OBJECTIVE To investigate the acute lung injury effects of pulmonary phospholipids and their phospholipase A2(PLA2)decomposition products-lysophospholipids and fatty acids-on mice.METHODS Mice were randomly assigned to the following groups:① solvent control(PBS)and PLA2;② solvent control and glycerol phospholipid groups:1,2-dioleoyl-sn-glycero-3-phosphoserine(DOPS),1,2-dipalmitoyl-sn-glycero-3-phosphoserine(DPPS),1,2-dioleoyl-sn-glycero-3-phosphoethanol-amine(DOPE),1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine(DPPE),1,2-dipalmitoyl-sn-glycero-3-phosphocholine(DPPC),and 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine(SOPC);③ solvent con-trol and fatty acid groups:palmitic acid(PA),oleic acid;④ solvent control and lysophospholipid groups:1-oleoyl-2-hydroxy-sn-glycero-3-phosphoserine(18∶1 LysoPS),1-stearoyl-sn-glycero-3-phosphoserine(18∶0 LysoPS),1-palmitoyl-sn-glycero-3-phosphoserine(16∶0 LysoPS),1-palmitoyl-sn-glycero-3-phos-phoethanolamine(16∶0 LysoPE),1-palmitoyl-sn-glycero-3-phosphocholine(16∶0 LysoPC);⑤ solvent control,PLA2,DPPC,PA,16∶0 LysoPC,16∶0 LysoPS,and 18∶1 LysoPS.Following anesthesia,mice were administered nebulized PBS in the solvent control group,2.1 ug·kg-1 PLA2 in PBS in the PLA2 group and 2.5 mg·kg-1 of the corresponding substance in PBS in other experimental groups.For group①,survival times were recorded and survival curves were plotted.At 1 h post-treatment,lung tissues from groups ①②③④ were collected,photographed to obtain white light images,and subjected to HE staining to assess histopathological changes and pathological scoring.At 2 h post-treatment,pulmonary blood flow in group ⑤ was assessed using laser speckle contrast imaging,arterial blood gas was analyzed with a blood gas analyzer,and lung function was evaluated using whole-body pleth-ysmography.At 6 hours post-treatment,blood cells from group ⑤ were analyzed using an automated hematology analyzer.RESULTS Compared with the solvent control group,severe pathological changes were observed in lung tissues of the PLA2 group,accompanied by extensive inflammatory infiltration and interstitial thickening,with all mice succumbing within 240 min.In mice treated with glyc-erol phospholipids,alveolar structures remained clear,alveolar walls were intact and continuous,and alveolar spaces were translucent,with only occasional minor inflammatory cell infiltration in the septa.No significant pathological alterations were detected in the fatty acid groups.Minor inflammatory cell infiltration was seen in the 16∶0 LysoPE and 16∶0 LysoPC groups.However,such pathological changes as patchy hemorrhage,alveolar interstitial edema,increased alveolar wall thickness,and elevated neutrophil counts were observed in the 18∶1 LysoPS,18∶0 LysoPS,and 16∶0 LysoPS groups.Pathological scores based on HE staining were significantly increased in the 16∶0 LysoPS and 18∶1 LysoPS groups com-pared with the solvent control.The percentage of the lung tissue injury area was also markedly higher in the 16∶0 LysoPS group.A significant decrease in the mean fluorescence intensity of blood flow was observed in the 16∶0 LysoPS group.Arterial partial pressure of oxygen(pO2)was significantly reduced in the PLA2 group,while arterial partial pressure of carbon dioxide(pCO2)was markedly elevated in the 16∶0 LysoPS and 18∶1 LysoPS groups.Lung function tests revealed that the 16∶0 LysoPS group exhibited significant increases in expiratory time,end-expiratory pressure,and enhanced pause,in contrast to significant decreases in tidal volume,expired volume,and minute volume.The 18∶1 LysoPS group also exhibited a significant decline in minute volume.No significant changes in inflammatory cell concentrations were detected in blood,with the exception of neutrophils in the 16∶0 LysoPS group,which showed a significant but physiologically normal increase.CONCLUSION Pulmonary phospholipids and their PLA2-derived fatty acid metabolites do not induce severe lung injury in mice while the lyso-phospholipid metabolites,particularly lysophosphatidylserine,are found to cause significant lung injury.

Objective To compare the macular retinal thickness characteristics of diabetic kidney disease(DKD)patients with different blood pressure levels and explore the relationship between blood pressure and macular retinal damage in DKD patients.Methods A total of 435 DKD patients were selected and divided into four groups based on medical history and blood pressure:the non-hypertensive group(NH group,n=100),the well-controlled blood pressure group(G0 group,n=176),the grade 1 hypertension group(G1 group,n=118)and the grade 2 hypertension group(G2 group,n=41).General information,routine laboratory test results as well as the average thickness of the macular retinal nerve fiber layer(RNFL),ganglion cell layer(GCL),ganglion cell complex(GCC),choroidal layer(CL)and central macular retinal thickness(CMT)were compared between the four groups.The correlation between macular retinal thickness in both eyes and clinical data was analyzed.Results Compared with the G1 group and the G2 group,the G0 group had a longer duration of hypertension.Compared to the NH group,the G2 group had higher fasting plasma glucose(FPG).Compared to the G0 group,the G1 group and the G2 group had higher FPG,and the G2 group had higher glycated hemoglobin(HbA1c)and lower estimated glomerular filtration rate(eGFR).Compared to the NH group,the G0 group had decreased thickness in the GCL and GCC(P<0.05).The macular retinal thickness of the GCL and GCC in both eyes was negatively correlated with diastolic blood pressure(r=-0.158 and-0.195,respectively,P<0.05).Conclusion Macular retinal thickness is helpful in assessing the long-term effects of hypertension on optic nerve and microvascular damage in DKD patients.

Objective To investigate serum metabolites and metabolic pathways alterations in patients with ischemic stroke(IS)through metabolomic analysis,and to identify reliable serum metabolic biomarkers for IS diagnosis.Methods This prospective study enrolled patients with IS admitted to the Department of Neurology at Xiangcheng People's Hospital of Suzhou from December 1,2022 to December 31,2023.Age-and sex-matched healthy individuals were recruited as controls during the same period.Baseline characteristics were collected,including age,sex,height,body mass index,and blood pressure.Venous blood samples were obtained after an 8 h fast for biochemical analysis of blood glucose,total bilirubin,serum creatinine,urea nitrogen,total cholesterol,triglycerides,high-density lipoprotein cholesterol,and low-density lipoprotein cholesterol.Serum metabolites of both groups were extracted and analyzed using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry.Metabolomic data were processed using Simca-p software for unsupervised principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)to evaluate group separation and experimental stability.Differential metabolites were defined by variable importance in projection(VIP)≥1.0,fold change(FC)≥2.0 or ≤0.5,and P＜0.05.Drug-derived exogenous metabolites were excluded by cross-referencing the Human Metabolome Database(HMDB,https://hmdb.ca/)and PubChem(https://pubchem.ncbi.nlm.nih.gov/).MetaboAnalyst 6.0(http://www.metaboanalyst.ca),a comprehensive web-based tool for metabolomic data analysis,was employed to map differential metabolites to the Kyoto encyclopedia of genes and genomes(KEGG)databased and to perform pathway enrichment analysis.Machine learning models were developed using Python.Least absolute shrinkage and selection operator(LASSO)regression and random forest(RF)algorithms were employed to identify diagnostic biomarkers capable of effectively distinguishing IS patients from controls.Metabolites identified by both methods were integrated into an extreme gradient boosting(XGBoost)model.Model performance was evaluated using receiver operating characteristic(ROC)curves with 5-fold cross-validation and internal validation(70％training,30％validation set).Results A total of 51 IS patients and 51 matched controls were included.(1)A total of 1 255 serum metabolites were identified(964 in positive ion mode,291 in negative ion mode).PC A and OPLS-DA demonstrated distinct metabolic separation between IS patients and controls.In IS group,260 metabolites were upregulated and 337 downregulated in positive ion mode;99 were upregulated and 34downregulated in negative ion mode.(2)Among the 1 255 metabolites,259 were identified as differential metabolites based on the criteria of VIP ≥ 1.0,FC≥2.0 or≤0.5 and P＜0.05.After excluding drug-derived metabolite through referencing HMDB and PubChem databases,a total of 220 endogenous differential metabolites were found to coexist in both positive and negative ion modes.Among them,119 metabolites were up-regulated and 101 were down-regulated in the IS group.The expression of these 220 metabolites showed significant differences between the IS and control groups.(3)KEGG pathway analysis highlighted five dysregulated pathways:upregulation of denovo triacylglycerol biosynthesis,glycerophosphate shuttle,and cardiolipin biosynthesis;downregulation of bile acid biosynthesis and methylhistidine metabolism.(4)LASSO and RF algorithms identified 24 and 30 candidate biomarkers,respectively.Four overlapping metabolites were selected:2-((3R)-3-((3R,5S,7S,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)butanamido)ethane-1-sulfonic acid(m/z 971.571 29),arginine-conjugated cholic acid(m/z 587.379 21),laccaic acid A(m/z 576.010 93)and NCGC00380235-01_C32H48O9_beta-D-xylopyranoside,3,17-dihydroxyspirosta-5,25(27)-dien-1-yl(m/z 559.326 48).The expression levels of 2-((3R)-3-((3R,5S,7S,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)butanamido)ethane-1-sulfonic acid(m/z 971.571 29),arginine-conjugated cholic acid(m/z 587.379 21),and laccaic acid A(m/z 576.010 93)were upregulated,while the expression level of NCGC00380235-01_C32H48O9_beta-D-xylopyranoside,3,17-dihydroxyspirosta-5,25(27)-dien-1-yl(m/z 559.326 48)was downregulated.An IS diagnostic model was established based on four metabolic biomarkers using the XGBoost algorithm.The area under the ROC curve was 1.000(95％CI 1.000-1.000)in the training set and 0.988 in the validation set(95％CI 0.963-1.000).Conclusions Patients with IS exhibit significant metabolic disturbance.The four identified biomarkers may serve as potential biomarkers for the effective identification of IS:2-((3 R)-3-((3R,5S,7S,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)butanamido)ethane-1-sulfonic acid(m/z971.571 29),arginine-conjugated cholic acid(m/z587.379 21),laccaic acid A(m/z 576.010 93),and NCGC00380235-01_C32H48O9_beta-D-xylopyranoside,3,17-dihydroxyspirosta-5,25(27)-dien-1-yl(m/z 559.326 48).

OBJECTIVE To construct novel central nervous system(CNS)-targeted lipid nanoparti-cles for the treatment of organophosphorus-induced brain injury in mice.METHODS(1)Preparation,screening,and characterization of lipid nanoparticles.① Lipid nanoreactivators were prepared using the thin-film hydration method,with asoxime(HI-6)as the therapeutic drug and lipid carriers composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine(POPS),1,2-dipalmitoyl-sn-glycero-3-phospho-choline(DPPC),and cholesterol(CHOL)(PDC)at varying molar ratios(1∶6∶3,3∶4∶3,5∶2∶3 and 7∶0∶3)(HI-6@PDC 1∶6∶3,3∶4∶3,5∶2∶3 and 7∶0∶3).② FLU-labeled lipid nanocarriers(FLU@PDC 1∶6∶3,3∶4∶3,5∶2∶3,and 7∶0∶3)were prepared and physically mixed with phospholipase A2(PLA2)solution(at the final PLA2 concentration of 10 kU·L-1)to obtain FLU@PDC+PLA2.Male KM mice were randomly divided into normal control(PBS),FLU,and FLU@PDC+PLA2(1∶6∶3,3∶4∶3,5∶2∶3,and 7∶0∶3)groups(n=7 per group).After intravenous(iv)administration(FLU dose:1 mg·kg-1,carrier dose:80 mg·kg-1),brain tissues were collected at 1 h,homogenized,centrifuged,and analyzed via fluorescence spectrophotom-etry to screen the optimal CNS-targeted lipid carrier composition.③ The morphology of HI-6@PDC 5∶2∶3 was characterized by transmission electron microscope(TEM).The particle size,polydispersity index(PDI),and zeta potential of HI-6@PDC 5∶2∶3 were measured using a Zeta potential and particle size analyzer.Encapsulation efficiency and loading efficiency of HI-6@PDC 5∶2∶3 were determined using an ultrafiltration centrifugation method combined with high-performance liquid chromatography(HPLC).In vitro release kinetics of HI-6@PDC 5∶2∶3 and HI-6@PDC+PLA2 5∶2∶3 were assessed using a dialysis bag diffusion method combined with fluorescence spectrophotometry.(2)Validation of CNS targeting.① Cyanine7(Cy7)-labeled PDC 5∶2∶3(Cy7@PDC)was prepared and mixed with PLA2 solution(Cy7@PDC+PLA2 5∶2∶3).Mice were divided into normal control,Cy7,Cy7@PDC 5∶2∶3 and Cy7@PDC+PLA2 5∶2∶3 groups(n=3 per group).After iv injection(Cy7 dose:1 mg·kg-1,carrier dose:80 mg·kg-1),brain fluorescence was visualized at 3 h using a small animal in vivo imaging(IVIS)system.② Cyanine 3(Cy3)-labeled PDC 5∶2∶3(Cy3@PDC 5∶2∶3)was prepared and mixed with PLA2 solution(Cy3@PDC+PLA2 5∶2∶3).Mice were divided into Cy3@PDC 5∶2∶3 and Cy3@PDC+PLA2 5∶2∶3 groups(n=3 per group).After iv injection(Cy3 dose:1 mg·kg-1,carrier dose:80 mg·kg-1),brain tissues were collected at 2 h for fluorescent staining and Cy3 fluorescence observation.(3)Therapeutic efficacy eval-uation.① Male KM mice were randomly divided into normal control,brain injury,HI-6 treatment,and HI-6@PDC+PLA2 5∶2∶3 treatment groups(n=6 per group).Except for the normal control,all the mice were subcutaneously(sc)injected with soman(120 μg·kg-1),followed by immediate iv treatment(HI-6 dose:22 mg·kg-1,carrier dose:80 mg·kg-1).At 10 min,orbital blood and brain tissues were collected before brain weight was recorded.Acetylcholinesterase(AChE)reactivation in blood and brain was measured using the Ellman method.② Grouping and treatment were identical to ①(n=3 per group).At 24 h,brain tissues were collected for HE staining to assess histopathological damage.③ Mice were divided into brain injury and HI-6@PDC+PLA2 5∶2∶3 treatment groups(n=10 per group)and treated as in ①(soman dose:220 ug·kg-1).Survival rates,neurotoxic symptoms(tremors,salivation),and seizure latency were recorded,and survival curves were plotted.RESULTS(1)PDC 5∶2∶3 exhibited the highest brain fluorescence,indicating optimal CNS targeting.HI-6@PDC 5∶2∶3 appeared in regular spherical shapes,and were negatively charged,with a size of(219.4±3.1)nm,PDI of 0.4±0.02,entrapment effi-ciency of 72.9％and loading efficiency of 49.7％.HI-6@PDC+PLA2 5∶2∶3 showed a cumulative release of 43.5％at 60 min,which was lower than that of rhodamine B(RB)but sufficient for CNS therapeutic timelines.(2)In vivo fluorescence and pathological fluorescence confirmed PLA2-mediated CNS delivery.(3)HI-6@PDC+PLA2 5∶2∶3 significantly enhanced AChE reactivation in the blood and brain compared to HI-6.Histopathology revealed mitigated brain injury in treated mice.HI-6@PDC+PLA2 5∶2∶3 prolonged survival,reduced convulsions,alleviated neurotoxicity,and extended seizure latency.CONCLUSION HI-6@PDC 5∶2∶3 can effectively cross the blood-brain barrier via PLA2 mediation,demonstrating strong CNS targeting.It can significantly improve AChE reactivation in peripheral and central tissues and offers potent therapeutic efficacy against organophosphate-induced brain injury.

OBJECTIVE To construct novel central nervous system(CNS)-targeted lipid nanoparti-cles for the treatment of organophosphorus-induced brain injury in mice.METHODS(1)Preparation,screening,and characterization of lipid nanoparticles.① Lipid nanoreactivators were prepared using the thin-film hydration method,with asoxime(HI-6)as the therapeutic drug and lipid carriers composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine(POPS),1,2-dipalmitoyl-sn-glycero-3-phospho-choline(DPPC),and cholesterol(CHOL)(PDC)at varying molar ratios(1∶6∶3,3∶4∶3,5∶2∶3 and 7∶0∶3)(HI-6@PDC 1∶6∶3,3∶4∶3,5∶2∶3 and 7∶0∶3).② FLU-labeled lipid nanocarriers(FLU@PDC 1∶6∶3,3∶4∶3,5∶2∶3,and 7∶0∶3)were prepared and physically mixed with phospholipase A2(PLA2)solution(at the final PLA2 concentration of 10 kU·L-1)to obtain FLU@PDC+PLA2.Male KM mice were randomly divided into normal control(PBS),FLU,and FLU@PDC+PLA2(1∶6∶3,3∶4∶3,5∶2∶3,and 7∶0∶3)groups(n=7 per group).After intravenous(iv)administration(FLU dose:1 mg·kg-1,carrier dose:80 mg·kg-1),brain tissues were collected at 1 h,homogenized,centrifuged,and analyzed via fluorescence spectrophotom-etry to screen the optimal CNS-targeted lipid carrier composition.③ The morphology of HI-6@PDC 5∶2∶3 was characterized by transmission electron microscope(TEM).The particle size,polydispersity index(PDI),and zeta potential of HI-6@PDC 5∶2∶3 were measured using a Zeta potential and particle size analyzer.Encapsulation efficiency and loading efficiency of HI-6@PDC 5∶2∶3 were determined using an ultrafiltration centrifugation method combined with high-performance liquid chromatography(HPLC).In vitro release kinetics of HI-6@PDC 5∶2∶3 and HI-6@PDC+PLA2 5∶2∶3 were assessed using a dialysis bag diffusion method combined with fluorescence spectrophotometry.(2)Validation of CNS targeting.① Cyanine7(Cy7)-labeled PDC 5∶2∶3(Cy7@PDC)was prepared and mixed with PLA2 solution(Cy7@PDC+PLA2 5∶2∶3).Mice were divided into normal control,Cy7,Cy7@PDC 5∶2∶3 and Cy7@PDC+PLA2 5∶2∶3 groups(n=3 per group).After iv injection(Cy7 dose:1 mg·kg-1,carrier dose:80 mg·kg-1),brain fluorescence was visualized at 3 h using a small animal in vivo imaging(IVIS)system.② Cyanine 3(Cy3)-labeled PDC 5∶2∶3(Cy3@PDC 5∶2∶3)was prepared and mixed with PLA2 solution(Cy3@PDC+PLA2 5∶2∶3).Mice were divided into Cy3@PDC 5∶2∶3 and Cy3@PDC+PLA2 5∶2∶3 groups(n=3 per group).After iv injection(Cy3 dose:1 mg·kg-1,carrier dose:80 mg·kg-1),brain tissues were collected at 2 h for fluorescent staining and Cy3 fluorescence observation.(3)Therapeutic efficacy eval-uation.① Male KM mice were randomly divided into normal control,brain injury,HI-6 treatment,and HI-6@PDC+PLA2 5∶2∶3 treatment groups(n=6 per group).Except for the normal control,all the mice were subcutaneously(sc)injected with soman(120 μg·kg-1),followed by immediate iv treatment(HI-6 dose:22 mg·kg-1,carrier dose:80 mg·kg-1).At 10 min,orbital blood and brain tissues were collected before brain weight was recorded.Acetylcholinesterase(AChE)reactivation in blood and brain was measured using the Ellman method.② Grouping and treatment were identical to ①(n=3 per group).At 24 h,brain tissues were collected for HE staining to assess histopathological damage.③ Mice were divided into brain injury and HI-6@PDC+PLA2 5∶2∶3 treatment groups(n=10 per group)and treated as in ①(soman dose:220 ug·kg-1).Survival rates,neurotoxic symptoms(tremors,salivation),and seizure latency were recorded,and survival curves were plotted.RESULTS(1)PDC 5∶2∶3 exhibited the highest brain fluorescence,indicating optimal CNS targeting.HI-6@PDC 5∶2∶3 appeared in regular spherical shapes,and were negatively charged,with a size of(219.4±3.1)nm,PDI of 0.4±0.02,entrapment effi-ciency of 72.9％and loading efficiency of 49.7％.HI-6@PDC+PLA2 5∶2∶3 showed a cumulative release of 43.5％at 60 min,which was lower than that of rhodamine B(RB)but sufficient for CNS therapeutic timelines.(2)In vivo fluorescence and pathological fluorescence confirmed PLA2-mediated CNS delivery.(3)HI-6@PDC+PLA2 5∶2∶3 significantly enhanced AChE reactivation in the blood and brain compared to HI-6.Histopathology revealed mitigated brain injury in treated mice.HI-6@PDC+PLA2 5∶2∶3 prolonged survival,reduced convulsions,alleviated neurotoxicity,and extended seizure latency.CONCLUSION HI-6@PDC 5∶2∶3 can effectively cross the blood-brain barrier via PLA2 mediation,demonstrating strong CNS targeting.It can significantly improve AChE reactivation in peripheral and central tissues and offers potent therapeutic efficacy against organophosphate-induced brain injury.

OBJECTIVE To investigate the acute lung injury effects of pulmonary phospholipids and their phospholipase A2(PLA2)decomposition products-lysophospholipids and fatty acids-on mice.METHODS Mice were randomly assigned to the following groups:① solvent control(PBS)and PLA2;② solvent control and glycerol phospholipid groups:1,2-dioleoyl-sn-glycero-3-phosphoserine(DOPS),1,2-dipalmitoyl-sn-glycero-3-phosphoserine(DPPS),1,2-dioleoyl-sn-glycero-3-phosphoethanol-amine(DOPE),1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine(DPPE),1,2-dipalmitoyl-sn-glycero-3-phosphocholine(DPPC),and 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine(SOPC);③ solvent con-trol and fatty acid groups:palmitic acid(PA),oleic acid;④ solvent control and lysophospholipid groups:1-oleoyl-2-hydroxy-sn-glycero-3-phosphoserine(18∶1 LysoPS),1-stearoyl-sn-glycero-3-phosphoserine(18∶0 LysoPS),1-palmitoyl-sn-glycero-3-phosphoserine(16∶0 LysoPS),1-palmitoyl-sn-glycero-3-phos-phoethanolamine(16∶0 LysoPE),1-palmitoyl-sn-glycero-3-phosphocholine(16∶0 LysoPC);⑤ solvent control,PLA2,DPPC,PA,16∶0 LysoPC,16∶0 LysoPS,and 18∶1 LysoPS.Following anesthesia,mice were administered nebulized PBS in the solvent control group,2.1 ug·kg-1 PLA2 in PBS in the PLA2 group and 2.5 mg·kg-1 of the corresponding substance in PBS in other experimental groups.For group①,survival times were recorded and survival curves were plotted.At 1 h post-treatment,lung tissues from groups ①②③④ were collected,photographed to obtain white light images,and subjected to HE staining to assess histopathological changes and pathological scoring.At 2 h post-treatment,pulmonary blood flow in group ⑤ was assessed using laser speckle contrast imaging,arterial blood gas was analyzed with a blood gas analyzer,and lung function was evaluated using whole-body pleth-ysmography.At 6 hours post-treatment,blood cells from group ⑤ were analyzed using an automated hematology analyzer.RESULTS Compared with the solvent control group,severe pathological changes were observed in lung tissues of the PLA2 group,accompanied by extensive inflammatory infiltration and interstitial thickening,with all mice succumbing within 240 min.In mice treated with glyc-erol phospholipids,alveolar structures remained clear,alveolar walls were intact and continuous,and alveolar spaces were translucent,with only occasional minor inflammatory cell infiltration in the septa.No significant pathological alterations were detected in the fatty acid groups.Minor inflammatory cell infiltration was seen in the 16∶0 LysoPE and 16∶0 LysoPC groups.However,such pathological changes as patchy hemorrhage,alveolar interstitial edema,increased alveolar wall thickness,and elevated neutrophil counts were observed in the 18∶1 LysoPS,18∶0 LysoPS,and 16∶0 LysoPS groups.Pathological scores based on HE staining were significantly increased in the 16∶0 LysoPS and 18∶1 LysoPS groups com-pared with the solvent control.The percentage of the lung tissue injury area was also markedly higher in the 16∶0 LysoPS group.A significant decrease in the mean fluorescence intensity of blood flow was observed in the 16∶0 LysoPS group.Arterial partial pressure of oxygen(pO2)was significantly reduced in the PLA2 group,while arterial partial pressure of carbon dioxide(pCO2)was markedly elevated in the 16∶0 LysoPS and 18∶1 LysoPS groups.Lung function tests revealed that the 16∶0 LysoPS group exhibited significant increases in expiratory time,end-expiratory pressure,and enhanced pause,in contrast to significant decreases in tidal volume,expired volume,and minute volume.The 18∶1 LysoPS group also exhibited a significant decline in minute volume.No significant changes in inflammatory cell concentrations were detected in blood,with the exception of neutrophils in the 16∶0 LysoPS group,which showed a significant but physiologically normal increase.CONCLUSION Pulmonary phospholipids and their PLA2-derived fatty acid metabolites do not induce severe lung injury in mice while the lyso-phospholipid metabolites,particularly lysophosphatidylserine,are found to cause significant lung injury.

Objective To compare the macular retinal thickness characteristics of diabetic kidney disease(DKD)patients with different blood pressure levels and explore the relationship between blood pressure and macular retinal damage in DKD patients.Methods A total of 435 DKD patients were selected and divided into four groups based on medical history and blood pressure:the non-hypertensive group(NH group,n=100),the well-controlled blood pressure group(G0 group,n=176),the grade 1 hypertension group(G1 group,n=118)and the grade 2 hypertension group(G2 group,n=41).General information,routine laboratory test results as well as the average thickness of the macular retinal nerve fiber layer(RNFL),ganglion cell layer(GCL),ganglion cell complex(GCC),choroidal layer(CL)and central macular retinal thickness(CMT)were compared between the four groups.The correlation between macular retinal thickness in both eyes and clinical data was analyzed.Results Compared with the G1 group and the G2 group,the G0 group had a longer duration of hypertension.Compared to the NH group,the G2 group had higher fasting plasma glucose(FPG).Compared to the G0 group,the G1 group and the G2 group had higher FPG,and the G2 group had higher glycated hemoglobin(HbA1c)and lower estimated glomerular filtration rate(eGFR).Compared to the NH group,the G0 group had decreased thickness in the GCL and GCC(P<0.05).The macular retinal thickness of the GCL and GCC in both eyes was negatively correlated with diastolic blood pressure(r=-0.158 and-0.195,respectively,P<0.05).Conclusion Macular retinal thickness is helpful in assessing the long-term effects of hypertension on optic nerve and microvascular damage in DKD patients.

Objective To investigate serum metabolites and metabolic pathways alterations in patients with ischemic stroke(IS)through metabolomic analysis,and to identify reliable serum metabolic biomarkers for IS diagnosis.Methods This prospective study enrolled patients with IS admitted to the Department of Neurology at Xiangcheng People's Hospital of Suzhou from December 1,2022 to December 31,2023.Age-and sex-matched healthy individuals were recruited as controls during the same period.Baseline characteristics were collected,including age,sex,height,body mass index,and blood pressure.Venous blood samples were obtained after an 8 h fast for biochemical analysis of blood glucose,total bilirubin,serum creatinine,urea nitrogen,total cholesterol,triglycerides,high-density lipoprotein cholesterol,and low-density lipoprotein cholesterol.Serum metabolites of both groups were extracted and analyzed using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry.Metabolomic data were processed using Simca-p software for unsupervised principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)to evaluate group separation and experimental stability.Differential metabolites were defined by variable importance in projection(VIP)≥1.0,fold change(FC)≥2.0 or ≤0.5,and P＜0.05.Drug-derived exogenous metabolites were excluded by cross-referencing the Human Metabolome Database(HMDB,https://hmdb.ca/)and PubChem(https://pubchem.ncbi.nlm.nih.gov/).MetaboAnalyst 6.0(http://www.metaboanalyst.ca),a comprehensive web-based tool for metabolomic data analysis,was employed to map differential metabolites to the Kyoto encyclopedia of genes and genomes(KEGG)databased and to perform pathway enrichment analysis.Machine learning models were developed using Python.Least absolute shrinkage and selection operator(LASSO)regression and random forest(RF)algorithms were employed to identify diagnostic biomarkers capable of effectively distinguishing IS patients from controls.Metabolites identified by both methods were integrated into an extreme gradient boosting(XGBoost)model.Model performance was evaluated using receiver operating characteristic(ROC)curves with 5-fold cross-validation and internal validation(70％training,30％validation set).Results A total of 51 IS patients and 51 matched controls were included.(1)A total of 1 255 serum metabolites were identified(964 in positive ion mode,291 in negative ion mode).PC A and OPLS-DA demonstrated distinct metabolic separation between IS patients and controls.In IS group,260 metabolites were upregulated and 337 downregulated in positive ion mode;99 were upregulated and 34downregulated in negative ion mode.(2)Among the 1 255 metabolites,259 were identified as differential metabolites based on the criteria of VIP ≥ 1.0,FC≥2.0 or≤0.5 and P＜0.05.After excluding drug-derived metabolite through referencing HMDB and PubChem databases,a total of 220 endogenous differential metabolites were found to coexist in both positive and negative ion modes.Among them,119 metabolites were up-regulated and 101 were down-regulated in the IS group.The expression of these 220 metabolites showed significant differences between the IS and control groups.(3)KEGG pathway analysis highlighted five dysregulated pathways:upregulation of denovo triacylglycerol biosynthesis,glycerophosphate shuttle,and cardiolipin biosynthesis;downregulation of bile acid biosynthesis and methylhistidine metabolism.(4)LASSO and RF algorithms identified 24 and 30 candidate biomarkers,respectively.Four overlapping metabolites were selected:2-((3R)-3-((3R,5S,7S,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)butanamido)ethane-1-sulfonic acid(m/z 971.571 29),arginine-conjugated cholic acid(m/z 587.379 21),laccaic acid A(m/z 576.010 93)and NCGC00380235-01_C32H48O9_beta-D-xylopyranoside,3,17-dihydroxyspirosta-5,25(27)-dien-1-yl(m/z 559.326 48).The expression levels of 2-((3R)-3-((3R,5S,7S,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)butanamido)ethane-1-sulfonic acid(m/z 971.571 29),arginine-conjugated cholic acid(m/z 587.379 21),and laccaic acid A(m/z 576.010 93)were upregulated,while the expression level of NCGC00380235-01_C32H48O9_beta-D-xylopyranoside,3,17-dihydroxyspirosta-5,25(27)-dien-1-yl(m/z 559.326 48)was downregulated.An IS diagnostic model was established based on four metabolic biomarkers using the XGBoost algorithm.The area under the ROC curve was 1.000(95％CI 1.000-1.000)in the training set and 0.988 in the validation set(95％CI 0.963-1.000).Conclusions Patients with IS exhibit significant metabolic disturbance.The four identified biomarkers may serve as potential biomarkers for the effective identification of IS:2-((3 R)-3-((3R,5S,7S,9S,10S,13R,14S,17R)-3,7-dihydroxy-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-17-yl)butanamido)ethane-1-sulfonic acid(m/z971.571 29),arginine-conjugated cholic acid(m/z587.379 21),laccaic acid A(m/z 576.010 93),and NCGC00380235-01_C32H48O9_beta-D-xylopyranoside,3,17-dihydroxyspirosta-5,25(27)-dien-1-yl(m/z 559.326 48).

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To explore the clinical phenotype and pathogenesis of a child with partial duplication in the long arm of chromosome 10 (10q), and conduct a review of relevant literature.Methods:A child presented at Lianyungang Maternal and Child Health Care Hospital in April 2018 for growth retardation, intellectual disability, and autism spectrum disorder (ASD) was selected as the study subject. Peripheral blood samples were collected from the child and his parents for G-banded chromosomal karyotyping analysis. Genomic DNA was also extracted for chromosomal microarray analysis (CMA). The clinical phenotype and relevant genes were searched in the Online Mendelian Inheritance in Man (OMIM) and the UK Database of Genomic Variation and Phenotype in Humans using Ensembl Resources (DECIPHER). The pathogenicity of chromosomal variation was analyzed based on guidelines from the American College of Medical Genetics and Genomics (ACMG). Relevant literature was searched from the CNKI, Wanfang Data, and PubMed databases by using keywords such as " 10q" " duplication" and " trisomy", with the time set as from the establishment of database to December 1, 2023. This study has been approved by the Medical Ethics Committee of the Lianyungang Maternal and Child Health Care Hospital (No. XM2023030).Results:The clinical phenotype of child had included growth retardation, intellectual disability, and ASD. G-banded chromosomal analysis suggested that the child has a karyotype of 46, XY, dup(10)(q23.31q24.33), whilst both of his parents were normal. CMA analysis of the child revealed that the child was arr[19]10q23.31q24.33(87603382_104948862)×3, with a 17.34 Mb duplication in the 10q23.31q24.33 region. Search of the OMIM database suggested that the duplicated segment has contained 171 genes associated with various diseases, and search of the DECIPHER database has identified cases with overlapping with the duplication. A search of the PubMed database has identified 2 publications involving 2 patients with chromosomal duplications overlapping the 10q23.31q24.33 region with a segment length of > 10 Mb. The 2 patients had mainly manifested growth retardation, intellectual disability, ASD, and facial and limb malformations. The main pathogenic genes had included PTEN, WNT8B, LZTS2, NFKB2, PAX2, KIF11, FRA10AC1, and CNNM2. No similar case was retrieved from the CNKI and Wanfang Data databases. Conclusion:The partial 10q duplication as a novel CNV involving genes such as PTEN and WNT8B probably underlay the growth retardation, intellectual disability and ASD in the child. This study has enriched the genotype-phenotype spectrum of patients with partial 10q23.31q24.33 duplications.