Association between BRAF expression in circulating tumor cells and the clinical feature of patients with multiple myeloma.

Multiple myeloma (MM) is the most common hematological malignancy with uncontrolled proliferation of monoclonal plasma cells. Despite treatment improvements, MM remains an incurable disease for most patients. Therefore, promising molecular markers are required for MM treatment decisions. In the present study, we explored the relationship between the BRAF expression in circulating tumor cells (CTCs) and the clinical features of patients with MM. The results showed that CTCs were associated with MM staging, and the expression of BRAF was associated with different CTCs. Moreover, the BRAF gene was correlated with patients' white blood cells, blood albumin levels, and Eastern Cooperative Oncology Group (ECOG) score. BRAF expression positively correlated with total CTCs, hybrid CTCs, and mesenchymal CTCs. Taken together, CTCs tightly correlated with the clinical stages and characteristics of MM. Our findings may provide a promising prognosis biomarker for MM treatment decisions.

Artemisinin protects endothelial function and vasodilation from oxidative damage via activation of PI3K/Akt/eNOS pathway.

INTRODUCTION: Previous studies showed that artemisinin (ART) may be useful in the protection against the early development of atherosclerosis, but the effects of ART on vasodilation and eNOS remained unclear. OBJECTIVES AND METHODS: In the current study, we investigated the protective effect of ART on endothelial cell injury induced by oxidative stress and its underlying mechanism via MTT assay, Flow Cytometry Assay, Vasodilation study, Western blotting and vivo assay. RESULTS: We found that pretreatment of human umbilical vein endothelial cells (HUVECs) with ART significantly suppressed H2O2-induced cell death by decreasing the extent of oxidation and MDA activity, activating SOD, increasing NO production and inhibiting caspase 3/7 activity. Meanwhile, we also found that ART was able to activate PI3K/Akt/eNOS pathway. PI3K inhibitor LY294002 or Akt kinase specific inhibitor Akt inhibitor VIII blocked the protective effect of ART. To explore the effect of ART in the damage of vasodilation induced by H2O2 in mice, we treated the aortic ring from C57BL/6 mice with H2O2 with or without ART, the results demonstrated that ART ameliorated endothelium-dependent vasodilation damage induced by H2O2. CONCLUSION: Taken together, these data suggest that ART is able to protect endothelial function and vasodilation from oxidative damage, at least in part through activation of PI3K/Akt/eNOS pathway. Our findings indicate that artemisinin maybe as a potential therapeutic agent for patients with atherosclerosis.

Exosomal mRNAs and lncRNAs involved in multiple myeloma resistance to bortezomib.

The bone marrow microenvironment plays an essential role in multiple myeloma (MM) progression. We aimed to explore the alterations of levels of long noncoding RNAs and messenger RNAs (mRNAs), derived from exosomes in peripheral blood, in resistance to bortezomib (Btz) of MM patients. Peripheral blood samples were collected from five Btz-resistant and five Btz-sensitive MM patients. Exosomes in patients' peripheral blood were enriched, and the profiles of long noncoding RNAs (lncRNAs) and mRNAs in exosomes were determined using deep sequencing. Bioinformatics analysis was performed to explore biological function. MTS was employed to determine the viability of Roswell Park Memorial Institute (RPMI) 8226 and LP-1 cells incubated with exosomes derived from Btz-resistant patients. Quantitative polymerase chain reaction (qPCR) was used to evaluate the levels of exosomal FFAR1, SP9, HIST1H2BG, and ITIH2. Incubation with Btz-resistant patient-derived exosomes significantly increased the viability of Btz-treated RPMI 8226 and LP-1 cells in a dose-dependent manner. We identified 482 lncRNAs and 2099 mRNAs deregulated in exosomes of the Btz-resistance group; and 78 mRNAs were enriched in DR-related pathways, including mammalian target of rapamycin, platinum drug resistance, and the cAMP and phosphoinositide 3-kinase-Akt signaling pathways. qPCR results verified the increases in FFAR1 and SP9 and decreases in HIST1H2BG and ITIH2 in Btz-resistant patient-derived exosomes. Moreover, exosomal FFAR1 and SP9 exhibited potential as independent prognostic indicators of survival of MM patients. Our study reveals significant dysregulation of exosomal RNA components in the Btz-resistant group of MM patients as well as several mRNAs that may be used as biomarkers of prognosis of MM patients that are resistant to Btz.

[Peripheral blood exosomes from patients with multiple myeloma mediate bortezomib resistance in cultured multiple myeloma cells].

OBJECTIVE: To investigate the role of exosome in mediating bortezomib (Btz) resistance in multiple myeloma cells in vitro and explore the underlying mechanisms. METHODS: Peripheral blood samples were collected from 15 patients with multiple myeloma with Btz tolerance, and serum exosomes were isolated by ultracentrifugation and identified with electron microscopy, NTA and Western blotting. In vitro cultured multiple myeloma cells were treated with gradient concentrations of Btz to determine the optimal drug concentration for subsequent experiment. The cells were pretreated with different concentrations of exosomes, and their sensitivity to BTZ was assessed using MTS assay. We searched the exosome database Exocarta and used STRING to generate the network map and the protein interaction graph. RESULTS: The diameters of the vesicles isolated from the peripheral blood of the patients were mostly below 200 nm with a mean particle size of 153 nm and a mode of 140.1 nm. The results of Western blotting showed that the isolated exosomes expressed the marker proteins CD63, Tsg101 and Alix. In cultured multiple myeloma cells, pretreatment with exosomes resulted in a decreased sensitivity of the cells to bortezomib, and longer treatment durations and higher exosome concentrations consistently enhanced the resistance of the cells to the same Btz concentration. Analysis of the Exocarta database identified human serum exosomal proteins ABCB1, ABCB4, PDCD6IP, and EGFR, among which EGFR served as a network node. CONCLUSIONS: Exosome within a specific concentration range may serve as a signal carrier to mediate the resistance of multiple myeloma cells to Btz. EGFR likely plays a key role to promote exosome-mediated Btz resistance in myeloma cells.

Meta-analysis of reference values of haemostatic markers during pregnancy and childbirth.

Previously reported haemostatic reference intervals in normal pregnancy displayed considerable contradictions to establish convince gestational age-related haemostatic reference values. 30 clinical reports were recruited to collect and assemble existing clinical reports from the database D-dimer levels increased progressively with gestational ages and exceeded conventional value of 1 mg/L after 29-36 weeks, and reached a peak at 24 h postpartum with mean value of 6.44 mg/L [95% confidence interval (CI): 5.84 to 7.05] and returned to 0.79 mg/L (95% CI: 0.43 to 1.16) at 1-8 weeks postpartum. Analogously, the level of fibrinogen gradually increased throughout the pregnancy, and peaked at 48-72 h after birth, with mean value of 9.05 g/L (95% CI: 2.22 to 15.89) and then returned to 3.62 g/L (95% CI: 3.03 to 4.20) at 1-8 weeks postpartum. However, in the middle trimester, asynchronously prothromb in fragments 1 + 2 (F1+2) level elevated and reached a peak at 28-36 weeks with mean value of 3.05 nmol/L (95% CI: 2.41 to 3.70), and then decreased in the later trimester, and reached 1.92 nmol/L (95% CI: 0.58 to 3.27) at 48-72 h post-partum, close to normal levels. Previously reported gestational age-related haemostatic reference intervals in pregnancy could not be used as a standard.