Matrine protects against experimental autoimmune encephalomyelitis through modulating microglial ferroptosis.

Multiple sclerosis (MS) is an inflammatory demyelination neurodegenerative disease of the central nervous system (CNS). Ferroptosis has been implicated in a range of brain disorders, and iron-loaded microglia are frequently found in affected brain regions. However, the molecular mechanisms linking ferroptosis with MS have not been well-defined. The present study seeks to bridge this gap and investigate the impact of matrine (MAT), a herbal medicine with immunomodulatory capacities, on the regulation of oxidative stress and ferroptosis in the CNS of mice with experimental autoimmune encephalomyelitis (EAE), an animal model of MS. CNS of EAE mice contained elevated levels of ferroptosis-related molecules, e.g., MDA, LPCAT3 and PTGS2, but decreased expression of antioxidant molecules, including GSH and SOD, GPX4 and SLC7A11. This pathogenic process was reversed by MAT treatment, together with significant reduction of disease severity and CNS inflammatory demyelination. Furthermore, the expression of PTGS2 and LOX was largely increased in microglia of EAE mice, accompanied with increased production of IL-6 and TNF-α, indicating a proinflammatory phenotype of microglia that undergo oxidative stress/ferroptosis, and their expression was significantly reduced after MAT treatment. Together, our results indicate that ferroptosis/inflammation plays an important role in the pathogenesis of CNS autoimmunity, and inhibiting ferroptosis-induced microglial activation/inflammation could be a novel mechanism underlying the therapeutic effects of MAT on CNS inflammatory demyelination in EAE.

Sewage sludge pyrolysis 'kills two birds with one stone': Biochar synergies with persulfate for pollutants removal and energy recovery.

The disposal and resource utilization of sewage sludge (SS) have always been significant challenges for environmental protection. This study employed straightforward pyrolysis to prepare iron-containing sludge biochar (SBC) used as a catalyst and to recover bio-oil used as fuel energy. The results indicated that SBC-700 could effectively activate persulfate (PS) to remove 97.2% of 2,4-dichlorophenol (2,4-DCP) within 60 min. Benefiting from the appropriate iron content, oxygen-containing functional groups and defective structures provide abundant active sites. Meanwhile, SBC-700 exhibits good stability and reusability in cyclic tests and can be easily recovered by magnetic separation. The role of non-radicals is emphasized in the SBC-700/PS system, and in particular, single linear oxygen (1O2) is proposed to be the dominant reactive oxygen. The bio-oil, a byproduct of pyrolysis, exhibits a higher heating value (HHV) of about 30 MJ/kg, with H/C and O/C ratios comparable to those of biodiesel. The energy recovery rate of the SS pyrolysis system was calculated at 80.5% with a lower input cost. In conclusion, this investigation offers a low-energy consumption and sustainable strategy for the resource utilization of SS while simultaneously degrading contaminants.

Qualitative and quantitative analysis of electrons donated by pollutants in electron transfer-based oxidation system: Electrochemical measurement and theoretical calculations.

In order to gain a profound understanding of the fate of pollutants in advanced oxidation processes (AOPs), this study analyzed the electron contribution of pollutants qualitatively and quantitatively which rarely reported before. The rich electron transfer system was constructed by mesoporous carbon nitride (MCN) coupling with persulfate (PS) driven by visible light and the sulfanilamide antibiotics (SULs) were used as target contaminants. Firstly, the qualitative analysis of electron transfer in the system was confirmed systematically. The electron flow direction tested by i-t curves indicated that PS absorbed electrons, while SULs released electrons. The flow rate of electrons was also accelerated after the addition of SULs. The fitting curve between the kinetics and the peak potential difference tested by CV curve showed that the larger potential difference, the slower rate of oxidative degradation. Secondly, the quantification of electron transfer was achieved through theoretical calculations to simulate the interactions of the 'catalyst-oxidant-antibiotic' system. After the addition of SULs, the adsorption energy of the 'catalyst-oxidant-antibiotic' system was enhanced and the bond length of the peroxide bond was stretched. Notably, the electron transfer analysis results showed that the charge of SULs was around 0.032-0.056e, indicating that SULs pollutants played the role of electron contributors in the system. The oxidative degradation pathway included the direct cracking of S-N bond, shedding of marginal groups, ring-opening and hydroxyl addition reaction. This study clarified the electronic contribution of SULs in the oxidation system, providing necessary theoretical supplement for the analysis of the transformation of pollutants in AOPs.

Matrine Mediated Immune Protection in MS by Regulating Gut Microbiota and Production of SCFAs.

There is clearly an unmet need for more effective and safer treatments for multiple sclerosis (MS). Our previous studies showed a significant therapeutic effect of matrine, a monomer of traditional herbal medicine, on experimental autoimmune encephalomyelitis (EAE) mice. To explore the mechanism of matrine action, we used 16S rRNA sequencing technology to determine the gut microbes in matrine-treated EAE mice and controls. The concentrations of short-chain fatty acids (SCFAs) were then tested by metabonomics. Finally, we established pseudo-sterile mice and transplanted into them fecal microbiota, which had been obtained from the high-dose matrine-treated EAE mice to test the effects of matrine. The results showed that matrine could restore the diversity of gut microbiota and promote the production of SCFAs in EAE mice. Transplantation of fecal microbiota from matrine-treated mice significantly alleviated EAE severity, reduced CNS inflammatory infiltration and demyelination, and decreased the level of IL-17 but increased IL-10 in sera of mice. In conclusion, matrine treatment can regulate gut microbiota and metabolites and halt the progression of MS.

Reconstruction of massive bone defects after femoral tumor resection using two new-designed 3D-printed intercalary prostheses: a clinical analytic study with the cooperative utilization of multiple technologies.

BACKGROUND: To reconstruct massive bone defects of the femoral diaphysis and proximal end with limited bilateral cortical bone after joint-preserving musculoskeletal tumor resections, two novel 3D-printed customized intercalary femoral prostheses were applied. METHODS: A series of nine patients with malignancies who received these novel 3D-printed prostheses were retrospectively studied between July 2018 and November 2021. The proximal and diaphyseal femur was divided into three regions of interest (ROIs) according to anatomic landmarks, and anatomic measurements were conducted on 50 computed tomography images showing normal femurs. Based on the individual implant-involved ROIs, osteotomy level, and anatomical and biomechanical features, two alternative 3D-printed prostheses were designed. In each patient, Hounsfield Unit (HU) value thresholding and finite element analysis were conducted to identify the bone trabecula and calcar femorale and to determine the stress distribution, respectively. We described the characteristics of each prosthesis and surgical procedure and recorded the intraoperative data. All patients underwent regular postoperative follow-up, in which the clinical, functional and radiographical outcomes were evaluated. RESULTS: With the ROI division and radiographic measurements, insufficient bilateral cortical bones for anchoring the traditional stem were verified in the normal proximal femur. Therefore, two 3D-printed intercalary endoprostheses, a Type A prosthesis with a proximal curved stem and a Type B prosthesis with a proximal anchorage-slot and corresponding locking screws, were designed. Based on HU value thresholding and finite element analysis, the 3D-printed proximal stems in all prostheses maximally preserved the trabecular bone and calcar femorale and optimized the biomechanical distribution, as did the proximal screws. With the 3D-printed osteotomy guide plates and reaming guide plates, all patients underwent the operation uneventfully with a satisfactory duration (325.00 ± 62.60 min) and bleeding volume (922.22 ± 222.36 ml). In the follow-up, Harris Hip and Musculoskeletal Tumor Society scores were ameliorated after surgery (P < 0.001 and P < 0.001, respectively), reliable bone ingrowth was observed, and no major complications occurred. CONCLUSIONS: Two novel 3D-printed femoral intercalary prostheses, which achieved acceptable overall postoperative outcomes, were used as appropriate alternatives for oncologic patients with massive bone defects and limited residual bone and increased the opportunities for joint-preserving tumor resection. Several scientific methodologies utilized in this study may promote the clinical design proposals of 3D-printed implants.

Persulfate activation by sludge-derived biochar for efficient degradation of 2,4-dichlorophenol: performance and mechanism.

Porous sludge biochar (PSDBC) and zero-valent iron (ZVI) supported on porous sludge biochar composite (ZVI@PSDBC) were synthesized using municipal sludge through pyrolysis under N2 atmosphere, which manifested upgraded performance in persulfate (PS) activation for 2,4-dichlorophenol (2,4-DCP) degradation. The 2,4-DCP (50 mg/L) could be almost completely removed within 20 min under relatively low PS dosage (0.5 mmol/L) in both PSDBC/PS and ZVI@PSDBC/PS systems, and the mineralization rate could respectively approach 73.7% and 91.6% in 60 min. Combined with a scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS) characterization and electron spin-resonance (ESR) detection, electrochemical analysis, the radical and non-radical pathways in the catalytic systems were discussed. Graphitized structure and superior conductivity made PSDBC and ZVI@PSDBC not only act as electron donors in PS activation to create radicals (mainly SO4·- and ·OH), but also as "mediators" to facilitate the direct electron transfer from 2,4-DCP to the catalysts-PS complexes. The C=O groups of PSDBC and ZVI@PSDBC aided in the production of 1O2. Meanwhile, zero-valent iron nanoparticles promoted the formation of radicals as the reactive sites of PS, resulting in the most effective 2,4-DCP degradation in the ZVI@PSDBC/PS system. The stability and practicability of sludge biochar materials had been demonstrated in reusability and actual wastewater experiments. The findings provided a promising way for the reuse of municipal sludge and effective PS activation in wastewater treatment.

Ferroptosis as a mechanism of oligodendrocyte loss and demyelination in experimental autoimmune encephalomyelitis.

Ferroptosis, distinct from necrosis, autophagy and apoptosis, is a unique form of regulated cell death，and is a potential pathogenic mechanism of neuronal loss and defunction in many neurodegenerative disorders. Recent studies have shown a presence of iron deposition in the central nervous system (CNS) of patients with multiple sclerosis (MS). However, whether ferroptosis is involved in the pathogenesis of MS remains unclear. In the present study, we tested certain classical biomarkers of ferroptosis in the spinal cord of mice with experimental autoimmune encephalomyelitis (EAE), an animal model of MS, to substantiate the relationship between ferroptosis and oligodendrocyte (OL) loss and demyelination. Our results revealed decreased levels of key molecules in glutathione antioxidant mechanisms, including system xC (xCT) and glutathione peroxidase 4 (GPX4) in spinal cord of EAE mice, with evident lipid peroxidation in OLs. Moreover, transferrin receptor and ferritinophagy further catalyzed the generation of lipid reactive oxygen species through the fenton reaction, which induced OL death and demyelination at disease peak of EAE. This phenomenon was largely reversed by administering Fer-1, an inhibitor of ferritin phagocytosis, further validating the key role of ferritin phagocytosis in EAE. Taken together, these findings demonstrate that OL loss and demyelination may be induced in EAE through, at least in part, a mechanism of ferroptosis.

Effects of compound small peptides of Chinese medicine on intestinal immunity and cecal intestinal flora in CTX immunosuppressed mice.

The study was designed to explore the improvement effect of CSPCM (compound small peptide of Chinese medicine) on intestinal immunity and microflora through the treatment of different doses of CSPCM. A total of 100 male Kunming mice were weighed and divided into five groups, namely, group A (control group), group B (model group), group C (0.1 g/kg·bw CSPCM), group D (0.2 g/kg·bw CSPCM), and group E (0.4 g/kg·bw CSPCM). The use of CTX (cyclophosphamide) caused a series of negative effects: the secretion of IL-2, IL-22, TNF-α, sIgA, length of the villi, and the area of Pey's node were significantly reduced (P < 0.05); the depth of crypt and the percent of CD3+ and CD4+ cells were significantly increased (P < 0.05); the cecal flora taxa decreased; the abundance of Firmicutes and Lactobacillus increased; and the abundance of Bacteroidetes, Deferribacteres, Proteobacteria, Mucispirillum, Bacteroides, and Flexisprra decreased. The addition of CSPCM improved the secretion of cytokines and the development of intestinal villi, crypts, and Pey's node. The number of CD3+ and CD4+ cells in groups C, D, and E was significantly higher than that in group B (P < 0.05). Compared with group B, the abundance of Firmicutes in groups C, D, and E was decreased, and the Bacteroidetes, Deferribacteres, and Proteobacteria increased. The abundance of Lactobacillus decreased, while that of Mucispirillum, Bacteroides, and Flexisprra increased. It is concluded that cyclophosphamide is extremely destructive to the intestinal area and has a great negative impact on the development of the small intestine, the intestinal immune system, and the intestinal flora. The CSPCM can improve the negative effects of CTX.

Salvia miltiorrhiza polysaccharides ameliorates Staphylococcus aureus-induced mastitis in rats by inhibiting activation of the NF-κB and MAPK signaling pathways.

The lactation capacity of dairy cows is critical to the productivity of the animals. Mastitis is a disease that directly affects the lactation capacity of cows. Staphylococcus aureus (S. aureus) is one of the most important pathogens that causes mastitis in dairy cows. The anti-inflammatory effect of Salvia miltiorrhiza polysaccharides (SMPs) has been demonstrated in mice and chickens. However, the effectiveness of SMPs in preventing and treating mastitis is unclear. Therefore, the purpose of this study was to explore the protective effect and mechanism of SMPs on mastitis caused by S. aureus. S. aureus was used to induce mastitis in rats, and three doses of SMPs (87.5, 175, 350 mg/kg, BW/d) were administered as treatments. The bacterial load, histopathology, and myeloperoxidase (MPO) and N-acetyl-ß-D-glucosaminidase (NAGase) activities of mammary glands were observed and measured. Cytokines, including interleukin (IL)-1ß, interleukin (IL)-6, and tumor necrosis factor α (TNF-α), were examined by qRT-PCR and ELISA. Key proteins in the NF-κB and MAPK signaling pathways were analyzed by Western blotting. The results showed that SMP supplementation could significantly reduce the colonization of S. aureus and the recruitment of inflammatory cells in mammary glands. S. aureus-induced gene transcription and protein expression of IL-1ß, IL-6, and TNF-α were significantly suppressed in mammary glands. In addition, the increase in NF-κB and MAPK protein phosphorylation was inhibited by SMPs. These results revealed that supplementation with SMPs protected the mammary gland of rats against damage caused by S. aureus and alleviated the inflammatory response. This study provides a certain experimental basis for the treatment of S. aureus-induced mastitis with SMPs in the future.

Matrine inhibits the Wnt3a/ß-catenin/TCF7L2 signaling pathway in experimental autoimmune encephalomyelitis.

Oligodendrocyte (OL) death and remyelination failure lead to progressive neurological deficits in multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE). Matrine (MAT), a quinolizidine alkaloid component derived from the root of Sophora flavescens, has the capacity to effectively inhibit central nervous system (CNS) inflammation and to promote neuroregeneration. In the present study we explored its regulatory mechanism on the Wnt/ß-catenin/TCF7L2 pathway, a negative modulator for myelination, in MOG35--55 peptide-induced EAE. Our results clearly indicate that MAT treatment reduced the activation of Wnt3a and ß-catenin in the CNS of EAE mice, accompanied by the activation of GSK3ß and decreased expression of cyclin D1 and Axin2, two target genes of the Wnt3a/ß-catenin pathway. In addition, MAT increased OL maturation and myelination, as evidenced by the decreased number of NG2+Olig2+ cells and the increased numbers of MBP+ and CC1+Olig2+ cells. Taken together, these findings indicate that MAT treatment promoted the maturation of OLs and myelin repair, which is closely related to the modulation of the Wnt/ß-catenin/TCF7L2 signaling pathway.

Synergetic mechanism of defective g-C3N4 activated persulfate on removal of antibiotics and resistant bacteria: ROSs transformation, electron transfer and noncovalent interaction.

The environmental hazards of antibiotics and the resulting antibiotic-resistant bacteria (ARB) have attracted more and more attention. In this study, an efficient synergistic system constructed by vacancy g-C3N4 (CNV0.8) and persulfate (PS) showed excellent oxidation performance to degrade aztreonam (AZT) and Escherichia coli (E. coli) screened from wastewater treatment plant (WWTP), as the typical ß-lactam antibiotic and ARB. As the recombination of electron and hole was effectively inhibited and the interaction with PS was enhanced after the introduction of defects, CNV0.8 showed superior PS activation ability compared with bulk-g-C3N4 (BCN). The synergistic mechanism was systematically analyzed at three levels step by step. Firstly, the conversion of reactive oxygen radicals (ROSs) was studied using electron spin resonance (ESR) and quenching experiments. Then based on the DFT simulation, the enhancement of adsorption energy between catalysts and PS from -8.924 eV (BCN) to -11.190 eV (CNV0.8) and the elongation of O-O bond in PS (from 1.496 Å to 1.505 Å) indicated CNV0.8 had better activation performance for PS compared with BCN. The electron transfer results observed by deformation charge density showed that more electrons could be transferred from the CNV0.8 layer to the surrounding of PS for its own activation in the synergistic mechanism. Thirdly, the noncovalent interaction of PS/CNV0.8 belonged to the region of van der Waals force which was defined by the reduced density gradient (RDG) analysis. The intermediate products in the degradation of AZT were first studied in detail using Fukui function calculations and HPLC-QTOF-MS analysis. Subsequently, the environmental practicability of the oxidation system was investigated through wastewater simulation. This research provides a possible strategy for the effective removal of micropollutants and promotes the development of the sulfate radical-advanced oxidation processes (SR-AOPs) in the field of wastewater treatment.

125I seeds inhibit proliferation and promote apoptosis in cholangiocarcinoma cells by regulating the AGR2-mediated p38 MAPK pathway.

125I seeds can effectively inhibit the growth of a variety of cancer cells. It has been used in the treatment of a variety of cancers, and has achieved certain curative effect. However, to the best of our knowledge, no report has described the effects of 125I seeds on the biological functions of cholangiocarcinoma (CCA) and the mechanisms underlying the effects of the seeds on this cancer. In this study, we demonstrated that 125I seeds could inhibit the proliferation, migration and invasion of CCA cells, as well as promoting apoptosis and blocking the cell cycle in these cells. Moreover, 125I seeds inhibited the growth of CCA xenografts and promoted the apoptosis of CCA cells in vivo. Furthermore, transcriptome sequencing showed that 125I seeds could inhibit the growth of CCA by inhibiting the expression of AGR2 and regulating p38 MAPK pathway. Finally, this finding indicated that 125I seeds can inhibit proliferation and promote apoptosis in CCA cells by inhibiting the expression of AGR2 and DUSP1 and increasing the expression of p-p38 MAPK and p-p53. This study provides a new research direction for studies investigating the mechanisms underlying the effects of 125I seeds on CCA.

Origins of selective differential oxidation of ß-lactam antibiotics with different structure in an efficient visible-light driving mesoporous g-C3N4 activated persulfate synergistic mechanism.

In the study, an efficient '1 + 1 > 2' synergistic coupling system driven by visible light consisting of mesoporous g-C3N4 (MCN) and persulfate (PS)was constructed. The free radical transformation, electron transfer and non-covalent interaction between the MCN layer and PS in the system were explored via experiments and DFT calculations. The similarity for the fate of the seven ß-lactam antibiotics with typical structures in the oxidation system was studied systematically in depth. First, the consistencies and differences of the seven antibiotics were summarized from three aspects: three-dimensional structures, electron cloud distributions, and the vulnerable sites. Notably, the selective differential degradation of ß-lactam antibiotics in the MCN/PS system was speculated to be related with the molecular ionization potential (MIP), as a key index to describe the difficulty of oxidation. The distribution relationship between MIP and the oxidation kinetic constant (K) was explored and showed the following trend: a higher MIP indicates a weaker ability to provide electrons, and this leads to a greater resistance to oxidative degradation. In total, four main oxidation pathways of ß-lactam antibiotics were systematically summarized combining HPLC-QTOF-MS and the simplified Fukui function calculation. The toxicity assessment of intermediate products provided by the T.E.S.T software of USEPA also shows a decreasing trend in the oxidation process. In the end, the superior practicability and stability of the MCN/PS system was verified by complex environment simulation and cyclic test. This research clarified the selective differential degradation mechanism of ß-lactam antibiotics and provided a possible idea for the effective removal of refractory organic pollutants in water.

Effect of carbon nitride synthesized by different modification strategies on the performance of carbon nitride/PVDF photocatalytic composite membranes.

Carbon nitride (CN)/polyvinylidene fluoride (PVDF) photocatalytic composite membrane (PCM) is considered as a promising candidate to improve the anti-fouling characteristic of conventional PVDF membrane and overcome the difficulty encountered during recovery of powder catalyst simultaneously. However, the effects of differently-modified CN on PCM and its mechanism are still unclear. In this study, bulk-CN (BCN), carbon defects CN (CCN), nitrogen defect CN (DCN), mesoporous CN (MCN), and nitrogen-rich CN (NCN) were incorporated into PVDF by phase inversion method. The influence of changes in the physical and chemical properties of CN, including hydrophilic groups, photocatalytic activity, and particle size, on the permeability, anti-fouling characteristic, and photocatalytic self-cleaning activity of CN/PVDF was systematically analyzed. The mechanism of excellent performance of PCM was revealed by experimental test and theoretical calculation. The flux of PCM was significantly improved by increasing the hydrophilic group on modified CN. However, the differences in particle size and interaction between different types of modified CN and PVDF chains endowed the CN/PVDF with different porosity. DCN/PVDF showed high porosity and hydrophilicity, leading to high water flux and rejection rate of 293.6 L (m2 h)-1 and 90.1%, respectively. Compared to pure PVDF, the flux recovery rate of DCN30/PVDF increased by 27.6%, and the irreversible fouling decreased from 36.9% to 9.2%. The modified CN/PVDF showed excellent photocatalytic activity for the removal of cefotaxime (CFX) and E. coli. Owing to the narrow band gap of DCN, large specific surface area, and low photogenerated carrier recombination rate, the CFX removal rate reached 99% in 2 h, and E. coli inactivation achieved 3.7 log within 4 h via DCN30/PVDF.

Screening Prognosis-Related lncRNAs Based on WGCNA to Establish a New Risk Score for Predicting Prognosis in Patients with Hepatocellular Carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) remains an important cause of cancer death. The molecular mechanism of hepatocarcinogenesis and prognostic factors of HCC have not been completely uncovered. METHODS: In this study, we screened out differentially expressed lncRNAs (DE lncRNAs), miRNAs (DE miRNAs), and mRNAs (DE mRNAs) by comparing the gene expression of HCC and normal tissue in The Cancer Genome Atlas (TCGA) database. DE mRNAs were used to perform Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Then, the miRNA and lncRNA/mRNA modules that were most closely related to the survival time of patients with HCC were screened to construct a competitive endogenous RNA (ceRNA) network by weighted gene coexpression network analysis (WGCNA). Moreover, univariable Cox regression and Kaplan-Meier curve analyses of DE lncRNAs and DE mRNAs were conducted. Finally, the lasso-penalized Cox regression analysis and nomogram model were used to establish a new risk scoring system and predict the prognosis of patients with liver cancer. The expression of survival-related DE lncRNAs was verified by qRT-PCR. RESULTS: A total of 1896 DEmRNAs, 330 DElncRNAs, and 76 DEmiRNAs were identified in HCC and normal tissue samples. Then, the turquoise miRNA and turquoise lncRNA/mRNA modules that were most closely related to the survival time of patients with HCC were screened to construct a ceRNA network by WGCNA. In this ceRNA network, there were 566 lncRNA-miRNA-mRNA regulatory pairs, including 30 upregulated lncRNAs, 16 downregulated miRNAs, and 75 upregulated mRNAs. Moreover, we screened out 19 lncRNAs and 14 hub mRNAs related to prognosis from this ceRNA network by univariable Cox regression and Kaplan-Meier curve analyses. Finally, a new risk scoring system was established by selecting the optimal risk lncRNAs from the 19 prognosis-related lncRNAs through lasso-penalized Cox regression analysis. In addition, we established a nomogram model consisting of independent prognostic factors to predict the survival rate of HCC patients. Finally, the correlation between the risk score and immune cell infiltration and gene set enrichment analysis were determined. CONCLUSIONS: In conclusion, the results may provide potential biomarkers or therapeutic targets for HCC and the establishment of the new risk scoring system and nomogram model provides the new perspective for predicting the prognosis of HCC.

Matrine treatment induced an A2 astrocyte phenotype and protected the blood-brain barrier in CNS autoimmunity.

Type 1 astrocytes (A1), which are highly proinflammatory and neurotoxic, are prevalent in multiple sclerosis (MS). In addition, in MS and its animal model, experimental autoimmune encephalomyelitis (EAE), immune cells must cross the blood-brain barrier (BBB) and infiltrate into the parenchyma of the central nervous system (CNS) in order to induce neurological deficits. We have previously reported that treatment of EAE with matrine (MAT), a quinazine alkaloid derived from Sophorae Flavescens, effectively inhibited CNS inflammation and promoted neuroregeneration. However, the impact of MAT treatment on astrocyte phenotype is not known. In the present study, we showed that MAT treatment inhibited the generation of neurotoxic A1 astrocytes and promoted neuroprotective A2 astrocytes in the CNS of EAE, most likely by inhibiting production of the A1-inducing cytokine cocktail. MAT also downregulated the expression of vascular endothelial growth factor-A (VEGF-A) and upregulated tight junction proteins Claudin 5 and Occludin, thus protecting the BBB from CNS inflammation-induced damage. Moreover, MAT treatment promotes the formation of astrocyte tight junctions at glia limitans, thereby limiting parenchymal invasion of the CNS by immune cells. Taken together, the inhibition of A1 astrogliogenesis, and the dual effects on the BBB and astrocytic glia limitans, may be the mechanisms whereby MAT significantly improves EAE clinical scores and neuroprotection.

Illumination of Molecular Pathways in Multiple Sclerosis Lesions and the Immune Mechanism of Matrine Treatment in EAE, a Mouse Model of MS.

The etiology of multiple sclerosis (MS) is not clear, and the treatment of MS presents a great challenge. This study aimed to investigate the pathogenesis and potential therapeutic targets of MS and to define target genes of matrine, a quinolizidine alkaloid component derived from the root of Sophorae flavescens that effectively suppressed experimental autoimmune encephalomyelitis (EAE), an animal model of MS. To this end, the GSE108000 gene data set in the Gene Expression Omnibus Database, which included 7 chronic active MS lesions and 10 control samples of white matter, was analyzed for differentially expressed genes (DEGs). X cell was used to analyze the microenvironmental differences in brain tissue samples of MS patients, including 64 types of immune cells and stromal cells. The biological functions and enriched signaling pathways of DEGs were analyzed by multiple approaches, including GO, KEGG, GSEA, and GSVA. The results by X cell showed significantly increased numbers of immune cell populations in the MS lesions, with decreased erythrocytes, megakaryocytes, adipocytes, keratinocytes, endothelial cells, Th1 cells and Tregs. In GSE108000, there were 637 DEGs, including 428 up-regulated and 209 down-regulated genes. Potential target genes of matrine were then predicted by the network pharmacology method of Traditional Chinese medicine, and 12 key genes were obtained by cross analysis of the target genes of matrine and DEGs in MS lesions. Finally, we confirmed by RT-PCR the predicted expression of these genes in brain tissues of matrine-treated EAE mice. Among these genes, 2 were significantly downregulated and 6 upregulated by matrine treatment, and the significance of this gene regulation was further investigated. In conclusion, our study defined several possible matrine target genes, which can be further elucidated as mechanism(s) of matrine action, and novel targets in the treatment of MS.

Curcumin inhibits the growth of triple-negative breast cancer cells by silencing EZH2 and restoring DLC1 expression.

Enhancer of zeste homolog 2 (EZH2), an oncogene, is a commonly up-regulated epigenetic factor in human cancer. Hepatocellular carcinoma deletion gene 1 (DLC1) is an antioncogene that is either expressed at low levels or not expressed in many malignant tumours. Curcumin is a promising anticancer drug that has antitumour effects in many tumours, but its mechanism of action is unclear. Our research demonstrated that EZH2 was up-regulated in breast cancer (BC) tissues and cells, whereas DLC1 was down-regulated, and the expression of EZH2 and DLC1 was negatively correlated in BC. By analysing the characteristics of clinical cases, we found that positive expression of EZH2 and negative expression of DLC1 may be predictors of poor prognosis in patients with triple-negative breast cancer (TNBC). Moreover, knockdown of EZH2 expression restored the expression of DLC1 and inhibited the migration, invasion and proliferation, promoted the apoptosis, and blocked the cell cycle of MDA-MB-231 cells. Furthermore, we found that curcumin restored the expression of DLC1 by inhibiting EZH2; it also inhibited the migration, invasion and proliferation of MDA-MB-231 cells, promoted their apoptosis and blocked the cell cycle. Finally, xenograft tumour models were used to demonstrate that curcumin restored DLC1 expression by inhibiting EZH2 and also inhibited the growth and promoted the apoptosis of TNBC cells. In conclusion, our results suggest that curcumin can inhibit the migration, invasion and proliferation, promote the apoptosis, block the cycle of TNBC cells and restore the expression of DLC1 by inhibiting the expression of EZH2.

Anthocyanidin Extract from Summer-black-grape Affects the Expression of Ki-67 in Testis, Ovary of D-Galactose-induced Aging Mice.

The aim of this work was to analyze the expression of Ki-67 in ovary, testis of aging mice with anthocyanidin extract from Summer-black-grape. ICR mice (the aging group and the anthocyanidin-groups (50 mg/kg/D-group, 75 mg/kg/D-group and 100 mg/kg/D-group) were employed to evaluate the effect of grape anthocyanidin on reproductive system. The results showed that the anthocyanidin had strong scavenging ability for free radicals, the level of oxidation in serum of mice treated with anthocyanidin was low, and the pathological changes were not obvious. In the anthocyanidin group, the Ki-67 positive particles in the testis and ovary tissue were significantly decreased. The anthocyanidin of Summer-black-grape reduces the expression of Ki-67 protein in the testis or ovary of aging mice. In gonadal cells of aging mice, the anthocyanidin were shown protective effect in the proliferation.