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.

Cervical spinal cord injury without fracture-dislocation (CSCIWFD) is referred to as a special type of cervical spinal cord injury characterized by traumatic spinal cord dysfunction and no significant bony structural abnormalities on imagines. Duo to the high risk of missed diagnosis during the initial consultation, CSCIWFD may lead to progressive neurological deterioration or even complete paralysis, severely impacting patients′ prognosis. Currently, there are no established consensuses over the diagnosis and treatment of CSCIWFD, such as the lack of evidence-based standards for indications of non-surgical treatment and risk of secondary neurological injury, as well as debates over the optimal timing for surgical intervention and indications for different surgical approaches. To address these issues, the Spine Trauma Group of the Orthopedic Branch of the Chinese Medical Doctor Association organized experts in the relevant fields to formulate Diagnosis and treatment guideline for acute cervical spinal cord injury without fracture- dislocation in adults ( version 2025) . Based on evidence-based medicine and the principles of scientific rigor and clinical applicability, the guidelines proposed 11 recommendations covering terminology, diagnosis, evaluation treatment, and rehabilitation, etc., aiming to standardize the management of CSCIWFD.

Objective To investigate the organ protective role and the underlying mechanism of nafamostat mesylate(NM)in a renal ischemia-reperfusion injury(RIRI)model.Methods A total of 21 healthy male Sprague-Dawley(SD)rats were randomly assigned to 3 groups(n=7 in each group),including the sham operation group(Sham group),the RIRI group,and the NM intervention group(NM group).The RIRI and NM groups underwent ischemia-reperfusion injury(IRI)modeling.The NM group was given an intraperitoneal injection of NM at 0.75 mg/kg before modeling.Venous blood and renal tissue samples were then collected from the rats 24 hours after modeling.The levels of serum creatinine,cystatin C,and serum inflammatory factors were determined using the serum samples.Hematoxylin-eosin(HE)staining and TUNEL stainings were performed on the renal tissues to evaluate the damage of the renal tissues.The localization and expression of HMGB1 were analyzed by immunofluorescence and Western blotting,respectively.Single-cell RNA sequencing of the nuclei was performed to obtain the single-cell transcriptome of the kidneys from the rats in the RIRI and the NM groups and to acquire the RIRI cell profile.The cells were annotated according to the cell marker genes to explore the cell type composition in the disease model and the functional status of immune cells between the groups.Results 1)Compared with those of the Sham group,the levels of cystatin C,creatinine,and inflammatory factors in the RIRI and NM groups were significantly increased,and the expression levels in the NM group were lower than those in the RIRI group(P＜0.05).Compared with those of the RIRI group,the tubular injury score and apoptosis rate in the NM group were significantly decreased(P＜0.05),but those of both the NM and RIRI groups were higher than those of the sham group.Compared with that in the RIRI group,the expression of HMGB1 in the NM group was significantly decreased(P＜0.05),but the expression levels in both the RIRI and NM groups were higher than that in the sham group.Immunofluorescence showed that there was increased cytoplasmic expression of HMGB1 in both the NM and RIRI groups,with the increase being more prominent in the RIRI group.2)A total of 13 major cell populations were identified through the single-nucleus sequencing results.The proportion of tubular cells in the NM group was higher,with the HMGB1 gene being highly expressed in the damaged proximal convoluted tubular cells.The proportion of the polarized Macro3 cell subpopulation in the macrophages in the NM group was lower compared to that in the RIRI group.Conclusion NM may play a protective role in a rat model of RIRI,and its underlying mechanisms may be related to the regulation of the functional abnormalities of HMGB1-mediated macrophages.

Objective To explore the changes in neural activity in patients with type 2 diabetes mellitus(T2DM)and their corre-lation with executive function,and to analyze the neural mechanisms underlying the decline in executive function in T2DM patients.Methods Thirty-one T2DM patients(T2DM group)and thirty-two healthy controls(HC)(HC group)matched for body mass index(BMI)underwent resting-state functional magnetic resonance imaging(rs-fMRI)scans and N-back task tests were included.Differ-ences in the amplitude of low-frequency fluctuation(ALFF),regional homogeneity(ReHo),and seed-based functional connectivity(FC)between the two groups were compared,and partial correlation analyses were performed between the difference results and N-back task performance.Results The T2DM group showed prolonged reaction time(RT)in the 1-back and 2-back tasks.T2DM patients exhibited increased ALFF in the bilateral caudate nucleus,left medial superior frontal gyrus,and right postcentral gyrus,as well as elevated ReHo in the right putamen.FC analysis revealed significant alterations in FC between the caudate nucleus,putamen,and multiple brain regions in T2DM patients,with some of these FC changes significantly correlated with RT and accuracy(ACC)in the N-back task.Conclusion The decline in executive function in T2DM patients may be associated with abnormal neural activity in brain regions such as the striatum,salience network,and frontoparietal control network.FC further decreases under increased cognitive load.These findings provide evidence for the study of the neural mechanisms of executive function impairment in T2DM patients.

Objective To explore the clinical information of female patients with pulmonary nodules and the distribution of traditional Chinese medicine (TCM) syndromes and their elements. Methods A cross-sectional study was conducted to collect the basic information, medical history data, image data, and four diagnostic information of female patients with pulmonary nodules. The distribution characteristics of TCM syndromes and their elements in female patients with pulmonary nodules were determined by KMO test, Bartlett spherical test, systematic cluster analysis, chi-square test, and other methods. Results A total of 721 female patients with pulmonary nodules were included in this study. The patients were mainly 45-59 years old, had secondary school education or above, and had a history of oil smoke exposure as clinical characteristics. The pulmonary nodules were mainly 6-10 mm in size and appeared as multiple and ground glass nodules. The clinical symptoms were mainly fatigue, emotional irritability, and shortness of breath. The main syndromes of disease location were the spleen, liver, and lung; and the main syndromes of disease were phlegm, dampness, and qi deficiency. The main complex syndromes were spleen deficiency and dampness, liver stagnation, and qi/yin deficiency. Conclusion Middle age, high education, and multiple small ground glass nodules are the clinical characteristics of female patients with pulmonary nodules. Exposure to oil smoke is an important cause of the occurrence of female pulmonary nodules. During treatment, attention should be paid to strengthening the spleen, removing dampness, soothing the liver, regulating and tonifying qi, and nourishing yin.

Objective To investigate the initial temperature sensation and pressure pain sensation in patients receiving invisible or-thodontic treatment.Methods Twenty-two patients receiving clear aligner treatment(experimental group)and 22 volunteers(control group)participated.Perceptual changes were assessed using the visual analogue scale(VAS)and quantitative sensory testing(QST).The QST employed in this study included warm detection threshold/heat pain threshold(WDT/HPT),cold detection threshold/cold pain threshold(CDT/CPT),and pressure pain threshold(PPT).Tests were conducted on teeth 21 and 31,their gums,the left hand,and the left masseter muscle.Assessments occurred at baseline,2 hours,24 hours,7 days,and 28 days after aligner placement.Re-sults In the experimental group,VAS scores peaked at 24 hours and then significantly decreased(P＜0.01).For teeth 21 and 31,the WDT and HPT of the labial attached gingiva,along with the PPT of these teeth,showed significant decreases at 24 hours and 7 days compared to baseline and the control group(P＜0.01).At 24 hours and 7 days,VAS scores were negatively correlated with the WDT and HPT of the labial attached gingiva of teeth 21 and 31(P＜0.05).Conclusion At 24 hours and 7 days post-clear aligner treatment,thermal sensitivity and pain,as well as pressure pain sensitivity,increased for the gingiva of teeth 21 and 31.Thermal sensitivity was also correlated with pain intensity.

Cervical spinal cord injury without fracture-dislocation (CSCIWFD) is referred to as a special type of cervical spinal cord injury characterized by traumatic spinal cord dysfunction and no significant bony structural abnormalities on imagines. Duo to the high risk of missed diagnosis during the initial consultation, CSCIWFD may lead to progressive neurological deterioration or even complete paralysis, severely impacting patients′ prognosis. Currently, there are no established consensuses over the diagnosis and treatment of CSCIWFD, such as the lack of evidence-based standards for indications of non-surgical treatment and risk of secondary neurological injury, as well as debates over the optimal timing for surgical intervention and indications for different surgical approaches. To address these issues, the Spine Trauma Group of the Orthopedic Branch of the Chinese Medical Doctor Association organized experts in the relevant fields to formulate Diagnosis and treatment guideline for acute cervical spinal cord injury without fracture- dislocation in adults ( version 2025) . Based on evidence-based medicine and the principles of scientific rigor and clinical applicability, the guidelines proposed 11 recommendations covering terminology, diagnosis, evaluation treatment, and rehabilitation, etc., aiming to standardize the management of CSCIWFD.

Objective To explore the changes in neural activity in patients with type 2 diabetes mellitus(T2DM)and their corre-lation with executive function,and to analyze the neural mechanisms underlying the decline in executive function in T2DM patients.Methods Thirty-one T2DM patients(T2DM group)and thirty-two healthy controls(HC)(HC group)matched for body mass index(BMI)underwent resting-state functional magnetic resonance imaging(rs-fMRI)scans and N-back task tests were included.Differ-ences in the amplitude of low-frequency fluctuation(ALFF),regional homogeneity(ReHo),and seed-based functional connectivity(FC)between the two groups were compared,and partial correlation analyses were performed between the difference results and N-back task performance.Results The T2DM group showed prolonged reaction time(RT)in the 1-back and 2-back tasks.T2DM patients exhibited increased ALFF in the bilateral caudate nucleus,left medial superior frontal gyrus,and right postcentral gyrus,as well as elevated ReHo in the right putamen.FC analysis revealed significant alterations in FC between the caudate nucleus,putamen,and multiple brain regions in T2DM patients,with some of these FC changes significantly correlated with RT and accuracy(ACC)in the N-back task.Conclusion The decline in executive function in T2DM patients may be associated with abnormal neural activity in brain regions such as the striatum,salience network,and frontoparietal control network.FC further decreases under increased cognitive load.These findings provide evidence for the study of the neural mechanisms of executive function impairment in T2DM patients.

Objective To investigate the initial temperature sensation and pressure pain sensation in patients receiving invisible or-thodontic treatment.Methods Twenty-two patients receiving clear aligner treatment(experimental group)and 22 volunteers(control group)participated.Perceptual changes were assessed using the visual analogue scale(VAS)and quantitative sensory testing(QST).The QST employed in this study included warm detection threshold/heat pain threshold(WDT/HPT),cold detection threshold/cold pain threshold(CDT/CPT),and pressure pain threshold(PPT).Tests were conducted on teeth 21 and 31,their gums,the left hand,and the left masseter muscle.Assessments occurred at baseline,2 hours,24 hours,7 days,and 28 days after aligner placement.Re-sults In the experimental group,VAS scores peaked at 24 hours and then significantly decreased(P＜0.01).For teeth 21 and 31,the WDT and HPT of the labial attached gingiva,along with the PPT of these teeth,showed significant decreases at 24 hours and 7 days compared to baseline and the control group(P＜0.01).At 24 hours and 7 days,VAS scores were negatively correlated with the WDT and HPT of the labial attached gingiva of teeth 21 and 31(P＜0.05).Conclusion At 24 hours and 7 days post-clear aligner treatment,thermal sensitivity and pain,as well as pressure pain sensitivity,increased for the gingiva of teeth 21 and 31.Thermal sensitivity was also correlated with pain intensity.