the Alb infusion group was significantly higher than that in the Alb non infusion group (24.48% vs. 12.17%, Log-Rank test: χ² = 74.26, P < 0.05). Subgroup analysis shows that Alb infusion has a more significant survival benefit for AKI patients who use vasoactive drugs, have concurrent sepsis, and do not have liver disease. CONCLUSION Albumin infusion can decrease the ICU mortality of AKI patients.
Humans ; Retrospective Studies ; Acute Kidney Injury/mortality* ; Prognosis ; Male ; Female ; Middle Aged ; Aged ; Intensive Care Units ; Albumins/therapeutic use* ; Proportional Hazards Models ; Adult ; Databases, Factual