This study aims to investigate the therapeutic effects of the Coptidis Rhizoma-Scutellariae Radix on cytokine storm secondary lung injury(CSSLI) induced by CpG1826 in mice, and to elucidate the potential molecular mechanisms by which its major active components, i.e., coptisine and wogonin, alleviate CSSLI by inhibiting the mitochondrial permeability transition pore(mPTP)/nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3) inflammasome pyroptosis pathway. In vivo, a mouse model of CSSLI was established by CpG1826 induction. Pulmonary edema was assessed by lung wet-to-dry weight ratio(W/D), lung injury was evaluated by hematoxylin-eosin(HE) staining, and ultrastructural changes in lung tissue were observed by transmission electron microscopy(TEM). The levels of interleukin(IL)-1β, high mobility group box 1 protein(HMGB1), IL-18, and IL-1α in bronchoalveolar lavage fluid were measured by enzyme-linked immunosorbent assay(ELISA). The results showed that the decoction of the Coptidis Rhizoma-Scutellariae Radix significantly reduced pulmonary edema, alleviated lung injury, and decreased the concentrations of related cytokines in BALF more effectively than either single herb alone, thereby improving CSSLI. In vitro, a CpG1826-induced CSSLI model was established in mouse alveolar macrophage MH-S cells. Calcein-AM quenching was used to screen for the most effective monomer components from the herb pair in inhibiting mPTP opening. Coptisine(5, 10, 20 μmol·L~(-1)) and wogonin(10, 20, 40 μmol·L~(-1)) markedly inhibited mPTP opening, with optimal effects and a clear dose-dependent pattern. These components suppressed mPTP opening, thereby reducing the release of mitochondrial DNA(mtDNA) and the accumulation of reactive oxygen species(ROS), effectively reversing the CpG1826-induced decrease in mitochondrial membrane potential(MMP). Further studies revealed that both coptisine and wogonin inhibited pyroptosis and downregulated the expression of key proteins in the NLRP3/Caspase-1/gasdermin D(GSDMD) pathway. In conclusion, the Coptidis Rhizoma-Scutellariae Radix improves CpG1826-induced CSSLI in mice, and this effect is associated with the inhibition of the mPTP/NLRP3 pyroptosis pathway, providing scientific evidence for its clinical application and further development.
Animals ; Mice ; Drugs, Chinese Herbal/administration & dosage* ; Pyroptosis/drug effects* ; NLR Family, Pyrin Domain-Containing 3 Protein/immunology* ; Male ; Lung Injury/immunology* ; Cytokines/immunology* ; Scutellaria baicalensis/chemistry* ; Oligodeoxyribonucleotides/adverse effects* ; Mice, Inbred C57BL ; Coptis chinensis

PURPOSE: To investigate the protective effect of sub-hypothermic mechanical perfusion combined with membrane lung oxygenation on ischemic hypoxic injury of yorkshire brain tissue caused by traumatic blood loss. METHODS: This article performed a random controlled trial. Brain tissue of 7 yorkshire was selected and divided into the sub-low temperature anterograde machine perfusion group (n = 4) and the blank control group (n = 3) using the random number table method. A yorkshire model of brain tissue injury induced by traumatic blood loss was established. Firstly, the perfusion temperature and blood oxygen saturation were monitored in real-time during the perfusion process. The number of red blood cells, hemoglobin content, NA⁺, K⁺, and Ca²⁺ ions concentrations and pH of the perfusate were detected. Following perfusion, we specifically examined the parietal lobe to assess its water content. The prefrontal cortex and hippocampus were then dissected for histological evaluation, allowing us to investigate potential regional differences in tissue injury. The blank control group was sampled directly before perfusion. All statistical analyses and graphs were performed using GraphPad Prism 8.0 Student t-test. All tests were two-sided, and p value of less than 0.05 was considered to indicate statistical significance. RESULTS: The contents of red blood cells and hemoglobin during perfusion were maintained at normal levels but more red blood cells were destroyed 3 h after the perfusion. The blood oxygen saturation of the perfusion group was maintained at 95% - 98%. NA⁺ and K⁺ concentrations were normal most of the time during perfusion but increased significantly at about 4 h. The Ca²⁺ concentration remained within the normal range at each period. Glucose levels were slightly higher than the baseline level. The pH of the perfusion solution was slightly lower at the beginning of perfusion, and then gradually increased to the normal level. The water content of brain tissue in the sub-low and docile perfusion group was 78.95% ± 0.39%, which was significantly higher than that in the control group (75.27% ± 0.55%, t = 10.49, p < 0.001), and the difference was statistically significant. Compared with the blank control group, the structure and morphology of pyramidal neurons in the prefrontal cortex and CA1 region of the hippocampal gyrus were similar, and their integrity was better. The structural integrity of granulosa neurons was destroyed and cell edema increased in the perfusion group compared with the blank control group. Immunofluorescence staining for glail fibrillary acidic protein and Iba1, markers of glial cells, revealed well-preserved cell structures in the perfusion group. While there were indications of abnormal cellular activity, the analysis showed no significant difference in axon thickness or integrity compared to the 1-h blank control group. CONCLUSIONS Mild hypothermic machine perfusion can improve ischemia and hypoxia injury of yorkshire brain tissue caused by traumatic blood loss and delay the necrosis and apoptosis of yorkshire brain tissue by continuous oxygen supply, maintaining ion homeostasis and reducing tissue metabolism level.
Animals ; Perfusion/methods* ; Disease Models, Animal ; Brain Injuries/etiology* ; Swine ; Male ; Hypothermia, Induced/methods*

OBJECTIVE: To analyze the Epstein-Barr virus (EBV) load in different lymphocyte subsets, as well as clinical characteristics and outcomes in patients with hematologic malignancies experiencing EBV reactivation. METHODS: Peripheral blood samples from patients were collected. B, T, and NK cells were isolated sorting with magnetic beads by flow cytometry. The EBV load in each subset was quantitated by real-time quantitative polymerase chain reaction (RT-qPCR). Clinical data were colleted from electronic medical records. Survival status was followed up through outpatient visits and telephone calls. Statistical analyses were performed using SPSS 25.0. RESULTS: A total of 39 patients with hematologic malignancies were included, among whom 35 patients had undergone allogeneic hematopoietic stem cell transplantation (allo-HSCT). The median time to EBV reactivation was 4.8 months (range: 1.7-57.1 months) after allo-HSCT. EBV was detected in B, T, and NK cells in 20 patients, in B and T cells in 11 patients, and only in B cells in 4 patients. In the 35 patients, the median EBV load in B cells was 2.19×10⁴ copies/ml, significantly higher than that in T cells (4.00×10³ copies/ml, P <0.01) and NK cells (2.85×10² copies/ml, P <0.01). Rituximab (RTX) was administered for 32 patients, resulting in EBV negativity in 32 patients with a median time of 8 days (range: 2-39 days). Post-treatment analysis of 13 patients showed EBV were all negative in B, T, and NK cells. In the four non-transplant patients, the median time to EBV reactivation was 35 days (range: 1-328 days) after diagnosis of the primary disease. EBV was detected in one or two subsets of B, T, or NK cells, but not simultaneously in all three subsets. These patients received a combination chemotherapy targeting at the primary disease, with 3 patients achieving EBV negativity, and the median time to be negative was 40 days (range: 13-75 days). CONCLUSION In hematologic malignancy patients after allo-HSCT, EBV reactivation commonly involves B, T, and NK cells, with a significantly higher viral load in B cells compared to T and NK cells. Rituximab is effective for EBV clearance. In non-transplant patients, EBV reactivation is restricted to one or two lymphocyte subsets, and clearance is slower, highlighting the need for prompt anti-tumor therapy.
Humans ; Hematologic Neoplasms/virology* ; Herpesvirus 4, Human/physiology* ; Epstein-Barr Virus Infections ; Hematopoietic Stem Cell Transplantation ; Virus Activation ; Lymphocyte Subsets/virology* ; Flow Cytometry ; Killer Cells, Natural/virology* ; Male ; Female ; B-Lymphocytes/virology* ; Viral Load ; Adult ; T-Lymphocytes/virology* ; Middle Aged

Objective To explore the abnormal expression of the MT-ND family in pan-cancers and its prognostic value.Methods Transcriptome expression and clinical data of 33 cancers were downloaded from the TCGA database,thereby analyzing the expression differences of the MT-ND family in tissuesof different types of cancers and normal tissues.The prognostic role of the MT-ND family in pan-cancers was explored by univariate Cox regression analysis and Kaplan-Meier survival analysis.Results The expression of the MT-ND family was upregulated in the kidney chromophobe,acute myeloid leukemia and thymoma,and downregulated in the remaining tumors.The expression of the MT-ND family was associated with the overall survival rate of different types of cancers.In addition,the MT-ND family were significantly correlated with the immune invasion subtypes,and were correlated with stromal cell invasion and tumor cell stemness to varying degrees.The expression of MT-ND4 was downregulated in brain lower grade glioma,which was a protective factor for prognosis;while the expression of MT-ND4 in thymoma was upregulated,which was a risk factor for prognosis.Conclusion Pan-cancer analysis has confirmed that the MT-ND family can predict the tumor prognosis,which provides new ideas and strategies for the precision treatment and prognostic management of cancer.

Graves'ophthalmopathy(GO)is a specific autoimmune disease occurring in orbital tissues and is closely related to hyperthyroidism caused by Graves'disease(GD).Its disabling and disfiguring features significantly impact the quality of life of patients.The exact mechanism of GO still remains to be fully elucidated.In recent years,gene sequencing and medical microbiology studies have shown that changes in the gut microbiota may play a role in the development and progression of GO,with gut dysbiosis altering immune system regulatory signals and causing immune damage to organs.Clarifying the correlation between gut microbiota and GO helps to understand the disease's pathogenic mechanism and provides a theoretical basis for the diagnosis and treatment of GO.This review summarizes the impact of gut microbiota dysbiosis in the pathogenesis of GO and promising therapeutic approaches,including research progress in aspects such as gut microbiota experiments,case studies,pathogenesis,and treatment strategies.

Objective To screen genes associated with poor prognosis of colorectal cancer(CRC)through bioinformatics analysis.Methods The gene expression profiles of GSE74602,GSE110223,GSE113513 and GSE141174 were obtained from Gene Expression Omnibus(GEO)database,including samples from 65 CRC tissues and 65 normal tissues.Differentially expressed genes(DEGs)between CRC tissues and normal tissues were screened out by GEO2R tool and Venn software,and the consistent genes were extracted from DEGs by Venn software.A protein-protein interaction(PPI)network was constructed by the STRING database to identify key genes,which were analyzed by Kaplan-Meier Plotter and GEPIA.Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis was conducted on the 13 selected genes.Results There were 171 DEGs obtained from the four datasets,including 148 up-regulated genes and 23 down-regulated genes.Up-regulated DEGs were enriched in the redox processes,bicarbonate transport,digestion,ion trans-membrane transport,and one-carbon metabolism;and down-regulated DEGs were enriched in the positive regulation of cell proliferation.The survival curve analysis showed that 30 of the 87 genes were significantly associated with poor survival prognosis.GEPIA showed that 13 of the 30 genes were highly expressed in CRC tissues compared to normal tissues,among which MYC and FGFR3 markedly enriched in the CRC pathway.Conclusion This study identifies that MYC and FGFR3 are significantly up-regulated in CRC and closely associated with poor prognosis,suggesting that they may serve as potential prognostic markers and therapeutic targets for CRC patients.

Objective To explore the abnormal expression of the MT-ND family in pan-cancers and its prognostic value.Methods Transcriptome expression and clinical data of 33 cancers were downloaded from the TCGA database,thereby analyzing the expression differences of the MT-ND family in tissuesof different types of cancers and normal tissues.The prognostic role of the MT-ND family in pan-cancers was explored by univariate Cox regression analysis and Kaplan-Meier survival analysis.Results The expression of the MT-ND family was upregulated in the kidney chromophobe,acute myeloid leukemia and thymoma,and downregulated in the remaining tumors.The expression of the MT-ND family was associated with the overall survival rate of different types of cancers.In addition,the MT-ND family were significantly correlated with the immune invasion subtypes,and were correlated with stromal cell invasion and tumor cell stemness to varying degrees.The expression of MT-ND4 was downregulated in brain lower grade glioma,which was a protective factor for prognosis;while the expression of MT-ND4 in thymoma was upregulated,which was a risk factor for prognosis.Conclusion Pan-cancer analysis has confirmed that the MT-ND family can predict the tumor prognosis,which provides new ideas and strategies for the precision treatment and prognostic management of cancer.

Objective To screen genes associated with poor prognosis of colorectal cancer(CRC)through bioinformatics analysis.Methods The gene expression profiles of GSE74602,GSE110223,GSE113513 and GSE141174 were obtained from Gene Expression Omnibus(GEO)database,including samples from 65 CRC tissues and 65 normal tissues.Differentially expressed genes(DEGs)between CRC tissues and normal tissues were screened out by GEO2R tool and Venn software,and the consistent genes were extracted from DEGs by Venn software.A protein-protein interaction(PPI)network was constructed by the STRING database to identify key genes,which were analyzed by Kaplan-Meier Plotter and GEPIA.Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis was conducted on the 13 selected genes.Results There were 171 DEGs obtained from the four datasets,including 148 up-regulated genes and 23 down-regulated genes.Up-regulated DEGs were enriched in the redox processes,bicarbonate transport,digestion,ion trans-membrane transport,and one-carbon metabolism;and down-regulated DEGs were enriched in the positive regulation of cell proliferation.The survival curve analysis showed that 30 of the 87 genes were significantly associated with poor survival prognosis.GEPIA showed that 13 of the 30 genes were highly expressed in CRC tissues compared to normal tissues,among which MYC and FGFR3 markedly enriched in the CRC pathway.Conclusion This study identifies that MYC and FGFR3 are significantly up-regulated in CRC and closely associated with poor prognosis,suggesting that they may serve as potential prognostic markers and therapeutic targets for CRC patients.

Ketamine,an antagonist of the glutamate N-methyl-D-aspartate(NMDA)receptor,is cur-rently one of the most widely abused psychoactive substances.Prolonged abuse can result in damages to various systems in the body,making it crucial to investigate the regulatory mechanism of ketamine addic-tion and screening related biomarkers.In this study,zebrafish embryos/larvae were initially exposed a-cutely to ketamine.Then,a ketamine addiction model was established in 6-month-old zebrafish through conditioned place preference(CPP)experiments.The zebrafish brain transcriptome was analyzed using RNA-seq,while qPCR and Western blotting were employed to detect the expression of key genes.Results revealed significant reductions in the spontaneous tail coiling,embryo hatching rate,and survival rate of zebrafish embryos in the ketamine-treated group compared to the control group.The distance moved also decreased significantly,from 1904.2 mm in the control group to 319.0 mm in the high dose of ketamine group(300 μmol/L).Conditional positional preference experiments demonstrated that the control ze-brafish did not exhibit significant changes in activity in the CPP tank.In contrast,the ketamine-treated group increased their activity time in the light zone of the tank from 385.2 s before training to 706.4 s af-ter training,representing a 26.8％increase(***P＜0.001).This suggests a preference for ketamine stimulation in zebrafish.KEGG analysis indicated enrichment of differentially expressed genes in the neu-roactive ligand-receptor interaction pathway in the ketamine-treated samples.GSEA analysis further re-veals a significant upregulation of the glutamatergic synapse pathway(NES=1.5).In addition,compared with the control group,the mRNA levels of Grin2b and Gria2 in the ketamine group increased by 4.6 and 1.4 times,respectively,while the protein levels increased by 2.0 and 1.4 times,respectively.These findings suggest that ketamine can induce addiction in zebrafish,potentially through upregulation of the glutamatergic synaptic pathway.

OBJECTIVE: Despite the combination of Scutellaria barbata D. Don and Scleromitrion diffusum (Willd.) R.J. Wang (SB-SD) being a recognized Chinese medicinal herbal pair that is commonly used in the treatment of ovarian cancer, there is a poor understanding of their pharmacological mechanisms. This study examines the antitumor properties and potential mechanisms of SB-SD on human ovarian cancer A2780 cells through a multi-omics approach, establishing a pharmacological basis for clinical utilization. METHODS: A range of mass ratios and reagents were used in the hot reflux extraction of SB-SD. The inhibitory effect of the SB-SD extracts on A2780 cell proliferation was assessed using the cell-counting kit 8 assay. A zebrafish tumor implantation model was used to evaluate the effects of SB-SD extracts on tumor growth and metastasis in vivo. Transcriptomics and proteomics were used to investigate alterations in biological pathways in A2780 cells after treatment with different concentrations of SB-SD extract. Cell cycle, cell apoptosis, intracellular free iron concentration, intracellular reactive oxygen species (ROS) concentration, malondialdehyde (MDA), and mitochondrial membrane potential were measured. Real-time quantitative reverse transcription polymerase chain reaction and Western blotting were utilized to investigate the effects of heme catabolism and ferritinophagy on ferroptosis induced by SB-SD extract in A2780 cells. RESULTS: The 70% ethanol extract of SB-SD (a mass ratio of 4:1) inhibited A2780 cell proliferation significantly with a half maximal inhibitory concentration of 660 μg/mL in a concentration- and time-dependent manner. Moreover, it effectively suppressed tumor growth and metastasis in a zebrafish tumor implantation model. SB-SD extract induced the accumulation of free iron, ROS, MDA, and mitochondrial damage in A2780 cells. The mechanisms might involve the upregulated expression of ferritinophagy-related genes microtubule-associated protein 1 light chain 3, autophagy-related gene 5, and nuclear receptor coactivator 4. CONCLUSION SB-SD extract effectively inhibited the development of ovarian cancer both in vitro and in vivo. Its mechanism of action involved inducing ferroptosis by facilitating heme catabolism and ferritinophagy. This herbal pair holds promise as a potential therapeutic option for ovarian cancer treatment and may be utilized in combination with routine treatment to improve the treatment outcomes of ovarian cancer patients. Please cite this article as: Wang Z, Liu M, Li GX, Zhang L, Ding KY, Li SQ, Gao BQ, Chen P, Choe HC, Xia LY, Yang YT, Liu Y, Sui X, Ma JN, Zhang L. A herbal pair of Scutellaria barbata D. Don and Scleromitrion diffusum (Willd.) R.J. Wang induced ferroptosis in ovarian cancer A2780 cells via inducing heme catabolism and ferritinophagy. J Integr Med. 2024; 22(6): 666-683.
Ferroptosis/drug effects* ; Female ; Humans ; Animals ; Scutellaria/chemistry* ; Ovarian Neoplasms/genetics* ; Zebrafish ; Cell Line, Tumor ; Ferritins/genetics* ; Plant Extracts/pharmacology* ; Heme/metabolism* ; Drugs, Chinese Herbal/pharmacology* ; Cell Proliferation/drug effects* ; Reactive Oxygen Species/metabolism* ; Antineoplastic Agents, Phytogenic/pharmacology* ; Autophagy/drug effects* ; Apoptosis/drug effects*