Patients supported by extracorporeal membrane oxygenation (ECMO) are often in a state of severe hypermetabolism and hypercatabolism, making nutritional support difficult, and there are significant deviations in estimating energy consumption by traditional formulas. On May 1, 2025, a 42-year-old male patient with acute extensive anterior myocardial infarction was admitted to the First Dongguan Affiliated Hospital of Guangdong Medical University. After admission, the patient underwent coronary stent implantation, and subsequently developed severe complications such as cardiac failure, cardiogenic shock and electrical storm, with an Acute Physiology and Chronic Health Evaluation II of 34 points. A series of invasive supportive treatments were successively administered to the patient, including tracheal intubation with mechanical ventilation, intra-aortic balloon pump (IABP), veno-arterial ECMO (VA-ECMO), continuous renal replacement therapy (CRRT) and cardiac pacemaker implantation. During the treatment, resting energy expenditure (REE) of the patient was monitored in real time by metabolic cart (indirect calorimetry), and the monitoring results were compared with those calculated by the traditional Harris-Benedict formula. Individualized nutritional support regimens were formulated based on the metabolic cart monitoring data: the ratio of enteral to parenteral nutrition was 1 : 2 during VA-ECMO treatment, and adjusted to 2 : 1 in the late post-weaning period with the addition of ω-3 polyunsaturated fatty acids. Meanwhile, an integrated traditional Chinese and Western medicine approach was adopted to regulate gastrointestinal function and repair digestive tract damage. Monitoring results showed that the REE value measured by metabolic cart decreased to one-third of the pre-VA-ECMO level during VA-ECMO treatment, which was correlated with ECMO machine parameters. Various invasive treatments exerted complex effects on the body's nutritional and metabolic status and also interfered with the monitoring indicators of the metabolic cart. Guided by metabolic cart monitoring, precise nutritional therapy was implemented in combination with multidisciplinary comprehensive treatment, which maintained the patient's metabolic stability, promoted the gradual improvement of organ functions, and laid a solid foundation for successful weaning from supportive devices and favorable prognosis. This case suggests that individualized clinical nutritional therapy under the guidance of metabolic cart is conducive to optimizing energy supply for ECMO patients and improving clinical outcomes, thus possessing important clinical application value.

Abstract In early sepsis the direction in which blood immune-cell transcriptional programmes move may carry prognostic information beyond a single baseline measurement, but whether such trajectory associations survive independent testing is unknown. We scored five immune modules, frozen before analysis, in three public longitudinal whole-blood microarray sepsis cohorts and fitted a logistic model ladder fixed in advance to the change per 24 hours in the two cohorts with mortality data (82 patients, 24 deaths), pooling by inverse-variance fixed-effect meta-analysis with Benjamini-Hochberg control. No association survived correction for multiple testing. The two leading signals were a rising CD4/NK lymphocyte trajectory associated with lower mortality (pooled odds ratio 0.53, 95% confidence interval 0.31 to 0.90) and a rising emergency-granulopoiesis trajectory associated with higher mortality (1.60, 0.92 to 2.79), both per one standard deviation. We then tested both in an independent transcriptomic cohort with serial sampling (63 patients, 15 deaths), scored by the identical frozen method, and against their cell-count analogues in an intensive-care database of 12,607 adults meeting Sepsis-3 criteria, of whom 744 to 4,206 had the serial measurements each analogue required. Independent testing separated the two signals, in the order opposite to the one discovery had suggested. The emergency-granulopoiesis association was reproduced in direction and effect size without reaching conventional significance on its own (validation odds ratio 1.72, 0.92 to 3.20, p=0.088; pooled 1.65, 1.09 to 2.50), was positive in all nine sensitivity analyses, each fixed before the estimates were examined, and was supported by two of its three analogues, including the neutrophil-to-lymphocyte ratio (1.31, 1.20 to 1.42). The CD4/NK association did not reproduce (1.06, 0.59 to 1.89), was null in the window most favourable to it, and received no support from an analogue well powered to detect the discovery effect. The discovery signal that looked most consistent failed independent testing.

Background Limited data are available regarding disseminated intravascular coagulation (DIC) scores after liver transplantation (LT). As Chinese DIC Scoring System (CDSS) is widely accepted for assessing coagulation in China, this study was aimed to investigate the prognostic value of CDSS scores in patients with undergoing LT. Method A retrospective cohort study was conducted on patients who underwent LT from November 2009 to October 2021. We validated CDSS criteria by comparing with International Society on Thrombosis and Hemostasis (ISTH) score. Additionally, its prognostic value was evaluated with receiver operating characteristic (ROC) curves and odds ratio based on mortality rates at 28, 60, and 90 days, as well as the correlations between the CDSS score and acute physiological and chronic health assessment II (APACHE II), sequential organ failure assessment (SOFA) scores at 90-day mortality. Results A total of 569 LT patients were enrolled, of which 80 patients developed DIC with CDSS score and 305 patients with ISTH score. Patients with DIC using the CDSS exhibited higher APACHE II and SOFA scores than those with ISTH score. The incidences of acute kidney injury, infection, lymphocytopenia and mortality were higher in DIC patients with CDSS than in those with ISTH. When assessing the prognostic value for 28-day mortality, the CDSS demonstrated higher sensitivity (64.61% vs. 50.77%), but lower specificity (73.62% vs. 88.89%) compared to the ISTH, the areas under ROC (AUC) for the CDSS and ISTH scores were 0.739, 0.741 (p < 0.05) and the odds ratios (OR) for the CDSS and the ISTH were 6.228, 3.597, respectively (p < 0.05). The ORs for predicting mortality with 60-day (7.719 vs. 3.95) and 90-day (7.582 vs. 3.95) criteria with CDSS were higher than those with ISTH (p < 0.05). The Spearman's rank correlation coefficients between the CDSS and APACHE II scores, and the SOFA scores were 0.217 and 0.422, respectively, compared to 0.19 and 0.371 for the ISTH score (p < 0.001). Conclusion Disseminated intravascular coagulation presents a life-threatening complication in perioperative period of LT. The CDSS score has better prognostic value than the ISTH score for DIC patients after LT. A prospective randomized controlled study should be designed to further evaluate the findings.

Background:The high mortality rate of severe heat stroke is mainly related to multiple organ dysfunction syndrome (MODS), and respiratory failure caused by acute lung injury (ALI) is a significant factor in the development of MODS during the course of severe heat stroke. Previous research has demonstrated that severe heat stroke-induced acute lung injury (sHS-ALI) is associated with an increase in reactive oxygen species (ROS) in vascular endothelial cells (VECs), but the specific initiating factors and intermediate mechanisms involved are unclear. Methods:In this study, the mRNA profiles of mouse lung tissues were analysed using high-throughput sequencing. Genome-wide knockout was performed using CRISPR-Cas9 technology to identify a cohort of differentially expressed genes that promote human umbilical vein endothelial cells survival after heat stress. The expression of key proteins [fibroblast growth factor 23 (FGF23), phosphorylated fibroblast growth factor receptor-1 (p-FGFR-1), FGFR-1, phosphorylated phospholipase C-γ2 (p-PLC-γ2), PLC-γ2, p-p47phox, p67phox, p22phox, p40phox, and nicotinamide adenine dinucleotide phosphate oxidase isoform 2 (NOX2)] involved in the FGF23/FGFR-1 mechanism was examined using western blotting and immunohistochemistry. Results:In this study, we first screened sHS-ALI target genes by cross-comparison in vivo and in vitro and found that FGF23 is the upstream promoter of sHS-ALI. Subsequent investigations involving the interference or inhibition of FGF23 expression revealed that FGF23 induced FGFR-1 Y766 phosphorylation during heat stress-induced VECs damage. In addition, FGF23 participated in NOX2 activation and ROS accumulation and was involved in the process of sHS-ALI. These findings indicated that the FGFR-1 Y766 site mutation strongly suppressed the production of p-PLC-γ2 and heat stress-induced NOX2-ROS activation in VECs. More importantly, mutation of the FGFR-1 Y766 phosphorylation site had no effect on FGF23 expression, and it was impossible to significantly induce the expression of p-PLC-γ2. Moreover, NOX2-ROS activation was inhibited, even in the presence of heat stress, the recombinant FGF23 protein, or combined stimulation. Conclusions:This study confirmed that FGF23/FGFR1 signalling, as an upstream priming factor, mediated NOX2-ROS activation in VECs after heat stress, thus participating in the sHS-ALI process. FGFR-1 Y766 phosphorylation is essential for FGF23/FGFR-1 signalling activation in VECs, which is involved in sHS-ALI. These findings further clarify the mechanism underlying sHS-ALI and contribute to reducing the mortality and morbidity of severe heat stroke.

Evidence for the effectiveness of the Sonoclot analyzer in improving clinical outcomes in disseminated intravascular coagulation (DIC) is lacking. To evaluate the effectiveness of an algorithm based on the Sonoclot analyzer in improving the short-term prognosis of patients with DIC. A total of 279 patients with overt DIC who were admitted to the hospital within 18 months before and after implementation of the new Sonoclot-based algorithm were enrolled in the study. They were assigned to either a conventional coagulation assay (CCA) group (n = 148) or a Sonoclot group (n = 131). Data associated with anti-coagulation and transfusion were collected and analyzed. The 30-day survival rate after hospital admission was compared between groups. The Sonoclot group had a higher 30-day survival rate than the CCA group (78.45% vs 63.64%; P = .02). The heparin dose, anticoagulation course, and major bleeding rate were significantly reduced in the Sonoclot group compared with the CCA group for all patients and for patients undergoing continuous renal replacement therapy (all P < .001). Fresh frozen plasma, platelet, and cryoprecipitate requirements were substantially lower in the Sonoclot group than in the CCA group (P = .007, .03, and .02, respectively). In a stratified analysis, improved survival rate was seen mainly in patients with moderately severe sepsis and heatstroke, with an Acute Physiology and Chronic Health Evaluation II score of 20 to 29. The Sonoclot analyzer may be useful to guide coagulation management in patients with DIC. Use of the Sonoclot-based algorithm may improve outcomes for DIC patients with moderately severe sepsis or heatstroke.

BACKGROUND:Thrombocytopaenia is a known predictor of poor outcomes in trauma and sepsis patients, its prognostic role in exertional heatstroke (EHS) remains underexplored. This study aimed to evaluate the association between persistent thrombocytopaenia and mortality in critically ill EHS patients. METHODS:In this retrospective cohort study, we analyzed data from 217 EHS patients admitted to the intensive care unit (ICU) between November 2009 and October 2023. Demographic, biochemical (baseline, 24 h, and 48 h), and inflammatory marker data were collected. Persistent thrombocytopaenia was defined as a platelet count <100 × 109/L at both 24 h and 48 h postadmission. Multivariate logistic regression identified predictors of 28-day mortality, and Kaplan-Meier survival curves with log-rank tests were used to compare outcomes between groups. RESULTS:Among the 217 patients, 19 (8.8%) patients died within 28 days. Nonsurvivors presented significantly worse liver/kidney function, coagulopathy (DIC incidence: 89.5% vs. 26.8%, p < 0.001), and AKI incidence (100% vs. 38.4%, p < 0.01). The platelet counts in nonsurvivors were markedly lower than those in survivors at admission (71 vs. 166 × 109/L), 24 h (49 vs. 134 × 109/L), and 48 h (56 vs. 131 × 109/L) (all p < 0.001). Persistent thrombocytopaenia (63 patients, 29.0%) independently predicted mortality (adjusted OR = 17.44, 95% CI: 4.76-63.86; p < 0.001) and was correlated with higher DIC (84.1% vs. 11.0%, p < 0.001) and AKI (63.5% vs. 35.7%, p < 0.001) rates. Survival analysis confirmed significantly increased 28-day mortality in thrombocytopaenia patients (25.4% vs. 1.9%, p < 0.001). CONCLUSION:Persistent thrombocytopaenia within 48 h of ICU admission is a robust predictor of mortality and multiorgan dysfunction in patients with EHS.

Abstract Background: Prolonged exposure to high temperatures and high humidity can cause a number of harmful stress reactions, including severe heat stroke and multiple organ dysfunction syndrome (MODS), which constitute a direct threat to people's lives. The pathophysiological response to severe heat stroke is more likely to be due to systemic inflammatory response syndrome (SIRS) secondary to heat injury. Consequently, in order to find more effective preventative and therapeutic strategies, it is worthwhile to begin with a research of the mechanism of severe heat stroke. Applying single-cell RNA sequencing (scRNA-seq) technology to further understand the biological functions of various cell types in severe heat stroke will facilitate the intervention of severe heat stroke. Methods: In this experiment, mice severe heat stroke model were constructed and peritoneal lavage fluid was collected. Single-cell RNA sequencing (scRNA-seq) was used, combined with cluster analysis, Uniform Manifold Approximation and Projection (UMAP) visualization, and other bioinformatics methods to distinguish cell subgroups while using gene set variation analysis (GSVA), gene ontology (GO) function enrichment and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment to reveal the biological function of each cell subgroup. Results: This study identified 17-cell clusters with cell type-specific gene expression patterns and cell subpopulations in the intraperitoneal lavage fluid, and these 17-cell clusters were divided into 5 major cell lineages: T-NK cells (clusters 6,7,11,17), myeloid cells (clusters 1,5,8,10,12,14), B cells (clusters 2,3,4,9,15), mast cells (clusters 13), erythroid cells (clusters 16). Simultaneously, the data demonstrated that, in comparison to other cell lineages, the rise in myeloid cells was the most notable; The subpopulation of myeloid cells is primarily composed of two cell types and twelve cell clusters: macrophages and dendritic cells (DC). The macrophages comprise clusters 1, 2, 3, 5, 6, 7, 9, 10, and the DCs include clusters 8, 11, and 12. Meanwhile given the presence of a majority of macrophages in the myeloid cells subpopulation, the number of cells in each cell cluster of of macrophages was analyzed , and it was found that the number of cells in clusters 1,3,8,10 significantly decreased, while the number of cluster 2,4,5,6 cells significantly increased after severe heat stroke. Additionally, the top 20 differentially expressed genes (DEGs) in recruited and resident macrophages were screened, and the top 20 DEGs with substantial changes were subjected to KEGG and GO analyses. Furthermore, cluster7 was a newly emerging a cell subpopulation after severe heat stroke, then GO and KEGG analyses were then conducted on the top 200 DEGs that showed significant alterations. These findings suggest that distinct cell subpopulations are impacted by biological processes and signaling pathways. Conclusions: ScRNA-seq analysis of the intraperitoneal lavage fluid from mice suffering from severe heat stroke demonstrated the diversity of cell differentiation and the distinct characteristics of cell subpopulations, offering new insights into the pathogenic mechanism and the development of potential treatments for severe heat stroke.

While serum 25-hydroxyvitamin D (25[OH]D) deficiency is prevalent in chronic kidney disease (CKD), the effects of 25(OH)D deficiency on cardiovascular mortality and kidney outcomes in patients with early-stage CKD remain incompletely understood. This multicenter retrospective cohort study included adult patients with stages 1–3 CKD from 19 medical centers across China between January 2000 and May 2021. The primary outcome was cardiovascular mortality. The secondary study outcome included CKD progression (defined as a sustained > 40

Exosomes are 30–150 nm small extracellular vesicles (sEVs) which are highly stable and encapsulated by a phospholipid bilayer. Exosomes contain proteins, lipids, RNAs (mRNAs, microRNAs/miRNAs, long non-coding RNAs/lncRNAs), and DNA of their parent cell. In pathological conditions, the composition of exosomes is altered, making exosomes a potential source of biomarkers for disease diagnosis. Exosomes can cross the blood–brain barrier (BBB), which is an advantage for using exosomes in the diagnosis of central nervous system (CNS) diseases. Neuropsychiatric diseases belong to the CNS diseases, and many potential diagnostic markers have been identified for neuropsychiatric diseases. Here, we review the potential diagnostic markers of exosomes in neuropsychiatric diseases and discuss the potential application of exosomal biomarkers in the early and accurate diagnosis of these diseases. Additionally, we outline the limitations and future directions of exosomes in the diagnosis of neuropsychiatric diseases.

Background:There is much debate on the optimal treatment approach of papillary thyroid carcinoma (PTC). Different guidelines base recommendations on various risk factors. While diagnosing the various risk factors is difficult due to the technical limitations, intraoperative frozen section (IFS) may be a feasible method. We aim to real-time evaluate the multiple risk factors, including lymph node metastasis (LNM), extrathyroidal extension (ETE), multifocality using IFS, and then identify a more effective surgical plan, which may help avoid the need for a second surgery and improve prognosis of patients. Methods:We retrospectively reviewed the medical records of 364 patients from January 1, 2021 to December 31, 2021. All the patients were initially recommended to undergo a hemithyroidectomy (HT) with isthmusectomy and ipsilateral central compartment neck dissection (CCND). IFS would be executed immediately. Further total thyroidectomies (TTs) would be performed if: (I) results of IFS showed >5 LNM, or (II) there are 1≤ LNM ≤5 but with ETE and/or multifocal carcinoma. The patients were divided and investigated according to the extent of surgery. Results:Based on the results of IFS, 72 patients underwent TT. The TT group displayed larger average tumor diameter, greater age, higher average body mass index (BMI), and elevated incidence of hypertension and hyperlipidemia compared to the HT group. The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of IFS were 77.61%, 100%, 100%, and 88.46%, respectively. Conclusions:IFS is a highly reliable procedure. Comprehensively evaluating central compartment LNM, ETE, and multifocal carcinoma through IFS helps identify a more reasonable surgical option under the current clinical consensus, which may thus help avoid the need for a second surgery.

Purpose To explore the key genes and molecular pathways in the progression of thyroid papillary carcinoma (PTC) promoted by testosterone using RNA-sequencing technology, and to provide new drug targets for improving the therapeutic effect of PTC. Methods Orchiectomy (ORX) was carried out to construct ORX mouse models. TPC-1 cells were subcutaneously injected for PTC formation in mice, and the tumor tissues were collected for RNA-seq. The key genes were screened by bioinformatics technology. Tnnt1 expression in PTC cells was knocked down or overexpressed by transfection. Cell counting kit-8 (CCK-8), colony formation assay, scratch assay and transwell assay were adopted, respectively, for the detection of cell proliferation, colony formation, migration and invasion. Besides, quantification real-time polymerase chain reaction (qRT-PCR) and western blot were utilized to determine the mRNA and protein expression levels of genes in tissues or cells. Results Both estradiol and testosterone promoted the growth of PTC xenografts. The key gene Tnnt1 was screened and obtained by bioinformatics technology. Functional analysis revealed that overexpression of Tnnt1 could markedly promote the proliferation, colony formation, migration, invasion, and epithelial-to-mesenchymal transition (EMT) process of PTC cells, as well as could activate p38/JNK pathway. In addition, si-Tnt1 was able to inhibit the cancer-promoting effect of testosterone. Conclusion Based on the outcomes of bioinformatics and basic experiments, it is found that testosterone can promote malignant behaviors such as growth, migration, invasion and EMT process of PTC by up-regulating Tnnt1 expression. In addition, the function of testosterone may be achieved by activating p38/JNK signaling pathway.

Objective To investigate the early diagnosis value of neutrophil-side-fluorescence intensity（NE-SFI） on heat stroke（HS）-related disseminated intravascular coagulation（DIC）. Methods According to the International Society of Thrombosis and Haemostasis（ISTH） scoring criteria, thirty-four HS patients admitted to the General Hospital of Southern Theater Command from January 1, 2017 to December 31, 2018 were selected and divided into HS without DIC group（DIC score ＜5 points, n=23）and HS with DIC group（DIC score ≥5 points, n=11）. The patient’s general information, NE-SFI, and neutrophil extracellular traps（NETs）-related markers such as, dsDNA（double-stranded DNA）, myeloperoxidase（MPO） and citrullinated histone（CitH3）were compared between the two groups. Receiver operating characteristic（ROC） curve was used to analyze the early diagnostic value of NE-SFI in HS with DIC. Results There was no significant difference in age, maximum body temperature, white blood cell count and neutrophil count between the two groups（P＞0.05）. The proportion of patients in the HS with DIC group whose core temperature dropped below 38.5 ℃ within 3 hours and the GCS（Glasgow Coma Scale） score were lower than those in the HS without DIC group, while the alanine aminotransferase（ALT）, creatinine, ISTH（International Society of Thrombosis and Haemostasis） score, and the proportion of concurrent multiple organ dysfunction syndrome（MODS） in the HS with DIC group were higher than those in the HS without DIC group（P＜0.05）. On the 1st to 3rd day of onset, the NE-SFI values of the HS with DIC group were higher than those of the HS without DIC group（P＜0.001）. Compared with the serum dsDNA, MPO, and CitH3 in HS without DIC group [respectively（30.14±7.01） ng/ml,（56.39±34.64） pg/ml,（320.26±89.60） ng/μl], the serum dsDNA, MPO and CitH3 levels in HS with DIC group [respectively（372.93±135.77） ng/ml,（108.32±38.58） pg/ml,（600.18±183.74） ng/μl]are significantly increased（P＜0.001）. Spearman correlation analysis showed that the value of NE-SFI on day 1-3 were positively correlated with the levels of dsDNA, MPO and CitH3 on day 1（P＜0.05 or P＜0.01）. ROC curve analysis showed that NE-SFI on day 2 had a high value for the early diagnosis of HS complicated with DIC, and its AUC was 0.921（95%CI 0.820-1.000, P＜0.001）.Conclusion NE-SFI can be used as an effective indicator for the early diagnosis in HS complicated with DIC.

Introduction Damage to endothelial glycocalyx (EGCX) can lead to coagulation disorders in sepsis. Heat stroke (HS) resembles sepsis in many aspects; however, it is unclear whether EGCX injury is involved in its pathophysiology. The purpose of this study was to examine the relationship between the damage of EGCX and the development of coagulation disorders during HS. Methods We retrospectively collected 159 HS patients and analyzed coagulation characteristics and prognosis of HS patients with or without disseminated intravascular coagulation (DIC). We also replicated a rat HS model and measured coagulation indexes, pulmonary capillary EGCX injury in HS rats. Finally, we evaluated the effect of the antioxidant N-acetylcysteine (NAC) on HS-initiated EGCX injury and coagulation disorders. Results Clinical data showed that HS patients complicated with DIC had a higher risk of death than HS patients without DIC. In a rat HS model, we found that rats subjected to heat stress developed hypercoagulability and platelet activation at the core body temperature of 43°C, just before the onset of HS. At 24 h of HS, the rats showed a consumptive hypo-coagulation state. The pulmonary capillary EGCX started to shed at 0 h of HS and became more severe at 24 h of HS. Importantly, pretreatment with NAC substantially alleviated EGCX damage and reversed the hypo-coagulation state in HS rats. Mechanically, HS initiated reactive oxidative species (ROS) generation, while ROS could directly cause EGCX damage. Critically, NAC protected against EGCX injury by attenuating ROS production in heat-stressed or hydrogen peroxide (H 2 O 2 )-stimulated endothelial cells. Discussion Our results indicate that the poor prognosis of HS patients correlates with severe coagulation disorders, coagulation abnormalities in HS rats are associated with the damage of EGCX, and NAC improves HS-induced coagulopathy, probably through its protection against EGCX injury by preventing ROS generation.

Aim: To investigate the mechanism of p53-mediated suppression of heat stress-induced oxidative stress damage by manganese superoxide dismutase (MnSOD) in endothelial cells (ECs). Methods: Primary ECs isolated from mouse aortas were used to examine the effects of heat stress on vascular ECs viability and apoptosis. We measured MnSOD expression, reactive oxygen species (ROS) production, p53 expression, viability, and apoptosis of heat stress-induced ECs. We also tested the protective effects of MitoQ10, a mitochondrial-targeted antioxidant, and Pifithrin-α, a p53 inhibitor, in ECs from a mouse model of heat stroke. Results: Heat stress increased cellular apoptosis, ROS production, and p53 expression, while reducing cellular viability and MnSOD expression in ECs. We also showed that the suppression of MnSOD expression by heat stress in ECs was mediated by interactions between p53 and Sp1. Furthermore, MitoQ10 and Pifithrin-α alleviated heat stress-induced oxidative stress and apoptosis in ECs. Conclusion: Our results revealed that p53-mediated MnSOD downregulation is a key mechanism for heat stress-induced oxidative stress damage in ECs and indicated that MitoQ10 and Pifithrin-α could be potential therapeutic agents for heat stroke.

BACKGROUND The mechanisms underlying heat stroke (HS)-induced hippocampal injury remain unclear. This study aimed to evaluate the HS-induced metabonomics of hippocampal and cerebellar transmitters. METHODS The HS model was established with male Sprague-Dawley rats subjected to heat exposure of up to 42 °C at a humidity of (55.0±5.0)%. The hippocampal and cerebellar transmitters and metabolites of rats were tested via ultra-high-performance liquid chromatography-mass spectrometry (UPLC-MS/MS). The primary transmitters and metabolites were identified by principal component analysis (PCA) and orthogonal partial least square-discriminant analysis (OPLS-DA). The major metabolic pathways for HS were selected after enrichment. The brain injury was evaluated by histological tests. RESULTS HS induced hippocampal and cerebellar injuries in rats. HS upregulated the protein levels of hippocampal glutamate, glutamine, gamma-aminobutyric acid, L-tryptophan (Trp), 5-hydroxy-indoleacetic acid, and kynurenine; however, it downregulated asparagine, tryptamine, 5-hydroxytryptophan, melatonin, 3,4-dihydroxyphenylalanine (L-DOPA), and vanillylmandelic acid. HS also sharply elevated the protein levels of cerebellar methionine and Trp, and decreased the levels of serotonin, L-alanine, L-asparagine, L-aspartate, cysteine, norepinephrine, spermine, spermidine, and tyrosine. Hippocampal glutamate, monoamine transmitters, cerebellar aspartate acid, and catecholamine transmitters' metabolic pathways were identified as the main metablic pathways in HS. CONCLUSION The hippocampus and cerebellum were injured in rats with HS, possibly induced the disorder of hippocampal glutamate and serotonin metabolism, cerebellar aspartate acid and catecholamine transmitter metabolism, and related metabolic pathways.

Background: Heatstroke is a life-threatening disease. Present study was aimed to investigate the mechanism in heat induced intestinal epithelial cell death.Method: Heat stress in vitro model was established on IEC cells with 42 & DEG;C for 2 h. Caspase-8 inhibitor, Caspase-3 inhibitor, RIP3 inhibitor, TLR3 agonist, poly(I:C) and p53 knockdown were used to determine the signaling pathway. Heatstroke in vivo model was established on C57BL/6 mice, with a temperature of 35.5 & DEG;C & PLUSMN;0.5 & DEG;C and a relative humidity of 60% & PLUSMN; 5%. The intestine necroptosis and inflammatory cytokines were measured. Pifithrin & alpha; (3 mg/kg) and p53 knockout mice were used to evaluate the role of p53.Results: Heat stress-induced reduction of cell viability was remarkable reversed by RIP3 inhibitor. Heat stress induced upregulation of TLR3 and facilitate the formation of TRIF-RIP3 complex. The heat stress induced upregulation of RIP3 and p-RIP3 were normalized by the deletion of p53. Meanwhile, p53 knockout decreased TLR3 expression and blocked the formation of TLR3-TRIF complex. The deletion of p53 blocked the decreased cell viability and restored the activation of RIP3-MLKL signaling after heat stress, however, which were abolished by re-expression of p53 via Tp53 OE. Increased the expression of TLR3 in the p53-deficient cells could not affect the heat stress induced necrotic cell death, which suggests that heat stress induced necroptosis via TLR3-TRIFRIP3 signaling pathway is dependent on p53.Conclusion: Heat stress promoted p53 phosphorylation, then upregulated TLR3 and enhanced the interaction of TRIF-RIP3, which would activate the RIP3-MLKL signaling pathway to mediate necroptosis in intestinal epithelial cells.

PurposeThyroid cell lines are useful tools to study the physiology and pathology of the thyroid, however, they do not produce or secrete hormones in vitro. On the other hand, the detection of endogenous thyroid hormones in primary thyrocytes was often hindered by the dedifferentiation of thyrocytes ex vivo and the presence of large amounts of exogenous hormones in the culture medium. This study aimed to create a culture system that could maintain the function of thyrocytes to produce and secrete thyroid hormones in vitro.MethodsWe established a Transwell culture system of primary human thyrocytes. Thyrocytes were seeded on a porous membrane in the inner chamber of the Transwell with top and bottom surfaces exposed to different culture components, mimicking the 'lumen-capillary' structure of the thyroid follicle. Moreover, to eliminate exogenous thyroid hormones from the culture medium, two alternatives were tried: a culture recipe using hormone-reduced serum and a serum-free culture recipe.ResultsThe results showed that primary human thyrocytes expressed thyroid-specific genes at higher levels in the Transwell system than in the monolayer culture. Hormones were detected in the Transwell system even in the absence of serum. The age of the donor was negatively related to the hormone production of thyrocytes in vitro. Intriguingly, primary human thyrocytes cultured without serum secreted higher levels of free triiodothyronine (FT3) than free thyroxine (FT4).ConclusionThis study confirmed that primary human thyrocytes could maintain the function of hormone production and secretion in the Transwell system, thus providing a useful tool to study thyroid function in vitro.

Objective To explore the the effect and possible mechanism of heat stress at different rewarming time on the damage of human skeletal muscle cells(HSKMC).Methods HSKMC in the logarithmic growth phase were divided into the heat stress group and control group.Cells in the control group were cultured normally,and the heat stress models were established in the heat stress group and then were rewarmed in 37℃and 5%CO2 cell incubator for 0,6,12 and 24 h,respectively.The cell survival rate was detected by CCK-8,the cell ultrastructure was observed by transmission elec-tron microscopy,TRPV4 protein was detected by Western blotting,and TRPV4 mRNA was detected by real-time fluores-cence quantitative PCR.HSKMC in the logarithmic growth phase were randomly divided into the HS group,GSK+HS group,HC+HS group,and NC group.Cells in the NC group were cultured normally,and the heat stress models were es-tablished in the other three groups,and the rewarming time was 12 h.Cells in the GSK+HS group and HC+HS group were given TRPV4 agonist GSK1016790A(500 nmol)and TRPV4 inhibitor HC-067047(5 μmol)at 0.5 h before heat stress,and Ca2+ level was detected by flow cytometry.Results Compared with the control group,the survival rate of cells in heat stress group decreased after rewarming for 0,6,12 and 24 h(P＜0.05).With the extension of rewarming time,the survival rate of cells in the heat stress group decreased successively,and significant difference was found between these two groups(all P＜0.05).Compared with the control group,ultrastructural changes were observed in the cells in the heat stress group after rewarming for 0 and 6 h,which were mainly manifested as cell swelling,enlarged volume,cytoplasmic vacuolation,obvious mitochondrial swelling,degranulation,double ridge loss,widening of perinuclear space,nucleus condensation and contraction,and interruption of some envelope continuity.After rewarming for 12 and 24 h,the cells in the heat stress group even showed nucleolysis and disappearance,nucleus fragmentation,chromatin boundary aggrega-tion,and cytoplasmic structure disintegration and granulation.Compared with the control group,the relative expression levels of TRPV4 mRNA and protein in heat stress group increased after rewarming for 0,6,12 and 24 h(all P＜0.05).The levels of intracellular Ca2+ in the NC group,HC+HS group,HS group and GSK+HS group increased successively,and significant difference was found between groups(all P＜0.05).Conclusions Heat stress can reduce HSKMC survival rate and damage HSKMC structure.The longer rewarming time after heat stress,the greater the damage to HSKMC cells.The mechanism may be related to the promotion of TRPV4 expression and its mediated Ca2+ influx.

Erythrophagocytosis by leukemic blasts is an extremely uncommon phenomenoninacutemyeloidleukemia(AML),withlessthan1%inone previous study. 1 This finding is typically associated with myelomono-cytic or monocytic morphology (also designated as FAB-AML M4 or M5). 2 The most common chromosomal anomalies include t(8;16) or its variants, inv(8)(p11q13) and t(16;21)(p11;q22), 3–5 and, less frequently, in B-lymphoblastic leukemia (B-ALL) with ETV6-RUNX1 or BCR-ABL1 fusions. 6,7 In addition, abundant cytoplasmic granules can occasionally occur in B-ALL, but are rarely seen in T-lymphoblastic leukemia (T-ALL). Here, we describe a child diagnosed with de novo T-ALL with SIL-TAL1 fusion gene, with concurrence of significant

Emerged evidence has indicated that immunosuppression is involved in the occurrence and development of sepsis. To provide clinical practice recommendations on the immune function in sepsis, an expert consensus focusing on the monitoring and treatment of sepsis-induced immunosuppression was developed. Literature related to the immune monitoring and treatment of sepsis were retrieved from PubMed, Web of Science, and Chinese National Knowledge Infrastructure to design items and expert opinions were collected through an online questionnaire. Then, the Delphi method was used to form consensus opinions, and RAND appropriateness method was developed to provide consistency evaluation and recommendation levels for consensus opinions. This consensus achieved satisfactory results through two rounds of questionnaire survey, with 2 statements rated as perfect consistency, 13 as very good consistency, and 9 as good consistency. After summarizing the results, a total of 14 strong recommended opinions, 8 weak recommended opinions and 2 non-recommended opinions were produced. Finally, a face-to-face discussion of the consensus opinions was performed through an online meeting, and all judges unanimously agreed on the content of this consensus. In summary, this expert consensus provides a preliminary guidance for the monitoring and treatment of immunosuppression in patients with sepsis.