Objective To investigate the effects of cornuside on intestinal injury in rats with septic shock,and clarify its possible mechanism.Methods SD rats were randomly divided into normal control group,model group,low-,medium-,and high-dose comecarpine glycosides groups,and TREM1 inhibitor(LR12)group.HE staining was used to observe the pathological injury of small intestinal mucosa.The levels of D-lactic acid(D-LA)and diamine oxidase(DAO)in serum and secretory immunoglobulin(sIg A)in small intestine were detected by ELISA.Intestinal mucosal permeability was detected by fluorescein isothiocyanate-dextran(FITC-D)tracer method.ELISA was used to detect the levels of interferon(IFN)-γ,tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-10 and arginase(Arg)-1 in serum.The polarization of macrophages in small intestinal tissue was detected by flow cytometry.Western blot was used to detect the protein expression levels of triggering receptor expressed on myeloid cells 1(TREM1),CD86 and CD206 in small intestine.Results Compared with the normal control group,the model group had serious pathological injury of the small intestinal mucosa,and the serum levels of D-LA,DAO,FITC-D,IFN-γ,TNF-α,and IL-1β significantly increased(P<0.05),while the levels of sIg A,IL-10,and Arg-1 significantly decreased(P<0.05).The M1/M2 ratio of macrophages and the expression levels of TREM1 and CD86 proteins in the small intestine tissue significantly increased(P<0.05),while the expression level of CD206 protein significantly decreased(P<0.05).Compared with the model group,the small intestinal mucosal injury of the rats in each dose cornuside group and LR12 group significantly improved,and the serum levels of D-LA,DAO,FITC-D,IFN-γ,TNF-α,and IL-1β significantly decreased(P<0.05),while the levels of sIg A,IL-10,and Arg-1 significantly increased(P<0.05).The M1/M2 ratio of macrophages and the expression levels of TREM1 and CD86 proteins in the small intestine tissue significantly decreased(P<0.05),while the expression level of CD206 protein significantly increased(P<0.05).Conclusion Cornuside can reduce intestinal injury in rats with septic shock,and the mechanism may be related to inhibiting TREM1-mediated M1 polarization of macrophages.

Objective To investigate the effects of cornuside on intestinal injury in rats with septic shock,and clarify its possible mechanism.Methods SD rats were randomly divided into normal control group,model group,low-,medium-,and high-dose comecarpine glycosides groups,and TREM1 inhibitor(LR12)group.HE staining was used to observe the pathological injury of small intestinal mucosa.The levels of D-lactic acid(D-LA)and diamine oxidase(DAO)in serum and secretory immunoglobulin(sIg A)in small intestine were detected by ELISA.Intestinal mucosal permeability was detected by fluorescein isothiocyanate-dextran(FITC-D)tracer method.ELISA was used to detect the levels of interferon(IFN)-γ,tumor necrosis factor(TNF)-α,interleukin(IL)-1β,IL-10 and arginase(Arg)-1 in serum.The polarization of macrophages in small intestinal tissue was detected by flow cytometry.Western blot was used to detect the protein expression levels of triggering receptor expressed on myeloid cells 1(TREM1),CD86 and CD206 in small intestine.Results Compared with the normal control group,the model group had serious pathological injury of the small intestinal mucosa,and the serum levels of D-LA,DAO,FITC-D,IFN-γ,TNF-α,and IL-1β significantly increased(P<0.05),while the levels of sIg A,IL-10,and Arg-1 significantly decreased(P<0.05).The M1/M2 ratio of macrophages and the expression levels of TREM1 and CD86 proteins in the small intestine tissue significantly increased(P<0.05),while the expression level of CD206 protein significantly decreased(P<0.05).Compared with the model group,the small intestinal mucosal injury of the rats in each dose cornuside group and LR12 group significantly improved,and the serum levels of D-LA,DAO,FITC-D,IFN-γ,TNF-α,and IL-1β significantly decreased(P<0.05),while the levels of sIg A,IL-10,and Arg-1 significantly increased(P<0.05).The M1/M2 ratio of macrophages and the expression levels of TREM1 and CD86 proteins in the small intestine tissue significantly decreased(P<0.05),while the expression level of CD206 protein significantly increased(P<0.05).Conclusion Cornuside can reduce intestinal injury in rats with septic shock,and the mechanism may be related to inhibiting TREM1-mediated M1 polarization of macrophages.

Objective:This study aimed to evaluate the effect of early tocilizumab intervention to relieve cytokine release syndrome (CRS) following chimeric antigen receptor T cell (CAR-T) therapy.Methods:Twenty-two patients with acute lymphoblastic leukemia who received tocilizumab to relieve CRS response after CAR-T cell infusion in our research center from October 2015 to July 2021 were retrospectively analyzed. According to the timing of tocilizumab intervention, patients were divided into the conventional and early intervention groups. Patients who received tocilizumab treatment after sustained high fever for 4 h were included in the early intervention group. The clinical data, CRS grade, and event-free survival (EFS) between the two groups were evaluated.Results:Compared with patients who used tocilizumab after severe CRS, no patients in the early intervention group died from CRS, and there was no increased risk of neurotoxicity. Eleven patients (84.62%) achieved complete remission with minimal residual lesions. The median EFS of patients in the early intervention and conventional groups was 2 (95% CI 0-5) and 7 (95% CI 3-11) months, respectively. Conclusion:Early tocilizumab intervention in patients with CRS reduces severe CRS and provides a more optimized therapeutic strategy for CRS caused by CAR-T cell therapy.

Objective:To analyze the diagnostic value of metabolic makers in cerebrospinal fluid in advanced lung adenocarcinoma patients with leptomeningeal metastases (LM) .Methods:A total of 46 cerebrospinal fluid samples (LM group) from lung adenocarcinoma patients with LM admitted to Nanjing Drum Tower Hospital Affiliated to Nanjing University Medical School from December 2019 to December 2021 were collected, and 48 cerebrospinal fluid samples (control group) from patients with benign neurological diseases during the same period were collected. Metabolomics analysis of cerebrospinal fluid was carried out by high-performance liquid chromatography-mass spectrometry. Principle component analysis (PCA) and orthogonal to partial least squares discriminant analysis (OPLS-DA) were used for modeling. Multi-criteria assessment was used to identify the different metabolites between the two groups. Receiver operating characteristic (ROC) curve, pathway enrichment analysis and other methods were used to screen metabolites and pathways related to LM from lung adenocarcinoma.Results:There were no statistically significant differences in the proportions of age ( Z=-0.41, P=0.210) , gender ( χ2=1.19, P=0.275) , history of smoking ( χ2=2.86, P=0.091) , Karnofsky performance status score ( χ2=0.65, P=0.419) and increased intracranial pressure ( χ2=0.65, P=0.419) between the LM group and control group. The models of PCA (R2X was 0.608 and 0.583, Q2 was 0.462 and 0.513 in electrospray ion positive and negative modes, respectively) and OPLS-DA (R2Y was 0.967 and 0.889, Q2 was 0.959 and 0.852 in electrospray ion positive and negative modes, respectively) showed that the overall data quality was good. Meanwhile, the model interpretation rate and prediction rate were effective. The permutation tests duplicated for 200 times and showed no over-fitting of the established model. The metabolic profiles of the two groups were significantly different. A total of 30 endogenously differential metabolites were screened by using multi-criteria assessment. Six potential biomarkers with larger area under the curve (AUC) were identified through ROC curve analysis, including tyrosine (AUC=0.967, 95% CI: 0.906-1.000) , phenylalanine (AUC=0.992, 95% CI: 0.973-1.000) , pyruvate (AUC=0.976, 95% CI: 0.935-1.000) , tryptophan (AUC=0.935, 95% CI: 0.880-0.973) , glucose (AUC=0.932, 95% CI: 0.880-0.975) and adenosine monophosphate (AUC=0.993, 95% CI: 0.987-1.000) . The 30 selected differential metabolites were enriched and analyzed for metabolic pathways, and 20 relevant metabolic pathways were matched. Among them, the four metabolic pathways most likely to cause changes in metabolites were glycolysis and glucose metabolic synthesis, pyruvate metabolism, phenylalanine metabolism, phenylalanine, tyrosine and tryptophan biosynthesis. Conclusion:Untargeted metabolomics analysis can effectively screen specific cerebrospinal fluid metabolites in lung adenocarcinoma patients with LM. Six potential metabolites such as tyrosine, phenylalanine, pyruvate, tryptophan, adenosine monophosphate, glucose and their metabolic pathways may be involved in the pathogenesis of LM from lung adenocarcinoma.

Objective Drug resistance is the main cause of chemotherapy failure in hepatocellular carcinoma (HCC), and thioredoxin reductase 1 (TXNRD1), as a major influencing factor for reactive oxygen species (ROS) metabolism, has been proven to be associated with the poor prognosis of patients with HCC. This study aims to explore the role of TXNRD1 in the mechanism of multidrug resistance in HCC. Methods BEL/FU cells in BEL-7402 cell line were selected as the multidrug-resistant cell line. The siRNA was used for the intervention of TXNRD1 expression; quantitative real-time PCR and Western blotting were used to measure the expression of TXNRD1; CCK-8 assay and flow cytometry were used to evaluate the effect of TXNRD1 on hepatocyte ROS accumulation, resistance to 5-fluorouracil (5-Fu) and doxorubicin (DOX), and apoptosis in vitro; a xenograft tumor model was established to investigate the effect of auranofin (AUR) on drug resistance in vivo. The two-independent-samples t test was used for comparison of continuous data between two groups. Results As a multidrug-resistant HCC cell line, BEL/Fu showed high mRNA and protein expression levels of TXNRD1 (both P < 0.05). Compared with 5-Fu or DOX treatment alone, the TXNRD1 inhibitor AUR combined with 5-Fu or DOX had had a significant reduction in the number of colony formation ( P < 0.01) and a significant increase in apoptosis ratio ( P < 0.001). The ROS scavenger N-acetylcysteine (NAC) significantly weakened the effect of TXNRD1 knockdown by siRNA on the drug resistance of BEL/Fu cells, and the application of NAC effectively reduced the apoptosis ratio of cells after siRNA interference ( P < 0.001). Animal experiments also confirmed that compared with the nude mice treated with 5-Fu alone, the nude mice treated with 5-Fu and AUR had a significantly lower tumor mass ( P < 0.001) and a significantly smaller tumor volume ( P < 0.001). Conclusion TXNRD1 plays an important role in the drug resistance of HCC, and inhibition of its level in cells can effectively improve drug resistance. As a TXNRD1 inhibitor, AUR has great application prospects in the multimodality therapy for HCC.

The eukaryotic genome is folded into higher-order conformation accompanied with constrained dynamics for coordinated genome functions. However, the molecular machinery underlying these hierarchically organized three-dimensional (3D) chromatin architecture and dynamics remains poorly understood. Here by combining imaging and sequencing, we studied the role of lamin B1 in chromatin architecture and dynamics. We found that lamin B1 depletion leads to detachment of lamina-associated domains (LADs) from the nuclear periphery accompanied with global chromatin redistribution and decompaction. Consequently, the inter-chromosomal as well as inter-compartment interactions are increased, but the structure of topologically associating domains (TADs) is not affected. Using live-cell genomic loci tracking, we further proved that depletion of lamin B1 leads to increased chromatin dynamics, owing to chromatin decompaction and redistribution toward nucleoplasm. Taken together, our data suggest that lamin B1 and chromatin interactions at the nuclear periphery promote LAD maintenance, chromatin compaction, genomic compartmentalization into chromosome territories and A/B compartments and confine chromatin dynamics, supporting their crucial roles in chromatin higher-order structure and chromatin dynamics.
Chromatin ; Chromosomes ; Genome ; Humans ; Lamin Type B/genetics*

BACKGROUND: Tumor markers (TM) in cerebrospinal fluid (CSF) are useful for diagnosing leptomeningeal metastasis (LM). It has not been fully exploited the diagnostic possibilities of the CSF levels since the basic fact that the TM concentration of CSF depends strongly upon the serum levels as well as upon the condition of the blood brain barrier (BBB). To analyze the intrathecal TM synthesis and evaluate the integrity of BBB can be helpful for the definitive diagnosis of LM. Therefore, the aim of this study was to further explore the clinical value of intrathecal TM synthesis and BBB in the diagnosis for the lung cancer patients with LM. METHODS: Twenty-five lung cancer patients with LM and 57 patients with nonmalignant neurological diseases (NMNDs) admitted to Nanjing Drum Tower Hospital from December 2016 to March 2020 were included. We compared the integrity of BBB and intrathecal TM synthesis between two groups, analyzed the correlation of CSF TM between the detection and intrathecal synthesis, and evaluated serial CSF cytology, the integrity of BBB and intrathecal TM synthesis when intrathecal chemotherapy for one patient. RESULTS: Ninety-four percent LM patients showed the dysfunction of BBB, and all LM patients showed at least one intrathecal synthesized TM in CSF. In one patient, the CSF cytology was negative for the first time, but LM was eventually diagnosed based on the the intrathecal TM synthesis and positive CSF cytology of repeated lumbar puncture. In LM group, no correlation was observed between the detection and intrathecal synthesized TM in CSF. In the control group, only 3.5% (2/57) NMNDs patients had the dysfunction of BBB and no patients had intrathecal TM synthesis, both the differences of which were statistically significant (P<0.05). Finally, evaluating the CSF cytology, integrity of BBB and intrathecal TM synthesis can be used to assess the intracranial treatment effect. Moreover, intrathecal TM synthesis changes earlier than cytology. CONCLUSIONS The evaluation of intrathecal TM synthesis and integrity of BBB are novel clinical diagnostic tools. In addition, serial measurement of intrathecal synthesized TM may play an important role in monitoring efficacy of lung cancer patients with LM, which is worthy of further promotion and clinical application.