Objective To explore the protective effect of L-carnitine on myocardial systolic dysfunction in sepsis and its underlying mechanism.Methods A mouse sepsis model was established by cecal ligation and puncture(CLP).Ten-week-old male SPF-grade C57BL/6 mice(body weight 20～30 g)were randomly divided into 5 groups via random number table:Sham group,Sepsis group,L-carnitine group,L-carnitine+Etomoxir(Eto)group,and Eto group.Echocardiography assessed cardiac function,ELISA measured serum creatine kinase isoenzyme MB(CK-MB)levels,and 72-hour survival rates were recorded to evaluate L-carnitine's effects on cardiac function.Cardiomyocytes were isolated,and a cell microtensiometer was used to detect cardiomyocyte contractile function and calcium transients.Myocardial tissues were collected from each group,and ELISA was used to determine the contents of triglyceride(TG),free fatty acid(FFA),and adenosine triphosphate(ATP).An in vitro sepsis model was constructed by stimulating HL-1 cardiomyocytes with lipopolysaccharide(LPS)for 12 hours,which was divided into 5 groups:control(CTRL)group,LPS group,L-carnitine group,L-carnitine+Eto group,and Eto group.ELISA was used to detect the contents of TG,FFA,and ATP as well as the activity of carnitine palmitoyltransferase 1A(CPT1A)in cardiomyocytes.A cellular energy metabolism analysis system was employed to measure fatty acid oxidation capacity,and Western blot was used to detect the protein expression of CPT1A in cardiomyocytes.BODIPY-FL-C16(green fluorescently labeled palmitic acid)was utilized to detect the distribution of fatty acids in the cytoplasm and mitochondria via immunofluorescence technology,thereby observing the ability of cells to transport fatty acids into mitochondria.Results Compared with the Sham group,cardiac function was significantly impaired in the Sepsis group,as evidenced by decreased ejection fraction and mean arterial pressure(P<0.05),along with elevated levels of the cardiac injury marker CK-MB(P<0.05).Treatment with L-carnitine significantly improved myocardial function,restored blood pressure in septic mice,and increased their survival rate from 12.50%to 81.25%(P<0.05).Compared with the Sham group,the contractile function and calcium transients of acutely isolated single cardiomyocytes were significantly reduced in the Sepsis group(P<0.05),while L-carnitine treatment remarkably restored the contractile function and calcium release capacity of septic cardiomyocytes(P<0.05).Both in vivo and in vitro experiments showed that TG and FFA levels were significantly increased(P<0.05),and ATP levels was significantly decreased(P<0.05)in the Sepsis and LPS groups—effects significantly reversed by L-carnitine treatment.Compared with the CTRL group,the basal oxidation rate and maximum oxidation capacity of fatty acids in cardiomyocytes of the LPS group were significantly reduced(P<0.05),and L-carnitine treatment notably improved these indicators.Compared with the CTRL group,the expression and activity of CPT1A in cardiomyocytes of the LPS group were significantly decreased(P<0.05),while L-carnitine treatment significantly increased the expression and activity of CPT1A(P<0.05).In LPS group cardiomyocytes,green fluorescently labeled palmitic acid primarily formed numerous granular/clumpy aggregates in the cytoplasm with minimal mitochondrial colocalization.In the L-carnitine group,the green fluorescent granules in the cytoplasm of cardiomyocytes were smaller,and colocalization with mitochondria was increased.However,the L-carnitine+Eto group exhibited similar phenomena to the LPS group.In addition,both in vivo and in vitro experiments demonstrated that treatment with the CPT1A inhibitor Eto reversed the effect of L-carnitine.Compared with the L-carnitine group,the ATP content in the L-carnitine+Eto group was significantly decreased(P<0.05),while the FFA content was significantly increased(P<0.05).Conclusion L-carnitine facilitates fatty acid entry into mitochondria for β-oxidation via a CPT1A-dependent mechanism,thereby ameliorating fatty acid oxidation dysfunction in septic cardiomyocytes and improving myocardial contractile function.

Objective To explore the diagnostic value of serum C-C motif chemokine ligand 3(CCL3)and Semaphorin 3A in patients with type 2 diabetes mellitus(T2DM)complicated by osteoporosis(OP).Methods A total of 125 patients with T2DM complicated by OP who visited the hospital from August 2022 to August 2023 were selected as the OP group,and another 125 patients with T2DM without OP who visited the hospital during the same period were selected as the T2DM group.The OP group was divided into Group A(T value>-1.0,but slightly lower than the normal value),Group B(T value between-2.5 and-1.0),and Group C(T value<-2.5)based on the measurement results of bone mineral density(BMD).The lev-els of serum CCL3 and Semaphorin 3A in each group were compared.Pearson correlation analysis was used to analyze the correlations between serum CCL3,Semaphorin 3A and BMD in patients with T2DM complicated by OP.Multivariate Logistic regression analysis was used to analyze the influencing factors of T2DM compli-cated by OP.The receiver operating characteristic(ROC)curve was used to analyze the diagnostic value of se-rum CCL3 and Semaphorin 3A for T2DM complicated by OP.Results Compared with the T2DM group,the serum CCL3 level in the OP group increased,and the Semaphorin 3A level decreased,and the difference was statistically significant(P<0.05).Compared with group A,the serum CCL3 levels in group B and group C increased,while the Semaphorin 3A level decreased,and the differences were statistically significant(P<0.05).Compared with group B,the serum CCL3 level in group C increased and the Semaphorin 3A level de-creased,and the difference was statistically significant(P<0.05).Compared with the T2DM group,the femo-ral neck bone BMD,lumbar spine BMD and total hip BMD in the OP group were significantly decreased,and the difference was statistically significant(P<0.05).The serum CCL3 level in patients with T2DM accompa-nied by OP was negatively correlated with femoral neck bone BMD,lumbar spine BMD and total hip BMD(P<0.05),and the serum Semaphorin 3A level was positively correlated with femoral neck bone BMD,lum-bar spine BMD and total hip BMD(P<0.05).The results of multivariate Logistic regression analysis showed that CCL3 was an independent risk factor for T2DM with OP(P<0.05),and Semaphorin 3A was an inde-pendent protective factor for T2DM with OP(P<0.05).The results of ROC curve analysis showed that the area under the curve(AUC)of serum CCL3,Semaphorin 3A and their combination diagnosis of T2DM accom-panied by OP were 0.810,0.802 and 0.869,respectively.The AUC of the combined diagnosis of T2DM ac-companied by OP was superior to their individual diagnoses(Zcombination-CCL3=2.235,Zcombination-Semaphorin 3A=2.021,P=0.025,0.043).Conclusion The serum CCL3 level is increased in patients with T2DM accompanied by OP,and the Semaphorin 3A level is decreased.The combination of the two has certain diagnostic value for T2DM accompanied by OP.

Adrenal vein sampling is required for the staging diagnosis of primary aldosteronism,and the frames in which the adrenal veins are presented are called key frames.Currently,the selection of key frames relies on the doctor's visual judgement which is time-consuming and laborious.This study proposes a key frame recognition algorithm based on deep learning.Firstly,wavelet denoising and multi-scale vessel-enhanced filtering are used to preserve the morphological features of the adrenal veins.Furthermore,by incorporating the self-attention mechanism,an improved recognition model called ResNet50-SA is obtained.Compared with commonly used transfer learning,the new model achieves 97.11%in accuracy,precision,recall,F1,and AUC,which is superior to other models and can help clinicians quickly identify key frames in adrenal veins.

Objective:To observe the effect of epigallocatechin gallate (EGCG) on intestinal injury in sepsis, and to investigate the effect on endoplasmic reticulum stress (ERS) apoptotic pathway.Methods:Sixty male SD rats were selected and divided into five groups according to the randomized numeric table method: the sham operation group (Sham group), the cecal ligation and puncture sepsis group (CLP group), the sepsis+EGCG low-dose group (postoperative intraperitoneal injection of EGCG 25 mg/kg, EL group), the sepsis+EGCG medium-dose group (postoperative intraperitoneal injection of EGCG 50 mg/kg, EM group), and sepsis+EGCG high-dose group (postoperative intraperitoneal injection of EGCG 75 mg/kg, EH group), each group with 12 rats. The rats in each group were executed 24 h after modeling and specimens were collected. Inflammatory factors in serum were detected by enzyme-linked immunosorbent assay. Pathological changes of ileum were observed under light microscope after hematoxylin eosin staining and evaluated according to the Chiu's score. The intestinal tissues were stained for tight junction protein-1 (CLDN1, Claudin-1), phosphorylated protein kinase R-like endoplasmic reticulum kinase (p-PERK), protein kinase RNA-like endoplasmic reticulum kinase (PERK), cysteinyl aspartate specific protein-12 (Caspase-12), and CCAAT enhancer-binding protein homologous protein (C/EBP-homologous protein antibody, CHOP) protein expression was detected by protein immunoblotting assay. The positive areas of Claudin-1, p-PERK, CHOP, and Caspase-12 in intestinal tissue were detected by immunohistochemistry.Results:Compared with the Sham group, the serum levels of interleukin (IL)-1β, IL-6, tumor necrosis factor-α, and the Chiu's score of rats in the CLP group were increased (all P<0.05). The ileal mucosal tissues showed reduced expression of Claudin-1, ERS apoptosis-associated protein p-PERK, CHOP, and Caspase-12 expression were increased (all P<0.05). Compared with the CLP group, the intestinal injury in rats was alleviated after the administration of low, medium and high dose EGCG intervention (all P<0.05). The serum inflammatory factor level, Chiu's score and the protein expression level and positive area of ERS apoptosis-related proteins, p-PERK, CHOP, and Caspase-12 in the small intestinal tissues of EL group were further reduced compared with that of the CLP group were further decreased, and EM group was further decreased than EL group, and EH group was further decreased than EM group (all P<0.05). The protein expression level and positive area of Claudin-1 in small intestinal tissues of EL group were further increased compared with that of CLP group (both P<0.05), and EM group was further increased compared with that of EL group and EH group was further increased compared with EM group (all P<0.05). Conclusions:EGCG may have a protective effect on intestinal injury in septic rats by inhibiting the activation of ERS-induced apoptotic pathway, and the efficacy of high-dose EGCG has a better effect.

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.

Background Aedes albopictus is the dominant mosquito species in residential areas in Shanghai. There are many types of small containers with accumulated water in residential areas, providing a large number of breeding environments for Aedes alpopicuts and leading to an increasing transmission risk of mosquito-borne diseases. Objective To use random forest to predict breeding of Aedes mosquitoes in small aquatic container habitat in two concentrated reconstruction communities of rural areas in Shanghai, and to understand associated influence of environmental factors on the breeding of Aedes mosquitoes in the process of urbanization.Methods Small-scale habitat surveys of Aedes mosquitoes were carried out in two suburb concentrated reconstruction communities (Community A and B) in Shanghai, and the environment where the habitat was located was recorded and analyzed in both communities. The habitat where eggs, larvae, or pupae were found was recorded as positive. Spatial weight matrix was applied on a household basis, and global Moran's I index was used to carry out spatial autocorrelation analysis on the small-scale habitat and positive habitat in the environment of the two communities. When Moran's I is greater than 0, it means that the data present a positive spatial correlation; when Moran's I is less than 0, it means that the data are spatially negatively correlated; when Moran's I is 0, the spatial distribution is random. Combining the results of P and Z values, we explored the spatial distribution characteristics of small-scale habitat and positive habitat in the community environment. Random forest algorithm in machine learning was used to classify and sort environmental-related factors, and predict the breeding of Aedes mosquitoes in small aquatic habitat; receiver operating characteristic (ROC) curve was used to carry out model fitting evaluation. Results The environmental factors including building location (χ²=23.35, P<0.001), open space (χ²=8.83, P=0.003), and having trees (χ²=11.02, P=0.001) had a significant impact on the positive rate of small-scale habitat. The results of spatial characteristics analysis showed that the global Moran's I index of small-scale habitat was −0.092 (Z=−1.09, P=0.274) in Community A and 0.034 (Z=0.52, P=0.602) in Community B, and the global Moran's I index of positive habitat was −0.092 (Z=−1.14, P=0.255) in Community A and 0.070 (Z=0.95, P=0.342) in Community B. Since the P values of Community A and B were greater than 0.1 and the Z values were between −1.65 and 1.65, for both small-scale habitat and positive habitat the spatial characteristics were randomly distributed and no significant spatial aggregation was found. In the fitted random forest algorithm classification prediction model with the top 10 characteristic factors of importance, the area under curve (AUC) value was 0.95, and the prediction fitting effect was satisfactory. The results of classification and sorting indicated that counts of household small-scale habitat and positive habitat were the most important factors for breeding. Conclusion The random forest model constructed by environmental factor indicators can be used to predict the breeding situation of Aedes mosquitoes in small-scale aquatic habitat, and provide a basis for scientific prevention and control of mosquito breeding for the target area.

Objective:To establish a high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of voliconazole (VRC), posaconazole (PCZ), and linazolam (LNZ) in human serum.Methods:This study is a methodological validation by LC-MS/MS. The blood concentration results of VRC, PCZ, and LNZ in our hospital′s anti-infection patients were collected. Voriconazole, Posaconazole, and Linezolid were accurately weighed and prepared. Linezolid-[2H3] was used as the internal standard. After gradient elution on the ACE PFP column, the residuals were analyzed by LC-MS/MS in the positive electrospray ionization mode and multiple reaction monitor (MRM) mode. The method′s linearity, precision, lower limit of detection, and recovery rate were validated according to standard guidelines.Results:The linear correlation coefficient ( r) of the standard curve was above 0.99 ( r>0.99). The linear range of VRC and PCZ were 0.10 mg/L~10.00 mg/L, and the lower limit of detection were 0.01 mg/L. The linear range of LNZ was 0.50 mg/L~50.00 mg/L, and the lower limit of detection was 0.05 mg/L. The recoveries of VRC, PCZ and LNZ were 90.96%-103.18%, 91.84%-99.17%, and 97.04%-100.41%, respectively. Intra-and inter-batch precision (% CV) for VRC were less than 8.30%. Intra-and inter-batch precision (% CV) for PCZ was less than 9.78%. Intra-and inter-batch precision (% CV) for LNZ was less than 7.14%. Drug concentrations in 155 cases of VRC, 44 cases of PCZ, and 59 cases of LNZ were detected. Conclusion:We have established an LC-MS/MS method for the rapid, accurate, highly specific determination of VRC, PCZ, and LNZ concentrations in human serum. This method is suitable for analyzing large clinical sample sets.

OBJECTIVE: To investigate whether ferroptosis exists in sepsis induced intestinal injury, and to verify the association between ferroptosis in sepsis induced intestinal injury and intestinal inflammation and barrier function by stimulating and inhibiting the nuclear factor E2-related factor 2/glutathione peroxidase 4 (Nrf2/GPX4) pathway. METHODS: Forty-eight SPF grade male Sprague-Darvley (SD) rats with a body weight of 220-250 g were divided into sham operation group (Sham group), sepsis group (CLP group), sepsis+iron chelating agent deferoxamine (DFO) group (CLP+DFO group) and sepsis+ferroptosis inducer Erastin group (CLP+Erastin group) using a random number table method, with 12 rats in each group. The sepsis model was established by cecal ligation and puncture (CLP). The Sham group was only performed with abdominal opening and closing operations. After modeling, the CLP+DFO group received subcutaneous injection of 20 mg/kg of DFO, the CLP+Erastin group was intraperitoneally injected with 20 mg/kg of Erastin. Each group received subcutaneous injection of 50 mg/kg physiological saline for fluid resuscitation after surgery, and the survival status of the rats was observed 24 hours after surgery. At 24 hours after model establishment, 6 rats in each group were selected. First, live small intestine tissue was taken for observation of mitochondrial morphology in smooth muscle cells under transmission electron microscopy and determination of reactive oxygen species (ROS). Then, blood was collected from the abdominal aorta and euthanized. The remaining 6 rats were sacrificed after completing blood collection from the abdominal aorta, and then small intestine tissue was taken. Western blotting was used to detect the expression of intestinal injury markers such as Claudin-1 and ferroptosis related proteins GPX4 and Nrf2. Observe the pathological changes of small intestine tissue using hematoxylin-eosin (HE) staining and complete Chiu score; Detection of tumor necrosis factor-α (TNF-α), interleukins (IL-1β, IL-6) levels in serum using enzyme-linked immunosorbent assay (ELISA). The levels of serum iron ions (Fe³⁺), malondialdehyde (MDA), and D-lactate dehydrogenase (D-LDH) were measured. RESULTS: (1) Compared with the Sham group, the 24-hour survival rate of rats in the CLP group and CLP+Erastin group significantly decreased (66.7%, 50.0% vs. 100%, both P < 0.05), while there was no significant difference in the CLP+DFO group (83.3% vs. 100%, P = 0.25). (2) Western blotting results showed that compared with the Sham group, the expressions of GPX4 and Claudin-1 in the small intestine tissue of the CLP group, CLP+DFO group, and CLP+Erastin group decreased significantly, while the expression of Nrf2 increased significantly (GPX4/β-actin: 0.56±0.02, 1.03±0.01, 0.32±0.01 vs. 1.57±0.01, Claudin-1/β-actin: 0.60±0.04, 0.96±0.07, 0.41±0.01 vs. 1.40±0.01, Nrf2/β-actin: 0.88±0.02, 0.72±0.01, 1.14±0.01 vs. 0.43±0.02, all P < 0.05). Compared with the CLP group, the expressions of GPX4 and Claudin-1 were significantly increased in the CLP+DFO group, while the expression of Nrf2 was significantly reduced. In the CLP+Erastin group, the expressions of GPX4 and Claudin-1 further decreased, while the expression of Nrf2 further increased (all P < 0.05). (3) Under the light microscope, compared with the Sham group, the CLP group, CLP+DFO group, and CLP+Erastin group showed structural disorder in the small intestinal mucosa and submucosal tissue, significant infiltration of inflammatory cells, and destruction of glandular and villous structures. The Chui score was significantly higher (3.25±0.46, 2.00±0.82, 4.50±0.55 vs. 1.25±0.45, all P < 0.05). (4) Under transmission electron microscopy, compared with the Sham group, the mitochondria in the other three groups of small intestinal smooth muscle cells showed varying degrees of volume reduction, increased membrane density, and reduced or disappeared cristae. The CLP+Erastin group showed the most significant changes, while the CLP+DFO group showed only slight changes in mitochondrial morphology. (5) Compared to the Sham group, the CLP group, CLP+DFO group, and CLP+Erastin group had serum levels of TNF-α, IL-1β, IL-6, MDA, D-LDH, and ROS in small intestine tissue were significantly increased, while the serum Fe³⁺ content was significantly reduced [TNF-α (ng/L): 21.49±1.41, 17.24±1.00, 28.66±2.72 vs. 14.17±1.24; IL-1β (ng/L): 108.40±3.09, 43.19±8.75, 145.70±11.00 vs. 24.50±5.55; IL-6 (ng/L): 112.50±9.76, 45.90±6.52, 151.80±9.38 vs. 12.89±6.11; MDA (μmol/L): 5.61±0.49, 3.89±0.28, 8.56±1.17 vs. 1.86±0.41; D-LDH (kU/L): 39.39±3.22, 25.38±2.34, 53.29±10.53 vs. 10.79±0.52; ROS (fluorescence intensity): 90 712±6 436, 73 278±4 775, 110 913±9 287 vs. 54 318±2 226; Fe³⁺ (μmol/L): 22.19±1.34, 34.05±1.94, 12.99±1.08 vs. 51.74±11.07; all P < 0.05]. Compared with CLP group, the levels of TNF-α, IL-1β, IL-6, MDA, D-LDH and ROS in CLP+Erastin group were further increased, and the content of Fe³⁺ was further decreased, the CLP+DFO group was the opposite (all P < 0.05). CONCLUSIONS Ferroptosis exists in the intestinal injury of septic rats, and stimulating or inhibiting ferroptosis through the Nrf2/GPX4 pathway can effectively intervene in the inflammatory state and intestinal mechanical barrier of the body.
Rats ; Male ; Animals ; NF-E2-Related Factor 2 ; Tumor Necrosis Factor-alpha ; Ferroptosis ; Reactive Oxygen Species ; Actins ; Claudin-1 ; Interleukin-6 ; Sepsis/metabolism* ; Iron

Chronic Kidney Disease (CKD) has become one of the main diseases severally threatening human health, which has aroused global attention. Acute Kidney Injury (AKI) is the main cause of CKD. Early diagnosis of AKI is the most effective way to prevent CKD. More and more new biomarkers of renal injury will be found through genomics, proteomics, and metabolomics methodologies. The clinical application of a single marker in the real world will be subject to certain restrictions because of the etiological diversity and personal differences. Kidney damage monitoring models will be established by artificial intelligence method. However, standardizing the evaluation of its clinical application value is essential to screen out truly valuable kidney injury biomarkers or monitoring models, to promote the clinical application of kidney injury markers, and to effectively reduce the incidence of CKD.

Acute kidney injury is a common and severe clinical event, the morbidity and mortality of acute kidney injury are increasing steadily, which is related with heavy burden to the country and social economy. Early diagnosis and intervention could significantly reduce the occurrence and progression of acute kidney injury. Therefore, it is of particular importance to find sensitive, specific, and economic biomarkers for acute kidney injury. In recent years, efforts have been made on exploring proteins in blood and urine as biomarkers of acute kidney injury. Present review summarizes progress in this field based on results of recent basic and clinical research.