The root of Aconitum kusnezoffii (Caowu in Chinese, CW) is not only commonly used as a traditional Chinese medicine (TCM), but also served as a tonic in China. Due to its high toxicity, clinical poisoning cases induced by CW have frequently been reported. However, the mechanism is still unclear. In this study, Aconitum alkaloids and altered endogenous metabolites in CW poisoning patients were investigated to elucidate the possible intoxication mechanism. Eighteen alkaloids, including 6 toxic diester diterpenoid alkaloids (DDAs), were determined from the sera of patients. At the same time, 5-(diisopropylamino)amylamine (DIAAA) derivatization-ultrahigh performance liquid chromatography- quadrupole-time of flight mass spectrometry (UHPLC-Q-TOF/MS) approach was applied in the metabolomics analysis to find much more carboxyl-containing metabolites (CCMs), which are the essential components for life and critical to elucidate the mechanism of toxicity. As a result, 32 altered metabolites after poisoning were identified. Among them, hydroxyeicosatetraenoic acids (HETEs) and some dicarboxylic acids were first found to be related to Aconitum alkaloids toxicity. Finally, biological pathway analysis indicated that the significantly changed metabolites were primarily involved in amino acid metabolism, TCA cycle, fatty acid metabolism, pyruvate metabolism, arachidonic acid metabolism, sphingolipid metabolism and so on. These results can not only provide more information on the mechanism of CW intoxication but also help the clinical diagnosis of CW poisoning.

Metagenomic next-generation sequencing (mNGS) is a new high-throughput sequencing method that may have great importance in early diagnosis and clinical management of sepsis. This study aimed to detect the difference between mNGS and comprehensive routine microbiological test (CMT), and to explore the diagnostic efficacy of mNGS in septic patients. This study retrospectively analyzed 150 sepsis patients who were admitted to the intensive care units of 4 hospitals in Southwest China from October 1, 2018, to October 1, 2021, and underwent both blood mNGS and CMT. The demographic and clinical characteristics of the patients were recorded, and the distribution of pathogens was analyzed. Additionally, the diagnostic performance and concordance between mNGS and CMT were compared to evaluate the etiological diagnostic value of mNGS in sepsis patients. In this study of 150 sepsis patients, bacterial infections were identified in 126 (84.0%), viral in 15 (10.0%), and fungal in 9 (6.0%). Among the sample types, sputum was most common, representing 62% of the total cases. Bronchoalveolar lavage fluid constituted 58.7%, blood 56.0%, with other specimens including pleural fluid at 29.3%, pus at 19.3%, swabs at 9.3%, cerebrospinal fluid at 8.7%, tissue at 6.0%, and bone marrow at 5.3%. mNGS demonstrated a diagnostic accuracy of 56.0% for sepsis, with a sensitivity of 84.4%, specificity of 26.0%, a positive predictive value of 54.6%, a negative predictive value of 61.3%. Metagenomic testing enables the rapid and early identification of infectious pathogens in sepsis patients, especially fungi and viruses. The study found that mNGS has high sensitivity in diagnosing sepsis patients, particularly for fungal and viral infections. mNGS technology is beneficial for critically ill sepsis patients.

Background: Traumatic brain injury (TBI) is an important cause of disability and death worldwide. The development of neuroinflammation after TBI is related to the brain parenchyma. M1‐type microglia play important roles in this process, but the specific mechanism through which regulating microglia M1 polarization is still not fully understood. This study aimed to investigate the role of ephrin receptor A4 (EPHA4) in the M1 polarization of microglia after TBI. Methods: A TBI rat model was established by the controlled cortical impact (CCI) method, and M1 polarization of GMI‐R1 cells was induced by lipopolysaccharide (LPS) treatment. Target genes associated with the progression of TBI were screened by transcriptome sequencing; the expression of key genes and proteins was detected by real‐time quantitative PCR (RT‒qPCR), Western blot, ELISA, and immunofluorescence, and the damage of the rat brain tissue and the blood–brain barrier (BBB) was evaluated by hematoxylin‒eosin (HE) staining and Evans blue staining. Results: This study revealed that EPHA4 expression is upregulated in the brain tissue of TBI rats and that treatment with its inhibitor, KYL peptide, can improve the progression of TBI. KYL peptide intervention downregulated the levels of the proinflammatory cytokines and upregulated the levels of the anti‐inflammatory cytokines and inhibited the M1 polarization of microglia. The levels of the endoplasmic reticulum stress (ERS)‐related proteins and Ca ²⁺ were upregulated in TBI rats and downregulated after KYL peptide treatment but subsequently increased after 123C4 treatment. Our results showed that EPHA4 promoted the M1 polarization of microglia by enhancing ERS. Notably, mitogen‐activated protein kinase (MAPK) signaling was significantly enriched in TBI. In vitro studies revealed that LPS treatment promoted the activation of MAPK signaling in GMI‐R1 cells. A mechanistic study revealed that EPHA4 may activate ERS by activating the MAPK signaling pathway and promote M1 polarization of microglia after TBI. Conclusion: Our study revealed that the EPHA4/MAPK axis may be a key regulatory factor in controlling microglial M1 polarization during brain injury. Blocking this signaling axis may represent a potential therapeutic approach for improving TBI.

Tree shrews are most closely related to the primates and so possess a number of advantages in experimental studies; they have been used as an animal model in bacterial and virus infection, cancer, endocrine system disease, and certain nervous system diseases. Their olfactory ensheathing cells (OECs) are able to release several cytokines to promote neuronal survival, regeneration and remyelination. The present study used western blot analysis to identify antibody specificity in protein extracts from whole tree shrew brains to identify the specificity of p75 nerve growth factor receptor (NGFR) derived from rabbits (75 kDa). OECs were cultured and isolated, then stained and identified using the antibodies for p75NGFR. To investigate the capacity of OECs to express cytokines and growth factors, microarray technology was used, and the analysis revealed that OECs were able to express 9,821 genes. Of these genes, 44 genes were from the neurotrophic factor family, which may indicate their potential in transplantation in vivo. The present study considered the function of OECs as revealed by other studies, and may contribute to future research.

The tree shrew (Tupaia belangeri) has become an ideal model for studying various neurological diseases. However, there is no stable tree shrew model of traumatic brain injury (TBI) at present. This study aimed to establish a tree shrew TBI model and clarify the expression characteristics of inflammatory factors after brain injury, as well as the polarization characteristics of glial cells. Twelve adult female tree shrews were randomly divided into the Sham group and the TBI group, the Marmarou weight-drop impact method was used to establish a TBI model. Open field experiments were conducted after modeling to evaluate the motor ability of tree shrews on the 1st and 7th day. Moreover, magnetic resonance imaging (MRI), HE staining were performed to detect the imaging and morphology changes caused by trauma. The expression levels of inflammatory factors were detected by ELISA 7th days after TBI. Finally, immunofluorescence and Western Blot were used to detect the polarization characteristics of microglia and astrocytes. Open field experiments showed that the total motor distance of tree shrews in TBI group on the 1st and 7th days were significantly reduced. MRI showed high-density shadow in the parietal brain parenchyma, surrounded by low-density edema bands. HE staining showed in the TBI group, some neurons in the hippocampal region showed morphological abnormalities and nuclear condensation, glial cell proliferation, the arrangement of neurons in the cortical area is disordered, and the hierarchical structure is damaged. ELISA assays showed that the levels of pro-inflammatory factors in TBI group were significantly increased, while the anti-inflammatory factors’ expressions decreased. Finally, Immunofluorescence staining showed that, compared with Sham group, fluorescence signals of M1 phenotype macroglia marker CD86 and M2 phenotype marker CD206 in different regions of hippocampus in TBI group were significantly enhanced at 7 days after TBI. Western Blotting showed that compared with sham group, the expression of A1 phenotype marker Serping1and A2 phenotype marker Ptx3 protein of astrocytes in TBI group was significantly up-regulated. ELISA results showed that the expression levels of C3 and S100a10 in brain tissue of tree shrews in TBI group were significantly higher than those in Sham group. This study successfully constructed a TBI tree shrew model and confirmed that the polarization imbalance of microglia and astrocytes may form a neurotoxic microenvironment, ultimately causing brain tissue damage.

Objective This study aims to investigate the role and regulatory mechanisms of PTP1B in acute pancreatitis. Methods An in vitro AP cell model was established using Caerulin (CAE). Cells were subsequently treated with MSI-1436 (PTP1B inhibitor), ruscogenin (TXNIP inhibitor), N-acetyl-l-cysteine (NAC, ROS scavenger), and rotenone (mitochondrial complex I inhibitor), the levels of IL-1β and IL-18 in the cell supernatant were quantified using enzyme-linked immunosorbent assay (ELISA). Propidium iodide (PI) staining was visualized using fluorescence microscopy, and the fluorescence intensity of GSDMD was assessed via immunofluorescence. Western blotting was performed to detect the expression and activation of pro-caspase-1, cleaved-caspase-1, cleaved-caspase-4/5/11, AP-1, ASC, and NLRP3. Mitochondrial membrane potential was assessed using the JC-1 assay, complex I activity was determined with the MitoCheck™ Complex I Activity Assay Kit, and ROS levels were measured using a ROS detection kit. Results CAE treatment significantly increased the secretion of inflammatory cytokines IL-1β and IL-18 in pancreatic acinar cells. Expression levels of PTP1B, pyroptosis-related factors—including AP-1, ASC, cleaved-Caspase4, cleaved-Caspase5, cleaved-Caspase11, cleaved-Caspase1, and NLRP3—were significantly upregulated. However, inhibition of PTP1B and TXNIP markedly reduced inflammation and pyroptosis, suggesting that the PTP1B/TXNIP axis mediates CAE-induced cellular injury. Furthermore, suppression of ROS generation and mitochondrial complex I activity also attenuated CAE-induced inflammation and pyroptosis in pancreatic acinar cells. Conclusion Targeting PTP1B mitigates CAE-induced pyroptosis in pancreatic acinar cells by restoring NADH dehydrogenase activity and suppressing ROS/TXNIP/NLRP3 inflammasome activation.