Ethanol extract of Eclipta prostrata induces multiple myeloma ferroptosis via Keap1/Nrf2/HO-1 axis.

BACKGROUND: Multiple myeloma (MM) is an incurable hematological malignancy with limited therapeutic efficacy. Eclipta prostrata is a traditional Chinese medicinal plant reported to possess antitumor properties. However, the effects of E. prostrata in MM have not been explored. PURPOSE: The aim of this study was to define the mechanism of the ethanol extract of E. prostrata (EEEP) in treating MM and identify its major components. METHODS: The pro-ferroptotic effects of EEEP on cell death, cell proliferation, iron accumulation, lipid peroxidation, and mitochondrial morphology were determined in RPMI-8226 and U266 cells. The expression levels of nuclear factor erythroid 2-related factor 2 (Nrf2), kelch-like ECH-associated protein 1 (Keap1), heme oxygenase-1 (HO-1), glutathione peroxidase 4 (GPX4), and 4-hydroxynonenal (4HNE) were detected using western blotting during EEEP-mediated ferroptosis regulation. The RPMI-8226 and U266 xenograft mouse models were used to explore the in vivo anticancer effects of EEEP. Finally, high performance liquid chromatography (HPLC) and ultra-high-performance liquid chromatography-quadrupole/time-of-flight mass spectrometry system (UPLC-Q/TOF-MS) were used to identify the major constituents of EEEP. RESULTS: EEEP inhibited MM cell growth and induced cell death in vitro and in vivo. By promoting malondialdehyde and Fe2+ accumulation, lipid peroxidation, and GSH suppression, EEEP triggers ferroptosis in MM. Mechanistically, EEEP regulates the Keap1/Nrf2/HO-1 axis and stimulates ferroptosis. EEEP-induced lipid peroxidation and malondialdehyde accumulation were blocked by the Nrf2 activator NK-252. In addition, HPLC and UPLC-Q/TOF-MS analysis elucidated the main components of EEEP, including demethylwedelolactone, wedelolactone, chlorogenic acid and apigenin, which may play important roles in the anti-tumor function of EEEP. CONCLUSION: In summary, EEEP exerts its anti-MM function by inducing MM cell death and inhibiting tumor growth in mice. We also showed that EEEP can induce lipid peroxidation and accumulation of ferrous irons in MM cells both in vivo and in vitro, leading to ferroptosis. In addition, this anti-tumor function may be achieved by the EEEP activation of Keap1/Nrf2/HO-1 axis. This is the first study to reveal that EEEP exerts anti-MM activity through the Keap1/Nrf2/HO-1-dependent ferroptosis regulatory axis, making it a promising candidate for MM treatment.

Correlation Analysis of Umbilical Cord Blood Metabolic Phenotype and Inflammation in Patients with Gestational Diabetes Mellitus Complicated with Overweight and Obesity.

Background: Gestational diabetes mellitus (GDM) is a common metabolic disorder in pregnancy. The incidence rate is increasing year by year, which seriously threatens the safety of maternal and infant. Obesity is a vital factor in inducing GDM. Pregnant women with GDM account for a large proportion of overweight and obese pregnant women. Our study aimed to explore the potential mechanism of differential metabolites on inflammation and find the intervention and management methods for GDM in overweight and obese pregnant women. Methods: Umbilical cord blood samples and placenta were collected from normal weight pregnant women with GDM (control group) and overweight and obese pregnant women with GDM (obesity group) for a comparative study. Serum inflammatory factors IL-10, TNF-α, IL-6, lipopolysaccharide (LPS), and TLR4 expression were detected by ELISA. The expression levels of BCL-2 and caspase-3 were measured by Western blot. TUNEL staining was used to observe the apoptosis of placental villi. KEGG combined with metabolomics was used to compare the differences of metabolic maps between the two groups. Results: Compared with the control group, the level of anti-inflammatory factor IL-10 in the cord blood was decreased in the obesity group, while the levels of proinflammatory factors TNF-α, IL-6, and LPS were increased. In the placental tissues, the obesity group had higher concentrations of LPS, TLR4, and caspase-3 and lower concentration of BCL-2. Placental villi in the obesity group were more likely to undergo apoptosis than the control group. Correlation analysis showed that the above metabolite concentrations were negatively correlated with TNF-α or LPS. Conclusion: Metabolites could control obesity in the process of controlling the occurrence and development of inflammation.

Decoy engineering of the receptor-like cytoplasmic kinase StPBS1 to defend against virus infection in potato.

Potato virus Y (PVY) is an important pathogen of potato (Solanum tuberosum). Although the PBS1-RPS5 immune system is well documented in Arabidopsis thaliana, it has not been reported in potato. In Arabidopsis, the bacterial effector AvrPphB cleaves AtPBS1 to trigger an immune response. Here, we show that the AvrPphB-triggered immune response is mediated by StPBS1, a close homologue of AtPBS1 in potato. However, downstream signalling of StPBS1 was mediated by unknown resistance (R) proteins other than potato orthologues of AtRPS5 and HvPBR1, which is important for HvPBS1 signalling in barley. Immune signalling of StPBS1 is mediated by the AvrPphB C-terminal cleavage domain and an STKPQ motif, in contrast to AtPBS1-mediated immunity in which both AvrPphB cleavage fragments and an SEMPH motif are essential. The cleavage sequence of AvrPphB in StPBS1 was replaced with that of the PVY NIa-Pro protease to obtain StPBS1NIa . StPBS1NIa overexpression potato displayed stronger immunity to PVY infection than did the StPBS1 transgenic lines. StPBS1NIa was cleaved at the expected target site by NIa-Pro protease from PVY. Thus, we characterized the function of StPBS1 in potato immunity and provide a biotechnology control method for PVY via transformation of decoy-engineered StPBS1NIa .

[Dendritic cell tumor vaccine assisted by Tiaohengfang polysaccharides enhances immune function of S180 tumor-bearing mice and inhibits tumor growth].

Objective To investigate the immunotherapeutic effect and mechanism of dendritic cell (DC) vaccine assisted by Tiaohengfang polysaccharides (ThPP) in S180 tumor-bearing mice. Methods Mouse bone marrow-derived cells were cultured in vitro and mature DCs were obtained with the assistance of cytokines and ThPP. The expression of CD80 and CD86 of DCs induced by ThPP was examined, and S180 tumor cells were used as antigens to stimulate dendritic cells to become dendritic cell tumor vaccine. Tumor-bearing models were established in mice by S180 tumor cells inoculated into the armpit of the left forelimb, and the mice were randomly divided into four groups according to body mass, namely tumor-bearing blank group, positive control group (cyclophosphamide), dendritic cell vaccine group adjuvanted by ThPP and TNF-α. The tumor-bearing mice were treated on the 5th and 10th days after inoculation of tumor cells. The tumor-bearing mice were killed on the 12th day and the tumor inhibition was observed by the tumor mass detection. At the same time, peritoneal macrophages were isolated and cultured, and the expression of CD11b and IL-12 were measured by immunohistochemistry. The levels of serum IL-12 and TNF-α in the mice were detected by ELISA. The survival time of the other four groups of tumor-bearing mice was observed after treatment with the same method. Results The expression of CD80 and CD86 in the TNF-α group and ThPP group were higher than those in the blank control group, and the ThPP group was more significant. The tumor inhibition rate and survival extension period of ThPP, TNF-α and positive groups were significantly higher than those of the model blank group. The levels of serum IL-12 and TNF-α in the ThPP group were higher than those in the positive cyclophosphamide group and model black group. There was no significant difference between the ThPP group and TNF-α group. The expression of CD11b in the macrophages of ThPP group was lower than that in the model blank group and positive group, while the expression of IL-12 in the macrophages of ThPP group was higher than that in the model blank group and positive group, without significant difference compared with TNF-α group. Conclusion ThPP-adjuvanted DC tumor vaccine can inhibit tumor growth and prolong survival time of S180 tumor-bearing mice, which is related to promoting the maturation of DCs and increasing the secretion of IL-12 and TNF-α.

Selenium Levels in Community Dwellers with Type 2 Diabetes Mellitus.

Selenium (Se) is an essential micronutrient required by the human body and exerts biological functions through selenoproteins. The relationship between Se and diabetes remains ambiguous. This study aimed at, through measurements of serum selenoprotein P (SelP), glutathione peroxidase (GPx3), and Se content in serum and hair, evaluating Se levels in community dwellers with type 2 diabetes mellitus (T2DM). A total of 336 subjects, including 176 T2DM patients (65.5 ± 8.7 years old) and 160 healthy residents (63.7 ± 9.8 years old, as controls), were recruited in this cross-sectional community-based study performed in Suzhou. Samples of fasting venous blood and hair were collected for measurements of Se levels, glycometabolism, and biochemistry parameters. We found that the serum Se level and SelP concentration in T2DM patients were substantially higher than those in healthy residents (p < 0.05). There was no significant difference in GPx3 activity and hair Se level between the two groups of subjects (p > 0.05). Serum Se was positively correlated with both GPx3 and hair Se in T2DM patients (r = 0.167 and 0.164, respectively, p < 0.05) and negatively correlated with SelP in healthy controls (r = - 0.293, p < 0.05). In conclusion, this study showed significantly higher levels of serum Se and SelP in community dwellers with T2DM than in matched healthy residents.

Traditional Chinese Medicine Extract from Huaier Increases the Expression of Duffy Antigen Receptor for Chemokines and Reduces the Expression of Its Ligands.

AIMS: The aim of the present study is to investigate whether the aqueous extract from Huaier, a traditional Chinese medicine (TCM), can affect the expression of Duffy antigen receptor for chemokines (DARC) and its ligands. Moreover, we compare the status of DARC in primary and metastatic breast cancer tissues from the same patient. METHODS: Immunohistochemistry was used to detect the expression of DARC in primary and metastatic focuses in 30 patients with breast cancer. The effect of Huaier aqueous extract on the expression of DARC and its ligands was investigated by quantitative real-time polymerase chain reaction, Western blotting, and enzyme-linked immunosorbent assay. RESULTS: The expression score of DARC in primary focuses was significantly higher than that in metastatic focuses, while changes of ER, PR, and HER2 receptors were not significantly different between primary and metastatic focuses. Huaier aqueous extract promoted the expression of DARC and reduced the secretion of CC chemokine ligand 2 (CCL-2), CXC chemokine ligand 8 (CXCL-8, IL-8), matrix metalloproteinase 2 (MMP-2), and CXC chemokine ligand 1 (CXCL-1). CONCLUSION: The present study demonstrates that difference in expression level of DARC between primary and metastatic focuses of breast cancer was significant, while differences in expression of ER, PR, and HER2 between primary and metastatic focuses were not significant. DARC may play a negative role in the metastasis of breast cancer. Traditional Chinese medicine extract from Huaier can increase DARC expression and reduce the expression of its ligands such as CCL-2, IL-8, MMP-2, and CXCL-1.

[Expression of YY 1 protein in human insulinoma and its clinical implication].

OBJECTIVE: To investigate the expression of Yin Yang 1 (YY1) protein in human insulinoma and explore its clinical significance. METHODS: Nineteen pancreatic neuroendocrine tumor tissue were collected from patients treated in Nanfang Hospital between 2000 and 2014. The protein expression of YY1 in benign and malignant insulinoma tissues were detected by immunohistochemistry. RESULTS: Positive expression for YY1 protein was detected in both benign and malignant tumor tissues, but the malignant tissues had a significantly greater intensity of YY1 expression than the benign tissues (P=0.042). The intensity of YY1 expression was positively correlated with the nature of the tumor, and the insulinomas with high expressions of YY1 had significantly greater malignant potentials (P=0.037). CONCLUSION: The high expression of YY1 protein is associated with the development of insulinima. YY1 may serve as a new tumor marker for detecting the malignant transformation of insulinoma.

Thiamine deficiency increases ß-secretase activity and accumulation of ß-amyloid peptides.

Thiamine pyrophosphate (TPP) and the activities of thiamine-dependent enzymes are reduced in Alzheimer's disease (AD) patients. In this study, we analyzed the relationship between thiamine deficiency (TD) and amyloid precursor protein (APP) processing in both cellular and animal models of TD. In SH-SY5Y neuroblastoma cells overexpressing APP, TD promoted maturation of ß-site APP cleaving enzyme 1 (BACE1) and increased ß-secretase activity which resulted in elevated levels of ß-amyloid (Aß) as well as ß-secretase cleaved C-terminal fragment (ß-CTF). An inhibitor of ß-secretase efficiently reduced TD-induced up-regulation of Aß and ß-CTF. Importantly, thiamine supplementation reversed the TD-induced alterations. Furthermore, TD treatment caused a significant accumulation of reactive oxygen species (ROS); antioxidants suppressed ROS production and maturation of BACE1, as well as TD-induced Aß accumulation. On the other hand, exogenous Aß(1-40) enhanced TD-induced production of ROS. A study on mice indicated that TD also caused Aß accumulation in the brain, which was reversed by thiamine supplementation. Taken together, our study suggests that TD could enhance Aß generation by promoting ß-secretase activity, and the accumulation of Aß subsequently exacerbated TD-induced oxidative stress.

Signaling through Tyr985 of leptin receptor as an age/diet-dependent switch in the regulation of energy balance.

Leptin regulates energy homeostasis through central activation of multiple signaling pathways mediated by Ob-Rb, the long form of leptin receptor. Leptin resistance underlies the pathogenic development of obesity, which is closely associated with environmental factors. To further understand the physiological function of leptin signaling mechanisms, we generated a knock-in line of mice (Y985F) expressing a mutant Ob-Rb with a phenylalanine substitution for Tyr985, one of the three intracellular tyrosines that mediate leptin's signaling actions. Surprisingly, whereas young homozygous Y985F animals were slightly leaner, they exhibit adult-onset or diet-induced obesity. Importantly, both age-dependent and diet-induced deterioration of energy balance was paralleled with pronounced leptin resistance, which was largely attributable to attenuation of leptin-responsive hypothalamic STAT3 activation as well as prominently elevated expression of hypothalamic SOCS3, a key negative regulator of leptin signaling. Thus, these results unmask distinct binary roles for Try985-mediated signaling in energy metabolism, acting as an age/diet-dependent regulatory switch to counteract age-associated or diet-induced obesity.

Senescence accelerated mouse strain is sensitive to neurodegeneration induced by mild impairment of oxidative metabolism.

Neuronal loss and impairment of oxidative metabolism are frequently observed in aging associated neurodegenerative diseases. Thiamine deficiency (TD) induces the region selective neuronal loss in the brain, which has been used to model neurodegeneration, accompanied by mild impairment of oxidative metabolism. C57BL/6 mice were commonly used animals for TD experiments; however, the individual variations among C57BL/6 mice in response to TD limited the consistence of brain pathology. The senescence accelerated prone 8 (SAMP8) mouse strain exhibits age-related morphological changes in the brain and deficits in learning and memory. In this study, we compared the effects of TD on SAMP8 mice, senescence accelerated resistant 1 (SAMR1) mice and C57BL/6 mice. TD-induced body weight loss in SAMP8 mice was much greater than in SAMR1 and C57BL/6 mice. In addition, earlier and more severe loss of neurons in the submedial thalamic nucleus (SmTN) of the thalamus was detected in the SAMP8 mice. After 8 days of TD (TD8), the loss of NeuN-positive neurons in the SmTN of SAMP8, SAMR1 and C57BL/6 mice was 65%, 50%, and 36%, respectively. TD also caused accumulation of amyloid precursor protein (APP) in the thalamus. After TD10, APP immunoreactivity in the thalamus of SAMP8 was much more intense than that of SAMR1 and C57BL/6 mice. These results suggest that SAMP8 mice are sensitive to TD and therefore offer a useful model for studying aging related neurodegeneration caused by the impairment of oxidative metabolism.

[Study on separation and purification of total flavones from Smilax china by macroporous absorption resin].

OBJECTIVE: To investigate the technological parameters of the purification process of total flavones from Smilax china with macroporous absorption resin. METHOD: The technical process for purification of total flavones with the optimum macroporous absorption resin was screened by yield of total flavones product. RESULT: The D140 macroporous absorption resin had the best separating efficiency when the flavones content in the liquid was 0.5 g x mL(-1) equivalent to raw material, the volume of drug 18 BV (resin bad volume) with the adsorption-power 2 BV x h(-1), and the volume of 60% (mL x mL(-1)) ethanol as eluant 5-10 BV (resin bad volume) with desorption-power 1 BV x h(-1). The obtained flavones product has total flavones recovery rate of 84.72%. CONCLUSION: The treatment of regenerated resin is easy, this method is advisable.