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Additional file 1. Fig. S1: Patients were divided into five groups according to the IPSS-R scoring system, and a comparison of the differences between groups was performed for each immunological index. P < 0.05 was statistically significant. Fig. S2: In the other two scoring systems, WPSS and IPSS, the serum IL-4 levels in middle and high-risk MDS patients have a predictive prognostic value.

Supplementary Material 1

Purpose: To find bone marrow immune detection index which related to MM patients therapy. Method: We analyzed ninety-three newly diagnosed MM patients retrospectively and their the stage of prognosis, the clinical and induced-therapy response datasets. Result: In our study, Eight-color flow cytometry, a method commonly used to detect MM cell loads, was used to analyze seven bone marrow immune cell groups (B, NK, T, naive B, immature B, memory B, and plasmablast like cells). The percentage of B cells significantly decreased in stage III, compared with stage I patients, (P = 0.047, P = 0.038) according to the ISS and R-ISS. In the clinical characteristics, naive B cell numbers increased in the Hb(g/L)≥100 group, when compared with the Hb(g/L)< 100 group (P = 0.028). The immature B (P = 0.017), plasmablast like cell subgroups(P = 0.037) increased in MM patients, who achieved a very good partial response(VGPR), after four cycles of the new drug-based induced therapy. The ROC results indicated the combination of the seven immune subgroups had predictive values (AUC = 0.840; P < 0.0001). Conclusion: comprehensively analyze seven bone marrow immune cell subgroups can be a useful approach for predicting the induce therapy response in MM patients.

Additional file 1: Supplementary tables and figures.

Additional file 1: Figure S1. CD59 and FLAER expression levels in erythrocytes (A a healthy volunteer, B a PNH patient).

Bone marrow microenvironment plays an important role in the development of myelodysplastic syndrome (MDS). Iron overload (IO) may be related to the prognosis of the MDS patients. The effects of IO damaging mesenchymal stromal cells (MSCs) from MDS patients and promoting leukemia transformation were investigated. In our study, the IO group showed the decreased quantity and weakened abilities on proliferation and differentiation of MSCs. The gene expressions of VEGFA, CXCL12 and TGF-β1 in MSCs regulating hematopoiesis were significantly downregulated in IO group. The levels of reactive oxygen species (ROS) and apoptosis in IO group were significantly higher than those in non-iron overload (NIO) group, with highly expressed caspase3 and β-catenin. Meanwhile, the damages of MSCs caused by IO could be partially reversed by antioxidant or iron chelator. Due to the higher gene mutation frequency in IO group, the effects of IO promoting leukemia transformation were investigated. The IO MDS/AML group had a lower quantity of MSCs compared with IO MDS group, with significantly decreased expression of p-AKT. However, there was no significant differences in apoptosis, gene expressions of VEGF, CXCL12 supporting hematopoiesis, and β-catenin involving leukemia transformation in MSCs between them. In conclusion, IO damages MSCs of MDS patients by inducing ROS related apoptosis and activating caspase3 signals, which could be partially reversed by antioxidant or iron chelator. The increased genetic mutations and activated β-catenin signals in MSCs of MDS patients caused by IO may involve in promoting leukemia transformation. Disclosures No relevant conflicts of interest to declare.

Background: To investigate the correlation of adipose cytokines (visfatin, leptin, and adiponectin) with multiple myeloma bone disease (MBD) and explore the effects and mechanism of adiponectin on the differentiation and maturation of osteoclasts in multiple myeloma (MM). Methods: The levels of visfatin, leptin and adiponectin were measured .Their relationships between the index of myeloma tumor load and bone disease were analyzed. RT-PCR was used to detect the expressions of RANKL, OSCAR, TRAP and Cathepsin K genes. Flow cytometry (FCM) was used to detect the expressions of AdipoR1 and phosphorylation of mTOR pathway related proteins mTOR and 4EBP1. Results :There were no significant linear correlation between leptin, visfatin and the indexes of myeloma tumor load and bone disease. Serum adiponectin levels were significantly higher in newly diagnosed multiple myeloma patients and negatively linear correlated with β2-microglobulin.There is a positive linear correlation relationship between the adiponectin and osteocalcin. The number of maturation osteoclasts in adiponectin group was less than the control group. Adiponectin also inhabits the mRNA level of osteoclast related factor- RANKL, OSCAR, TRAP and Cathepsin K. Flow cytometry showed that adiponectin increased the expression of AdipoR1 on the surface of osteoclast precursor((26.21±4.27)% vs (29.86±6.23)%, p Conclusion: Adiponectin inhibits the differentiation and maturation of osteoclasts via increasing the expression of AdipoR1 and reducing the phosphorylation levels of mTOR and 4EBP1 proteins in MM patients. Disclosures No relevant conflicts of interest to declare.

In this study, we intend to detect the expression of TIM3 on peripheral blood NK cells in SAA patients to reveal the further immune pathogenesis of SAA. Furthermore, we tried to further elucidate the changes of functions of TIM3+ NK and TIM3-NK cells in SAA by measuring the functional molecules and cytotoxic activity of TIM3+ NK and TIM3-NK cells. Finally, we observed the therapeutic effects of TIM3 blocker, TIM3+ NK infusion and TIM3-NK infusion on SAA mice model. 1.The TIM3 expression on NK cells in SAA untreated patients was significantly lower than that in SAA remission patients (P 2. TIM3-NK cells expressed higher NKG2D and Granzyme B than TIM3+ NK cells in untreated SAA patients. The expression of NKG2A, CD158a and CD158b on TIM3-NK cells were lower than TIM3+ NK cells. 3. The expression of CD80 and CD86 were significantly decreased after being incubated with TIM3-NK and TIM3+NK cells in SAA, especially mDC+ TIM3-NK group, significantly lower than mDC+TIM3+NK group(P 4. The apoptosis rate (AR) of K562 cells were significantly increased after being incubated with TIM3-NK and TIM3+ NK cells in SAA, especially K562+TIM3-NK group, significantly higher than K562+TIM3+NK groups(P 5. There was no significant difference in the level of AKT of receptor post-signal pathway protein between TIM3-NK and TIM3+ NK cells in patients with SAA, but the level of P-AKT in TIM3-NK cells is higher than TIM3+ NK cells. 6. AA mice model was established. The TIM3 expression on peripheral blood NK cells in SAA mice was significantly lower than that in TBI mice (P 7. On the 17th day of model establishment, the weight, hemogram and bone marrow cells count of AA mice were significantly lower than that of NC group (p0.05), and combined with CsA has no significant synergistic effect. The therapeutic effect of TIM3-NK cell infusion group was better than that of TIM3+ NK cell infusion group. The therapeutic effect of CsA combined with TIM3-NK cell infusion group was more significant than that of CsA alone group. TIM3-NK cell infusion therapy may have some synergistic effect with CSA. Conclusions 1. In this study, we found that untreated patients with SAA had lower TIM3 expression on NK cells compared with normal controls, andwere correlated with the severity of pancytopenia of SAA. 2. We further confirmed that the expression of activation molecules on TIM3-NK cells was increased and the killing function was enhanced compared with TIM3+NK cells. In addition, TIM3-NK cells have enhanced inhibition of mDCs and K562 cells and play an immunomodulatory role in SAA. Therefore, TIM3 exerts its inhibitory effect on NK cells and participates in the immune pathogenesis of SAA. Low expression of TIM3 contributes to the enhancement of NK cell function, which in turn inhibits the immune activation state of SAA and improves the disease level. 3. The expression of TIM3 on NK cells of AA mice decreased, and the activity of TIM3-NK cells was stronger than that of TIM3+ NK cells, which was consistent with the decrease of TIM3 on NK cells of SAA patients and the strong activity of TIM3-NK cells. After TIM3-NK cell reinfusion, the general condition, blood cell count and bone marrow cell count of SAA mice were improved, and the combined treatment of CsA was more effective. It may further clarify the immune pathogenesis of SAA and provide a new treatment target to improve the efficacy of SAA treatment. Disclosures No relevant conflicts of interest to declare.

OBJECTIVE: To study the effects of multiple myeloma (MM)-derived exosomes on the number and function of CD4+T, CD8+T and regulatory T cells in healthy donors (HD) and MM patients. METHODS: MM cell-derived (OPM2, U266) exosomes were extracted by ultracentrifugation and kit method and co-cultured with CD4+T, CD8+T and CD4+CD25+CD127dimT cells (Treg) from HD and MM patients sorted by magnetic beads. Flow cytometry was used to detect the apoptosis rate of T cells, level of perforin and granzyme B in CD8+T. Cell viability was detected by CCK8 kit, and IL-10 and TGF-β levels were detected by ELISA. RESULTS: HD-CD4+T apoptotic rate was higher in the OPM2 group (6.24±1.24%) and U266 group (5.42±1.07%) than in the control group (4.37±0.96%). The HD-CD4+T viability in U266 group (85.45±6.09%) decreased compared with the control group. The apoptotic rate of HD-CD8+T was decreased in the OPM2 group (9.97±1.28%) and the U266 group (11.00±1.75%) compared with the control group (16.12±2.95%). HD-CD8+T cell viability was increased in the OPM2 group (112.63±3.88%) and the U266 group (111.70±3.62%) compared with the control group. Perforin of HD-CD8+T in U266 group (9.34±2.36%) was lower than the control group (12.76±3.31%); Perforin of MM-CD8+T in OPM2 group (6.48±1.06%) and U266 group (5.63±1.15%) decreased compared with the control group (10.55 ± 2.50%). The apoptosis rate of HD-Treg was lower in OPM2 group (15.33±3.87%) and U266 group (11.71±2.71%) than in control group (19.61±3.50%). The apoptosis rate of MM-Treg was higher in U266 group (37.29±6.54%) than the control group (29.95 ± 6.68%). The viability of HD-Treg in U266 group (173.48±34.99%) was higher than the control group. The viability of MM-Treg in OPM2 group (82.29±12.16%) was lower than the control group. The supernatant IL-10 from HD-Treg in OPM2 group (18.53±8.08 pg/ml) was higher than the control group (8.49±4.02 pg/ml). The supernatant TGF-βfrom MM-Treg was decreased in the OPM2 group (290.29±23.21pg/ml) and the U266 group (230.03±15.96pg/ml) compared with the control group (387.49±21.60pg/ml) (P Conclusion: MM-derived exosomes can promote apoptosis and inhibit proliferation of HD-CD4+T, inhibit apoptosis and promote proliferation but inhibit perforin secretion of HD-CD8+T, inhibit apoptosis and promote proliferation HD-Treg, and promotes IL-10 secretion. Disclosures No relevant conflicts of interest to declare.

The effect of electron irradiation on the structure and mechanical properties of polyethylene cable was studied using ESR, posistron annihilation and SEM methods. It was found that radiation crosslinking enhanced the low temperature impact strength, declined the second yielding effect for uniaxial drawing of polyethylene cable.