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Hematologic malignancies (HMs) mainly include acute and chronic leukemia, lymphoma, myeloma and other heterogeneous tumors that seriously threaten human life and health. The common effective treatments are radiotherapy, chemotherapy and hematopoietic stem cell transplantation (HSCT), which have limited options and are prone to tumor recurrence and (or) drug resistance. Metformin is the first-line drug for the treatment of type 2 diabetes (T2DM). Recently, studies identified the potential anti-cancer ability of metformin in both T2DM patients and patients that are non-diabetic. The latest epidemiological and preclinical studies suggested a potential benefit of metformin in the prevention and treatment of patients with HM. The mechanism may involve the activation of the adenosine monophosphate-activated protein kinase (AMPK) signaling pathway by metformin as well as other AMPK-independent pathways to exert anti-cancer properties. In addition, combining current conventional anti-cancer drugs with metformin may improve the efficacy and reduce adverse drug reactions. Therefore, metformin can also be used as an adjuvant therapeutic agent for HM. This paper highlights the anti-hyperglycemic effects and potential anti-cancer effects of metformin, and also compiles the in vitro and clinical trials of metformin as an anti-cancer and chemosensitizing agent for the treatment of HM. The need for future research on the use of metformin in the treatment of HM is indicated.

β- elemene extracted from Curcuma zedoaria rhizome (commonly known as white turmeric) is an effective anticancer agent. There are limited reports on the use of this agent against THP-1 cells employing a mechanistic study. Therefore, as a foremost aim of the present study, the cytotoxic effect of elemene and its mechanism will be elucidated. For this aim, the method adopted was to treat THP-1 cells in a dose- and time- dependent manner with elemene and the cytotoxicity to be evaluated. The mRNA expressions of a set of autophagy and apoptosis related genes will be analyzed by quantitative PCR. The findings indicate that the IC50 values for 24, 48, 72 and 96 h of treated THP-1 cells were 64.71, 42.19, 25.29 and 20.21 μg/mL respectively. The expressions of autophagy related genes such as Beclin-1, LC3II, ATG-5 and XBP-1 were upregulated. The expressions of anti-apoptotic Bcl2 was upregulated, whereas, the expression of pro-apoptotic Bax was downregulated. Interestingly, the expressions of Caspase-3 and Caspase-8 were upregulated. To summarize, autophagy might have occurred in endoplasmic reticulum and there might be a crosstalk with apoptosis which could be the rationale behind the cytotoxic effects of elemene on THP-1 cells. Therefore, β- elemene could be a potential therapeutic agent for leukemia.

Background Novel fusion genes such as ZNF384, have been identified in B-cell precursor acute lymphoblastic leukemia (BCP-ALL) in recent years. Patients harboring ZNF384 rearrangements have a distinctive immunophenotype with weak CD10 and aberrant CD13 and/or CD33 expression. Thus, ZNF384-rearranged ALL is a unique subtype of BCP-ALL. However, research on the prognostic significance of ZNF384 rearrangements has been limited to date, especially in adolescents. Case presentation We described a 17-year-old adolescent who was diagnosed with ALL and had renal involvement as the first manifestation, which was very rare in the existing studies. FISH analysis indicated a rearrangement of ZNF384 according to its probe. The patient had a typical characteristic immunophenotype of ZNF384 rearrangement, with CD10 negativity and CD13 and CD33 positivity. She had an unfavorable prognosis because she responded poorly to chemotherapy and developed a relapse shortly after reaching CR. Conclusion The importance of ZNF384 rearrangements in terms of prognosis remains unclear. We reported an adolescent who was diagnosed with ZNF384-rearranged ALL with renal involvement. She underwent different therapies, but her prognosis remained poor. Since ZNF384 rearrangements may act as a prognostic predictor in children or adolescents, early detection based on its characteristic immunophenotype is of great necessity.

Diffuse large B‑cell lymphoma (DLBCL) is a common subtype of non‑Hodgkin lymphoma, which is curable in the majority of patients treated with rituximab plus cyclophosphamide, doxorubicin, vincristine and prednisone (R‑CHOP) immunochemotherapy. However, the therapeutic mechanism of R‑CHOP has not been elucidated. The GSE32918 and GSE57611 datasets were retrieved from The Gene Expression Omnibus database. The differentially expressed genes (DEGs) associated with R‑CHOP therapy were identified using limma. Combined with prognostic information in GSE32918, DEGs found to be significantly associated with prognosis were selected using univariate Cox regression analysis and a risk prediction model was constructed. Based on this model, the samples in the training set (GSE32918) were divided into high and low risk score groups according to the median risk score. A total of 801 DEGs were identified between the R‑CHOP treated DLBCL and primary DLBCL samples, from this 116 prognosis‑associated genes were selected. Using Cox proportional hazards model, an optimal combination of 12 genes [including calcium/calmodulin dependent protein kinase I (CAMK1), hippocalcin like 4 (HPCAL4) and ephrin A5 (EFNA5)] was selected, and the sample risk score prediction model was constructed and validated. The DEGs between high risk score and low risk score groups were significantly enriched in functions associated with 'response to DNA damage stimulus', and pathways including 'cytokine‑cytokine receptor interaction' and 'cell cycle'. The optimal combination of the 12 genes, including CAMK1, HPCAL4 and EFNA5, was found to be useful in predicting the prognosis of patients with DLBCL after R‑CHOP treatment. Therefore, these genes may be affected by R‑CHOP in DLBCL.

Human mucin-1 (MUC1) has attracted considerable attention owing to its overexpression in diverse malignancies. Here, for the rapid and efficient detection of MUC1, we present a SERS-colorimetric dual-mode aptasensor, by integrating SERS probes with magnetic separation, which has several distinctive advantages. Using such a dual-mode aptasensor, the colorimetric functionality is distinguishable by the naked eye, providing a fast and straightforward screening ability for the detection of MUC1. Moreover, SERS-based detection greatly improves the detection sensitivity, reaching a limit of detection of 0.1 U/mL. In addition, the combination of SERS and colorimetric method holds the advantages of these two techniques and thereby increases the reliability and efficiency of MUC1 detection. On the one hand, the magnetic nanobeads functionalized with MUC1-specific aptamer were utilized as an efficient capturing substrate for separating MUC1 from biological complex medium. On the other hand, the gold-silver core-shell nanoparticles modified with Raman reporters and the complementary sequences of MUC1 were used as the signal indicator, which could simultaneously report the SERS signal and colorimetric change. This strategy can achieve a good detection range and realize MUC1 analysis in real patients' samples. Thus, we anticipate that this kind of aptasensor would provide promising potential applications in the diagnosis and prognosis of cancers. Graphical abstract.

HOX gene dysregulation is a common feature of acute myeloid leukemia (AML). The molecular mechanisms underlying aberrant HOX gene expression and associated AML pathogenesis remain unclear. The nuclear protein CCCTC-binding factor (CTCF), when bound to insulator sequences, constrains temporal HOX gene-expression patterns within confined chromatin domains for normal development. Here, we used targeted pooled CRISPR-Cas9-knockout library screening to interrogate the function of CTCF boundaries in the HOX gene loci. We discovered that the CTCF binding site located between HOXA7 and HOXA9 genes (CBS7/9) is critical for establishing and maintaining aberrant HOXA9-HOXA13 gene expression in AML. Disruption of the CBS7/9 boundary resulted in spreading of repressive H3K27me3 into the posterior active HOXA chromatin domain that subsequently impaired enhancer/promoter chromatin accessibility and disrupted ectopic long-range interactions among the posterior HOXA genes. Consistent with the role of the CBS7/9 boundary in HOXA locus chromatin organization, attenuation of the CBS7/9 boundary function reduced posterior HOXA gene expression and altered myeloid-specific transcriptome profiles important for pathogenesis of myeloid malignancies. Furthermore, heterozygous deletion of the CBS7/9 chromatin boundary in the HOXA locus reduced human leukemic blast burden and enhanced survival of transplanted AML cell xenograft and patient-derived xenograft mouse models. Thus, the CTCF boundary constrains the normal gene-expression program, as well as plays a role in maintaining the oncogenic transcription program for leukemic transformation. The CTCF boundaries may serve as novel therapeutic targets for the treatment of myeloid malignancies.

Photonic crystals are nanomaterials with unique structural coloration, and they have been used in multiple analyses, gene function analyses, and nanoarray biosensors. Here, we used agarose hydrogel microcarriers with characteristic reflection peak codes to improve the biocompatibility of the material and the efficiency of detection, which may be helpful for further clinical applications. The paper has two parts: condition optimization and product application. Goat antihuman immune globulin G, human immune globulin G, and fluorescein isothiocyanate-labeled goat antihuman immune globulin G are commonly used materials in double-antibody sandwich techniques, which are used for optimization in the application of advanced composite materials. Goat antihuman immune globulin G was immobilized on the surfaces of photonic crystal hydrogel beads through chemical bonding. When the target immune globulin G was captured, the fluorescence-labeled antibody could combine with photonic crystal hydrogel beads. The r...

Objectives The aim of this part is to analyze the efficacy of R-CHOP regimen in the treatment of diffuse large B-cell lymphoma (DLBCL), and to explore the risk factors of relapse and refractory DLBCL by analyzing the clinical characteristics of patients, and to study the predictive efficacy of these factors on the prognosis of patients. Methods Clinical data of 71 patients with de novo DLBCL from December 2012 to December 2018 in the Department of Hematology, Zhongda Hospital Affiliated to Southeast University were collected and retrospectively analyzed. The patients with DLBCL were divided into two groups according to who were refractory or relapse after initial therapy. Then the response rate and high-risk factors of refractory and relapse lymphoma were analyzed by the chi-square test and Mann-Whitney U test. Besides, Overall survival (OS) curves and prognosis factors were estimated by the Kaplan-Meier method, Log-rank test and Cox proportional hazard regression analysis. Furthermore, the patients were divided into two groups according to the IPI score, and a comparative analysis of survival rate was performed between subgroups. Results 1.There were 71 patients who were diagnosed with DLBCL, in which 45 cases achieved complete remission (CR), 11cases achieved partial remission (PR), and the total remission rate was 78.9%. By the end of the follow-up, 25 cases (35.2%) experienced refractory and relapse, and 17 cases (23.9%) died, with an average OS of 60 months and an average EFS of 52 months. 2.Univariate analysis showed that the B symptoms (P< 0.001), low levels of Hb (P< 0.001) and LMR (P< 0.001), high levels of NLR (P= 0.042), β2-MG (P= 0.011), hs-CRP (P= 0.002) and LDH (P= 0.017) were significantly related with refractory and relapse. Multivariate Logistic analysis showed B symptoms (P= 0.033) and high levels of β2-MG (P= 0.048) were independent risk factors for refractory and relapse lymphoma. 3.Kaplan-Meier method analysis showed that the OS of patients with either B symptoms (P< 0.001), low levels of Hb (P= 0.008) and LMR (P= 0.005), or high levels of β2-MG (P= 0.007), hs-CRP (P= 0.008) and LDH (P= 0.002) were significantly shorter than that of control group. Additionally, Cox regression methods analysis showed that B symptoms (P= 0.026) and high β2-MG (P= 0.038) were independent prognosis factors for DLBCL. 4.Kaplan-Meier analysis of survival between the low-risk IPI and high-risk IPI groups showed that the OS of patients with B symptoms in both IPI low-risk group (P = 0.013) and IPI high-risk group (P = 0.027) weresignificantly shorter than those without B symptoms. Moreover, there was no significant difference in the OS of patients with either low levels of Hb and LMR,or high levels of LDH, β2-MG, and hs-CRP in both group (P > 0.05). Conclusion B symptoms and high levels of β2-MG are independent risk factors for relapse and refractory, and are expected to be incorporated into the new prognostic score system. Part two: Clinical study of maintenance therapy for DLBCL Objectives The aim of this part was to explore the efficacy of different maintenance therapies on DLBCL, and evaluate the safety of rituximab and traditional Chinese medicine maintenance therapy. Methods Clinical data of 71 patients with de novo DLBCL from December 2012 to December 2018 in the Department of Hematology, Zhongda Hospital Affiliated to Southeast University were collected and retrospectively analyzed. Follow-up and study whether patients have undergone maintenance therapy and maintenance therapy regimen. Results Of the 71 patients with DLBCL, 11 cases received maintenance therapy after CR, of which 6 cases were maintained with rituximab and 5 cases maintained with traditional Chinese medicine (TCM). The median follow-up time was 27 months. By the end of the follow-up, none of the 11 cases had relapsed and no treatment-related adverse reactions occurred. In particular, 2 patients received autologous hematopoietic stem cell transplantation after achieving CR, and then took TCM for maintenance therapy. No tumor recurrence was seen during follow-up and the clinical indicators were normal and stable. Conclusion Rituximab and TCM have good efficacy and safety in the maintenance treatment of DLBCL. TCM maintenance treatment shows unique advantages, and it is expected to be widely used in clinic after further verification in the future. Key words Diffuse large B-cell lymphoma; maintenance therapy; rituximab; traditional Chinese medicine Disclosures No relevant conflicts of interest to declare.

Thrombotic diseases are major cause of cardiovascular diseases. This study was designed to investigate the effect of tachyplesin I on platelet aggregation and thrombosis. Platelet aggregation was analysed with a whole blood aggregometer. The mice were employed to investigate the effect of tachyplesin I on thrombosis in vivo. Tachyplesin I inhibited thrombin-induced platelet aggregation in a dose-dependent manner. Furthermore, tachyplesin I significantly reduced thrombosis in carrageenan-induced tail thrombosis model by intraperitoneal injection (0.1, 0.2 or 0.4 mg/kg) or intragastric administration (15, 30 or 60 mg/kg). Tachyplesin I also prolonged the bleeding time (BT) and clotting time (CT). The results revealed that tachyplesin I inhibited platelet aggregation and thrombosis by interfering the PI3K/AKT pathway. Tachyplesin I did not show significantly toxicity to mice under 300 mg/kg via intravenous injection. The results show that tachyplesin I inhibits thrombosis and has low toxicity. It is suggested that tachyplesin I has the potential to develop a new anti-thrombotic drug.

Objective: To explore the non-genetic prognostic factors in patients with multiple myeloma (MM). Methods: Kaplan-Meier survival curve, Log-rank test and Cox regression model were used to carry out univariate and multivariate analysis on clinical and laboratory parameters of 51 MM patients who were newly diagnosed with MM and had complete follow-up data in the Department of Hematology, Jiangning Hospital Affiliated to Nanjing Medical University, Nanjing From November 2011 to October 2012. Results: Of the 51 patients, 29 were male and 22 were female. Followed up to December 2019, 21 died and 30 survived. The univariate analysis showed the prognosis was influenced by factors including age, disease stage, standard treatment, new drugs, maintenance treatment, hypercalcemia, globulin, albumin, hemoglobin. Patients aged Conclusion: Patients with blood calcium ≥ 2.6mmol/L, hemoglobin Key words multiple myeloma; overall survival; non-genetic; prognostic factors Disclosures No relevant conflicts of interest to declare.

Background: Minimal residual disease (MRD) is becoming standard diagnostic care for multiple myeloma. Here we investigate cell-free DNA chromosomal instability as minimal invasive biomarker for minimal residual disease monitoring. Methods: 55 patients were recruited, including 47 newly diagnosed and 8 relapsed multiple myelomas. Plasma samples were collected before treatments, end of 2 cycle and 4 cycle of treatments. Treatment response was assessed by using IMWG criteria. Cell-free DNA was analyzed by illumine HiSeq X10, followed by chromosomal instability analyses by a customized bioinformatics workflow, ultrasensitive chromosomal aneuploidy detector (UCAD), and cfDNA CIN responding to treatment was summarized as cfDNA MRD INDEX. Results: In this cohort study, 53 (96.3%) patients were found with treatment responses after 4 cycle of treatments, including 16 (29.1%) complete, 13 (23.6%) very good partial, 17 (30.9%) partial and 6 (10.9%) marginal responses. The other 2 (3.64%) recurrent MM experiences disease progression. At baseline, plasma cfDNA chromosomal aberrations were found in 14/17(82.3%) multiple myeloma patients. Less patients were found with detectable chromosomal changes after treatments (58.8% after C2, 41.2% after C4, versus 82.3% before treatments, P Furthermore, cfDNA CIN MRD index were found linearly correlated with percentage of malignant plasma cells from bone marrow aspiration as examined by flow cytometry assay (R-square=0.883, P Conclusions: Plasma cfDNA CIN might be used for monitoring multiple myeloma patients treatment responses, especially for predicting minimal residual diseases. Disclosures No relevant conflicts of interest to declare.

Natural killer (NK) cells are the main innate immune cells, and have powerful antitumor activity in vitro, but NK cells based immunotherapy does not often get satisfactory results in clinical practice due to insufficient number and poor activity of NK cells.. Therefore, how to improve expansion efficiency and killing activity of NK cells in vitro is critical. Here we report an in vitro culture system culturing peripheral blood mononuclear cells (PBMC) with irradiated LCL as feeder cells in 10%FBS/RPMI1640 culture medium in the presence of 100U/ml IL-2 and 5ng/ml IL-15 for 14 days could selectively expand NK cells. With this culture system, the purity of NK cells increased from 11.5%± 7.01% to 75.3±10.9% (P< 0.01), and the expansion efficiency was about 625±63 times than the original PBMC. Compared with the fresh NK cells isolated from PBMC, there was no significant change in the expression of inhibitory receptors on the surface of expanded NK cells, while the expression of activating receptors was significantly increased (p Figure Disclosures No relevant conflicts of interest to declare.

ObjectivePRP(platelet rich plasma) promotes wound healing by various growth factors including VEGF, TGF-β, PDGF and EGF, all those growth factors are released by activated platelets. MethodsPRP is applied externally to the necrotizing fasciitis wound after activated with 10% calcium gluconate, then with dressing fixed. PRP is used once a day after the wound is cleaned. The wound area is measured once, wound tissue type and drainage is recorded. Granulation area and percentage are calculated. ResultsThe case one patient underwent negative pressure drainage treatment before the treatment of PRP. The wound area was 49.41cm2, involving the entire skin layer, with little fluid exudation. The skin around the wound was dry and curving. After 6 times of PRP treatment, the wound was reduced to 15.4cm2 without exudation, and the tissue type was fresh granulation, with 100% granulation proportion. The wound was kept clean after discharge, and reduced to 1.25cm2 after 1 month without special treatment. The wound was fully healed after 2 months, and there was no wound or rupture after the follow up visit of 12 months. The case two patient underwent negative pressure drainage treatment on the wound surface. The wound area was 106.25cm2 and the depth was 1.0cm. The whole skin was involved with stealth. After 21 times of PRP treatment, the wound was reduced to 64.35cm2, and the tissue type was carrion + granulation, which accounted for 85.73%. After autologous skin graft, 100% survived, and the wound was followed up without rupture 10 months later. ConclusionPRP is a safe and effective method for the treatment of necrotizing fasciitis and promotes wound healing through the release of various growth factors by α-particles in activated platelets. PRP provides a new and effective treatment method for the treatment of necrotizing fasciitis by creating conditions for skin graft, shortening the length of hospital stay, and even eliminating the need for skin graft treatment for some wounds. Key words: PRP, necrotizing fasciitis, wound Disclosures No relevant conflicts of interest to declare.

Objective: To explore the significance of the ratio of neutrophils to lymphocytes (NLR), monocytes to lymphocytes (MLR), and platelets to lymphocytes (PLR) in the prognosis of patients with newly diagnosed multiple myeloma. Methods: We retrospectively reviewed the data for 60 multiple myeloma patients who were diagnosed in Jiangning Hospital Affiliated to Nanjing Medical University from August 2011 to March 2020. According to NLR、MLR、PLR, the patients were divided into the low NLR group (NLR Key Wordsmultiple myeloma; overall survival; NLR; PLR; MLR Disclosures No relevant conflicts of interest to declare.

Objective:Tumor targeting proteins were modified on the surface of platelets and chemotherapeutic drugs were encapsulated in platelets. Based on the fact that platelets can enter the tumor environment and interact with tumor cells, the functionalized platelets carrier has dual targeting effect to kill tumor cells.Methods:Aspirin was used to inhibit platelets aggregation and deformation in vitro to obtain complete platelets. Exogenous Tf (transferrin) was modified on platelets surface to target specific tumor cells (human multiple myeloma cell line RPMI8226). DOX (doxorubicin) was loaded into platelets. To verify the inhibitory effect of functional platelets vector on cell. To observe the distribution of targeted functional platelets vector and treatment effect on implant tumor in mice.Results:Aspirin can inhibit platelets aggregation and deformation during the preparation process effectively. Stable platelets can be modified by Tf and encapsulate DOX effectively. At the same time, the functional platelet vector can specifically aggregate in the implant tumor site in vivo and achieve effective anti-tumor effect.Conclusion:Platelets aggregation and deformation can be inhibited by aspirin effectively, which makes it a stable natural drug carrier. The antitumor effect can be achieved by the functional platelets. Keywords:platelets, encapsulation, doxorubicin, tumor Disclosures No relevant conflicts of interest to declare.

Background:Mantle cell lymphoma (MCL) is the secondary common B cell lymphoma subtype that comprises 6 to 8% of non-Hodgkin's lymphoma, and is closely related to the poor clinical outcomes. Previous studies have focussed on the mechanisms mediating ibrutinib resistance, the first-in-class oral covalent inhibitor of Bruton's tyrosine kinase (BTK). The aims of the this study is to identify key genes related to the FGFR1 Knockdown in Mantle cell lymphoma cell line (Z-138) that has been proved to play a vital role in MCL progression. Methods:GSE138127 mRNA microarray datasets from Gene Expression Omnibus (GEO) were analysed to obtain differentially expressed genes (DEGs) between vector control and the Knockdown of FGFR1 charactered by ibrutinib resistance. The GO and GSEA were carried out by WEB-based GEne SeT AnaLysis Toolkit (WebGestalt) to do the functional enrichment analysis. The network analysis of protein-protein interactions (PPIs), TF-gene interaction were carried out by Network Analyst 3.0 to identify hub genes. And the main hub gene was probed using Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway analysis. Results:In total, 175 DEGs were obtained, of which 87 and 88 were up- and down-regulated, respectively. Three hub genes (CDK1, CCND1) were identified and associated to cell cycle and DNA replication. FOXC1 were identified as the potential Transcription factors in the biological process. Conclusion:CDK1, CCND1 may affect the cell cycle regulated by FOXC1, and represent the new candidate molecular markers of the occurrence of ibrutinib resistance. Keywords: Mantle cell lymphoma (MCL), ibrutinib resistance, Network Analyst, Microarray, Protein-protein interactions, Molecular markers Disclosures No relevant conflicts of interest to declare.

Platelets are small pieces of cytoplasm that have become detached from the cytoplasm of mature megakaryocytes (MKs) in the bone marrow. Platelets modulate vascular system integrity and serve important role, particularly in hemostasis. With the rapid development of clinical medicine, the demand for platelet transfusion as a life‑saving intervention increases continuously. Stem cell technology appears to be highly promising for transfusion medicine, and the generation of platelets from stem cells would be of great value in the clinical setting. Furthermore, several studies have been undertaken to investigate the potential of producing platelets from stem cells. Initial success has been achieved in terms of the yields and function of platelets generated from stem cells. However, the requirements of clinical practice remain unmet. The aim of the present review was to focus on several sources of stem cells and factors that induce MK differentiation. Updated information on current research into the genetic regulation of megakaryocytopoiesis and platelet generation was summarized. Additionally, advanced strategies of platelet generation were reviewed and the progress made in this field was discussed.

The mitochondria-mediated apoptosis pathway is an effective option for cancer therapy due to the presence of cell-suicide weapons in mitochondria. However, anti-apoptotic proteins that are over-expressed in the mitochondria of many malignant tumors, such as Bcl-2 protein, could allow the cancer cells to evade apoptosis, greatly reducing the efficacy of this type of chemotherapy. Here, we constructed a hierarchical targeted delivery system that can deliver siRNA and chemotherapeutic agents sequentially to tumor cells and mitochondria. In detail, the copolymer TPP-CP-LND (TCPL) was synthesized by the mitochondria-targeting ligand triphenylphosphine (TPP) and therapeutic drug lonidamine (LND) conjugated to the polyethyleneimine in chitosan-graft-PEI (CP), and then complexed with siRNA. Followed, the complexes were coated with poly(acrylic acid)-polyethylene glycol-folic acid (PPF) copolymer to form a hierarchical targeted co-delivery system (TCPL/siRNA/PPF NPs). The TCPL/siRNA/PPF NPs had a neutral surface charge, were stable in plasma and exhibited pH-responsive shell separation. Remarkably, the TCPL/siRNA/PPF NPs simultaneously released siBcl-2 into the cytoplasm and delivered LND to mitochondria in the same cancer cell after FA-directed internalization, and even synergistically activated mitochondria apoptosis pathway. This work demonstrated the potential of RNA-interference and mitochondria-targeted chemotherapeutics to collaboratively stimulate the mitochondria apoptosis pathway for cancer therapy.

Garcinia, a kind of dry resin secreted by Garcinia hanburyi Hook. F. G., is a traditional Chinese medicine with various biological functions such as detoxification, anti-inflammatory, and anthelmintic activities. Recent studies suggest that garcinia has potential anticancer activity. Increasing evidences indicate that the main active monomer gambogic acid isolated from garcinia can inhibit the growth of various cancer cells. Neogambogic acid is an isolated compound with a similar chemical structure as gambogic acid. Preliminary studies show that the neogambogic acid can selectively inhibit the growth of various cancer cells, and has a broader antitumor activity and lower toxicity than gambogic acid. In this review, we summarize the advances made in the investigation of the anti-tumor effect of neogambogic acid in recent years.

Background:T-cell malignancies are characterized by the excessive proliferation of hematopoietic precursor cells of T-cell lineage lymphocytes in bone marrow. T-cell malignancies consist of T-cell lymphoblastic leukemia (T-ALL) and T-cell non-Hodgkin lymphoma (T-NHL). Effective therapeutic options are limited for T-cell malignancies patients. Apoptosis is one of the most important means to cure cancer. Autophagy, a conserved process that sequesters and targets cellular components for lysosomal degradation, also plays highly context-specific roles in mediating cell death. Our research found that LW-218, a new synthetic flavonoid compound, exerted anti-T-cell malignancies cell lines effects by autophagy-mediated apoptosis induction. Methods: In vitrostudy, T-ALL cell lines (Jurkat, Molt4) and T-NHL cell lines (Hut102, Hut78) were used to investigated LW-218-induced apoptosis effects by survival assays (Flow cytometry, Annexin V/PI). A series of experiment were used to assess the autophagy flux triggered by LW-218, by using mCherry-GFP-LC3 construct, Lysotracker Red staining, and determining the expression ratio of LC3-II/LC3-I. Western blot assay, RT-qPCR, transfection with siRNA, and immunofluorescence were used to investigate the underlying mechanism. In vivostudies, the anti-tumor effects of LW-218 were evaluate in T-ALL xenografts which established by subcutaneous injection of Jurkat cells. Results:Mechanistic studies demonstrated that LW-218-induced cell apoptosis in T-cell malignancies cell lines was correlated with activation of caspase 8, caspase 3 and caspases 9, and the substrate PARP1. In addition, LW-218 induced autophagy flux in T-cell malignancies, as increased ratios of LC3B-II/LC3B-I and reductions of P62 expression. The increased numbers of GFP-mCherry puncta and fluorescent intensity also indicated the increase in autolysosomes. The autophagy-mediated cell death must strictly follow the criteria that the inhibition of autophagy, through either genetic or chemical means, prevents cell death. LW-218-induced cell apoptosis and the activation of caspases3/8/9 could be prevented when the autophagy flux was blocked by autophagy inhibitors. mTOR is a master regulator of cellular metabolism and governs the programmed cell death pathways of apoptosis and autophagy. The results showed that LW-218 inhibited the expression of p-mTOR (Ser2448). And the restored activation of mTOR correlated with the inhibition of autophagy flux, which indicated mTOR might be involved in LW-218-induced autophagy. LW-218 also induced cleavage of Bid, the substrate of caspases 8, which indicated that the apoptosis mediated by autophagy might via caspases 8 and Bid activation. Thein vivostudies also verified that LW-218 inhibited the growth of tumors. Conclusions:Our study provides a new insight into the mechanism of LW-218-induced autophagy-mediated apoptosis, suggesting the potency of LW-218 to be a promising agent against T-cell malignancies. Disclosures No relevant conflicts of interest to declare.

Background: Prophylactic transfusion of platelets provides a good protection for the implementation of invasive procedures and the prevention of bleeding events during chemotherapy and hematopoietic stem cell transplantation in patients who are suffering from hematological diseases. The issues that what is the optimal dose of platelet transfusion and how to monitor platelet transfusion efficiency are important due to the short storage time of platelet products, rising clinical demands, and decreasing donors. It is worthy that there are amount of studies have been conducted in the past to explore factors that may affect the clinical efficacy of platelet transfusion and characteristics of patients under these different transfusion effects. While, previous studies failed to exactly address the problems of clinical needs for platelet transfusion. Methods: The aim of our study is to develop a model to evaluate the efficacy of platelet transfusion and the statistic method of machine learning algorithm (ML) was involved. The differences between this algorithm and traditional methods are that the former one can continuously be learning from the data and form a self-training model, therefore ML is more accurate than traditional artificial models and generally independent of the model and parameters themselves. We further take the multi-layer fully connected layer neural network model (MLNN) into our consideration because it simulates the multi-layer interconnection of human nervous systems, which is suitable for processing inaccurate and fuzzy information. In our study, the establishment of a neural network model was used to make a multiple-dimension analysis between factors affecting platelet transfusion efficacy and platelet count added value correction index (aCCI), and explore the correlation among these influencing factors as well. Results: The study utilized the data relative to 1840 platelet transfusions performed in 460 patients with hematological diseases. The participants ranged in age from 16 to 92, and the median age was 59.5. There were 199 females and 261 men. The whole data was divided to 2 parts, 2/3 of them were analyzed as a training set and the others were used for validating. We selected 30 factors (including patient-related factors and transfusion product-related characteristics) that may affect the efficacy of platelet transfusion except the storage time of platelet (all data < 2 days), and established a model for predicting platelet transfusion efficacy based on the volume of platelet transfusion. After the model was established, it was tested for goodness of fit, and the results showed that the LOSS value tended to be stable. Conclusions: The establishment of this model may not only be used for predicting the platelet count after platelet transfusions in patients and the amount of platelets that need to be transfused, but also provide supports for the solution of related problems. Disclosures No relevant conflicts of interest to declare.

Objective :At present, patients with hematopoietic stem cell transplantation (HSCT) are gradually increasing due to hematological diseases. However, there are no further studies on the factors affecting the effect of platelet transfusion during HSCT. This study analyzed the possible factors that affect the platelet transfusion efficiency. Methods: The clinical data of patients with HSCT were collected from the Department of Hematology, Zhongda Hospital Affiliated to Southeast University from March 2015 to March 2018. According to the inclusion and exclusion criteria, a retrospective statistical analysis on the factors affecting the infusion effect was taken in all patients from +1 d to the day that the peripheral blood granulocyte count was more than 0.5×109/L after transplantation. The factors are including gender, age, types of disease, whether initial treatment, platelet inventory days in the effective period, types of transplantation, duration of disease and transfusion times. Furthermore, non-conditional logistic regression analysis was performed on the mentioned factors with statistical significance. Results: A total of 52 patients, 299 platelet transfusions were included in this study. The patients involve 28 men and 24 women, and there were 233 effective infusion and 66 invalid infusion. After chi-square test, the effects of gender, age, disease types, whether initial treatment, transplantation types, duration of disease and transfusion times on platelet transfusion were statistically significant(P Conclusion: There are many factors related to poor platelet transfusion during HSCT, in which the gender, disease types and whether initial treatment are the main influencing factors. Disclosures No relevant conflicts of interest to declare.

Peipei Xu,1 Jingyuan Li,2 Lixin Shi,3 Matthias Selke,3 Baoan Chen,4 Xuemei Wang51Department of Hematology, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, People’s Republic of China; 2Laboratory Animal Center, Institute of Comparative Medicine, Nantong University, Nantong, People’s Republic of China; 3Department of Chemistry and Biochemistry, California State University – Los Angeles, Los Angeles, CA, USA; 4Department of Hematology, Zhongda Hospital, Southeast University, Nanjing, People’s Republic of China; 5State Key Lab of Bioelectronics (Chien-Shiung Wu Laboratory), Southeast University, Nanjing, People’s Republic of ChinaAbstract: We prepared and studied novel fluorescent nanocomposites based on gambogic acid (GA) and cadmium–tellurium (CdTe) quantum dots (CdTe QDs) modified with cysteamine for purpose of cancer cell labeling and combined treatment. The nanocomposites were denoted as GA-CdTe. Characterization results indicated that the CdTe QDs can readily bind onto cell plasma membranes and then be internalized into cancer cells for real-time labeling and tracing of human liver hepatocellular carcinoma cell line (HepG2) cells. GA-CdTe significantly enhanced drug accumulation in HepG2 cells and inhibited cancer cell proliferation. GA-CdTe nanocomposites also improved the drug action of GA molecules in HepG2 cells and induced the G2/M phase arrest of the cancer cell cycle, promoting cell apoptosis. Given the sensitive, pH-triggered release of GA-CdTe, the side effects of GA anticancer agents on normal cells/tissues in the blood circulation markedly decreased. Efficient drug release and accumulation in target tumor cells were also facilitated. Thus, the fluorescent GA-CdTe offered a new strategy for potential multimode cancer therapy and provided new channels for research into naturally-active compounds extracted from traditional Chinese medicinal plants.Keywords: cadmium-tellurium quantum dots, gambogic acid, drug delivery, cell labeling, therapy

Autophagy is a normal physiological process necessary for cellular homeostasis to maintain adequate levels of cellular components. It is essential to stabilize the source of energy during development and nutritional stress and plays the dual role of survival or cell killing in various diseases including cancer. The selectivity of the response to removal of selected organelles may vary according to the each type. Macroautophagy forms a double-membraned autophagosome around the organelle destined for processing. Microautophagy involves direct engulfment of the cellular components by lysosomal invagination. Chaperone mediated autophagy (CMA) is highly selective and is dependent on the chaperone hsc70 for its activity. The effects of all these types are implemented by autophagy related genes. In this review, the markers, activators, inhibitors biological effects and roles of the three classes of autophagy in cancer and obesity are discussed.

Lianghua Fang,1,3 Baoan Chen,2 Shenlin Liu,3 Ruiping Wang,3 Shouyou Hu,3 Guohua Xia,2 Yongli Tian,3 Xiaohui Cai21No 1 Clinical Medical College of Nanjing University of Chinese Medicine, 2Department of Hematology, Zhongda Hospital, Medical School, Southeast University, 3Department of Oncology, Jiangsu Province Hospital of Traditional Chinese Medicine, Nanjing, People's Republic of ChinaBackground: The present study evaluated whether magnetic nanoparticles containing Fe3O4 could enhance the activity of gambogic acid in human colon cancer cells, and explored the potential mechanisms involved.Methods: Cytotoxicity was evaluated by MTT assay. The percentage of cells undergoing apoptosis was analyzed by flow cytometry, and cell morphology was observed under both an optical microscope and a fluorescence microscope. Reverse transcriptase polymerase chain reaction and Western blot assay were performed to determine the transcription of genes and expression of proteins, respectively.Results: Gambogic acid could inhibit proliferation of LOVO cells in a dose-dependent and time-dependent manner and induce apoptosis, which was dramatically enhanced by magnetic nanoparticles containing Fe3O4. The typical morphological features of apoptosis in LOVO cells were observed after treatment comprising gambogic acid with and without magnetic nanoparticles containing Fe3O4. Transcription of cytochrome c, caspase 9, and caspase 3 genes was higher in the group treated with magnetic nanoparticles containing Fe3O4 and gambogic acid than in the groups that received gambogic acid or magnetic nanoparticles containing Fe3O4, but transcription of phosphatidylinositol 3-kinase, Akt, and Bad genes decreased. Notably, expression of cytochrome c, caspase 9, and caspase 3 proteins in the group treated with gambogic acid and magnetic nanoparticles containing Fe3O4 was higher than in the groups receiving magnetic nanoparticles containing Fe3O4 or gambogic acid, while expression of p-PI3K, p-Akt, p-Bad, pro-caspase 9, and pro-caspase 3 degraded.Conclusion: Magnetic nanoparticles containing Fe3O4 can enhance apoptosis induced by gambogic acid which may be closely related to regulation of the PI3K/Akt/Bad pathway in the treatment of human colon cancer.Keywords: Fe3O4, magnetic nanoparticles, gambogic acid, LOVO cells, apoptosis, PI3K, Akt, Bad, pathway

Background: Multiple myeloma (MM) is a plasma cell malignancy characterized by complex cytogenetic and molecular abnormalities including translocations involving the immunoglobin heavy chain locus and mutations involving numerous oncogenic signaling pathways. Fluorescence in situ hybridization (FISH) has emerged as the most useful current cytogenetic assessment and provide a new level of insight into the correlation of myeloma prognosis risk model. However, the identification or sorting of malignant cells is required before FISH probes and only involved expansion of the types of probe and number of detectable targets is reached. Cell-free DNA (cfDNA) offers the potential for minimally invasive genome-wide profiling of tumor alterations without tumor biopsy and may be associated with cancer precision medicine and patient prognosis. Methods: In this retrospective cohort study, we identified 37 patients from 9 relapsed/refractory (RR) and 33 newly diagnosed (ND) patients were analyzed for chromosomal copy number imbalance using the ultrasensitive chromosomal aneuploidy detector (UCAD) platform. Results: Chromosome copy number aberration (CNA) were frequently (82.6%, N=46) detected in MM plasma cell free DNA. Applying UCAD to cfDNA, FISH in CD138 purified bone marrow aspirates, and some matched bone marrow biopsies, we find concordance in copy number alterations (~81%) between liquid and tumor biopsies. Significant copy number changes, including 1q gains, 13q deletion and 17p deletion could be found in 57.89%, 54.05%, and 16.67% in plasma of MM, which is higher percentage than FISH assay (46.81%, 28.26%, and 8.89%), respectively. Besides, chromosome 6p and 6q were determined the higher frequency aberration from UCAD. Moreover, a higher frequency of copy number aberrations and variations was detected in RR patients than ND (100% vs 78.4%, respectively), obviously CNAs heterogeneity displayed in advanced disease. In the inconsistent some samples, UCAD from the plasma and bone marrow showed the similar results, which indicated the FISH is underdetermined and insensitivity in some patients' routine inspection. Conclusion: We conclude that cfDNA analysis as an adjunct to BM biopsy represents a noninvasive and broaden the applicability strategy for comprehensive genomic profiling and therapeutic monitoring of MM. Disclosures No relevant conflicts of interest to declare.

Corresponding author: Baoan Chen, MD, PhD; E-mail: cba8888@hotmail.com Zhuyuan Wang, PhD; E-mail: wangzy@seu.edu.cn Yiping Cui, PhD; E-mail: cyp@seu.edu.cn Keywords: leukemia; multi-drug resistance; P-glycoprotein; surface-enhanced Raman scattering; SERS intensity. Background Acquisition of multidrug resistance (MDR) in the chemotherapy of leukemia could decrease the survival rate of refractory/relapsing patients. One of the best characterized mechanisms of MDR in leukemia is mediated by multidrug resistance protein-1 and its product, P-glycoprotein (P-gp). Thus, accurate detection of P-gp is necessary for MDR diagnosis. In the recent years, surface-enhanced Raman scattering (SERS) has emerged as a new detection technology of biological label for immunoassay with the advantages of ultrasensitive screening ability and extensive adaptability. However, few researches have focused on the application of SERS immunoassay in the diagnostics of leukemia MDR. The aim of our study is to investigate the expressions of P-gp on the cell surface of K562/ADM cells, and in the whole-blood samples of leukemia using a SERS-based immunoassay technique. Methods To simulate the MDR occurrence, we mixed the K562 and K562/ADM cells at different ratios. Besides, we built up the concentration gradient of K562/ADM cells for the quantitative analysis. We also divided 30 blood samples (AML n=14, ALL n=16; female n=12, male n=18; age Results There were positive and stable SERS signals of peak intensity at 1078 cm-1 after suspended with target samples. First, the SERS intensity of K562/ADM was significantly higher than that of K562 (P Conclusion We have proposed a SERS-based immunoassay to evaluate the expression of P-gp, a product of MDR protein of leukemia. Qualitative and quantitative analysis of K562/ADM indicated excellent specificity, high sensitivity and detection limit, as well as great reproducibility of this immunoassay. It was also demonstrated that this immunoassay was with acceptable accuracy and detection reproducibility for clinical whole blood samples, which was of great importance and convenience for practical clinical application. These features have made SERS-based immunoassay a selective and convenient technique for the identification of leukemia MDR diagnosis. Disclosures No relevant conflicts of interest to declare.

Platinum-based chemotherapeutics are the most common regimens for advanced non-small-cell lung cancer (NSCLC) patients. However, it is difficult to identify platinum resistance in clinical treatment. Genetic factors are thought to represent important determinants of drug efficacy. In this study, we investigated whether singlenucleotide polymorphisms (SNPs) in human mutS homolog 2 (hMSH2) and the human mutL homolog 1 (hMLH1) were associated with the tumor response in advanced NSCLC patients received platinum-based chemotherapy in Chinese population. Totally, 96 patients with advanced NSCLC were routinely treated with cisplatin- or carboplatin-based chemotherapy. The three-dimensional (3D), polyacrylamide gel-based DNA microarray method was used to evaluate the genotypes of hMSH2 gIVS12-6T/ C and hMLH1-1151T/A with peripheral lymphocytes. We found that there was a significantly increased chance of treatment response to platinum-based chemotherapy with the hMSH2 gIVS12-6T/C polymorphism. The 3D polyacrylamide gel-based DNA microarray method is accurate, high-throughput, and inexpensive, especially suitable for a large scale of SNP genotyping in population.

Background Aberrant DNA methylation of the CpG sites of the tumor suppressor gene is closely associated with carcinogenesis. Recently, several studies have indicated the aberrant methylation of E-cadherin gene could be a potential marker for leukemic patients. Method We used bisulfite-modified DNA as a template for PCR amplification, resulting in conversion of unmethylated, but not methylated, cytosine into thymine within CpG islands of interest. The amplified product containing a pool of DNA fragments with altered nucleotide sequences was then hybridized with an oligonucleotide-based microarray. Five sets of oligonucleotide probes were designed to detect the methylation patterns of E-cadherin gene CpG islands in leukemia samples. The results were further validated by methylation-specific PCR (MSP). Results We found that all leukemia samples were methylated at different levels within the target sequences. The specific regions (the CpG sites #16–19 and #20–22) were revealed as hotspots for methylation in leukemic patients. These results showed that the microarray assay could successfully detect methylation changes of E-cadherin gene in leukemia quantitatively. Conclusion The oligonucleotide-based microarray can be a quick and reliable tool to map methylation status in CpG islands. This established microarray could be potentially useful for clinical researches and diagnosis.

Background The appearance of antibodies to platelets in the blood is an important cause of immune thrombocytopenia (ITP), and platelet glycoprotein (GP)-specific antibody detection may be helpful to diagnose this condition. Methods Photonic crystal microspheres with different distinct reflection spectra were coated with anti-GPIIb, -GPIIIa, -GPIb and -GPIX monoclonal antibodies (MoAbs) to create a photonic crystal-encoded suspension array (PCSA). Fluorescein isothiocyanate-labelled goat anti-human IgG was added to detect human IgG simultaneously. The detection results were analysed by fluorescence microscopy. Parallel MoAb immobilization of platelet antigen (MAIPA) was used as a reference test. Both methods were used to analyse 63 clinical samples including serum from 32 ITP patients and 31 healthy humans. Results The PCSA showed greater sensitivity than MAIPA in detecting anti-GPIIb (75.0% vs 31.1%) and GPIIIa (84.4% vs 40.6%) antibodies and similar sensitivity as MAIPA in detecting anti-GPIb (37.5% vs 34.4%) and GPIX (50.0% vs 40.8%) antibodies. The MAIPA and PCSA tests had similar specificity. The PCSA detected higher dilutions of serum containing anti-GPIIIa antibody or anti-GPIIb antibody than did MAIPA. The entire testing process was controlled within 3.5 h. Conclusions The PCSA assay described has comparable or better sensitivity and specificity compared to the MAIPA and is more rapid.

Wen Bao,1 Ran Liu,1 Yonglu Wang,1,2 Fei Wang,1 Guohua Xia,1 Haijun Zhang,1 Xueming Li,2 Haixiang Yin,2 Baoan Chen11Department of Hematology and Oncology, Key Medical Disciplines of Jiangsu Province, Zhongda Hospital, Medical School of Southeast University, 2College of Pharmacy, Nanjing University of Technology, Nanjing, People’s Republic of ChinaAbstract: Chemotherapy offers a systemic cancer treatment; however, it is limited in clinical administration due to its serious side effects. In cancer medicine, the use of nanoparticles (NPs) drug delivery system (DDS) can sustainedly release anticancer drug at the specific site and reduce the incidence of toxicity in normal tissues. In the present study, we aimed to evaluate the benefit of a novel chemotherapeutic DDS and its underlying mechanisms. Daunorubicin (DNR) was loaded into poly (lactic-co-glycolic acid) (PLGA)-poly-l-lysine (PLL)-polyethylene glycol (PEG)-transferrin (Tf) NPs to construct DNR-PLGA-PLL-PEG-Tf-NPs (DNR-loaded NPs) as a DDS. After incubating with PLGA-PLL-PEG-Tf-NPs, DNR, and DNR-loaded NPs, the leukemia K562 cells were collected and the intracellular concentration of DNR was detected by flow cytometry, respectively. Furthermore, the effect of drugs on the growth of tumors in K562 xenografts was observed and the relevant toxicity of therapeutic drugs on organs was investigated in vivo. Meanwhile, cell apoptosis in the excised xenografts was measured by transferase-mediated dUTP nick-end labeling assay, and the expression of apoptosis-related proteins, including Bcl-2, Bax, Caspase-9, Caspase-3, and cleaved-PARP, was determined by Western blotting analysis. Results showed that DNR-loaded NPs increased intracellular concentration of DNR in K562 cells in vitro and induced a remarkable improvement in anticancer activity in the xenografts in vivo. The expression of Bcl-2 protein was downregulated and that of Bax, Caspase-9, Caspase-3, and cleaved-PARP proteins were obviously upregulated in the DNR-loaded NPs group than that in other ones. Interestingly, pathological assessment showed no apparent damage to the main organs. In summary, the results obtained from this study showed that the novel NPs DDS could improve the efficacy of DNR in the treatment of leukemia and induce apoptosis via intrinsic pathway. Thus, it can be inferred that the new drug delivery may be a useful clinical tool.Keywords: PLGA-PLL-PEG, daunorubicin, transferrin, K562 cells, apoptosis

We report a novel approach to enhance the efficient accumulation and utilization of anticancer drug daunorubicin on cancer cells through the combination with CdS nanoparticles. Our observations using confocal fluorescence scanning microscopy as well as electrochemical analysis methods demonstrate that CdS nanoparticles can readily bind with daunorubicin on the external membrane of the targeted cells and facilitate the uptake of drug molecules in the human leukemia K562 cells. Besides, our results also indicate that the competitive binding of CdS nanoparticles with accompanying anticancer drug to the membrane of leukemia K562 cells could efficiently prevent the drug release by the drug-sensitive and drug-resistant leukemia cells and thus inhibit the possible multidrug resistance of cancer cells, which could be further utilized to improve the future drug efficiency in respective tumor chemotherapies.

A novel technique was developed, that was high throughput simultaneous screening of multiple resistance protein expression based on a protein array system. The method combined the advantage of the specificity of enzyme-linked immunosorbent assays with the sensitivity and high throughput of microspot. In this system, the multiple resistance protein arrays were created by spotting the captured antibodies onto the glass slide. The arrays were then incubated with cell samples of leukemia patients. The bound proteins were recognized by biotin-conjugated antibodies and detected by CCD. Experiments demonstrated that three multiple resistance proteins, including Pgp, MRP and BCRP which are members of the ATP-binding-cassette (ABC) superfamily of membrane transporters could be simultaneously detected using this new approach. Research work shows the result is coincident with flow cytometry (FCM) (P>0.01). It provided a methodology to develop many high-density protein array systems to detect a variety of proteins. The protein arrays will provide a powerful tool to identify the leukemia cell protein expression and rapidly validate their MDR determination.

Abstract Text: Objective: The mortality rate of refractory/relapsed acute myeloid leukemia (AML) is very high with poor prognostic. Three are still no standard treatments for these patients nowadays. Our research compared the treatment effect between patients with chemotherapy and transplantation. Method: A retrospective research of 45 patients with refractory/relapsed AML was performed in our center. We analyzed the clinical features including gender, age, classification of AML, performance status (PS), complete remission (CR) duration, cytogenetic and molecular abnormities. Survival analyses were made by using the Kaplan Meier method and took the Log - rank test. Results: The mean survival time of the 45 patients with refractory/relapsed AML was (36.25¡À8.40) months and the median follow up was (9¡À2.58) months. The one year and two years overall survival rate was (40.6¡À7.5)% and (23.7¡À7.0)%. Univariate analysis results demonstrated that age¡Ý60 years and failing to undergo HSCT after relapse had poor influence on OS. Conclusion: Performance of HSCT after relapse can improve the prognosis through the single center study. HSCT is still the effective salvage therapy for patients with refractory/relapsed AML. Our research is benefit for individual therapy and can be applied to improve the survival. Disclosures No relevant conflicts of interest to declare.

Multiple myeloma (MM) is a plasma cell malignancy that accounts for approximately 10% of all hematological cancer. Although over the last few decades significant improvement in outcomes has been observed in MM patients, the prognosis of MM remains unfavorable. Existing agents, including the proteasome inhibitor bortezomib and the immunomodulatory agents thalidomide and lenalidomide, have improved outcomes in patients with RRMM greatly. However MM still remains incurable and continuing treatment escalates in complexity while presenting a special therapeutic challenge for patients who these agents have failed. Carfilzomib, a proteasome inhibitor, was approval in 2012 in the United States for RRMM based on efficacy results. Recently, carfilzomib has become a promising therapeutic approach for relapsed and (or) refractory multiple myeloma (RRMM), but no study has summarized the overall effect of carfilzomib in RRMM. To explore the role of carfilzomib, we performed a meta analysis of all known prospective clinical trials to assess the efficacy of carfilzomib in patients with RRMM. Methods and Materials: A systematic review of publications in the PubMed, Embase, the Cochrane Library and ISI Web of knowledge was performed on September 15, 2015 according to the Preferred Reporting Items for Systematic Reviews and Meta Analysis (PRISMA) guidelines. Accounting for some of the inter-study variation, the random-effects model was chosen for the entire study to increase power and precision regardless of heterogeneity. All statistical analyses were conducted by using the STATA software. Meta analyses were carried out to calculate the overall response rate (ORR), complete response rate (CRR) and clinical benefit rate (CBR) of carfilzomib for RRMM. Results: Seven single-arm pilot studies and one randomized controlled trial (RCT) were included. Eight prospective studies enrolled a total of 1,446 patients with 1,000 evaluable patients. The overall quality of the seven single-arm pilot studies was moderate according to Newcastle-Ottawa scale. In the only randomized controlled trial including 792 patients, 396 patients were treated by carfilzomib with lethalidomide and dexamethasome. The quality of this study was adequate according to Cochrane tool for assessment of bias. In patients with RRMM, ORR was 0.44, CRR was 0.13 and CBR was 0.54. High heterogeneity between studies was observed, and funnel plots was symmetrical, negating publication bias. he safety of carfizomib was deemed good and no long-term complications were reported. In the eight prospective studies selected for this analysis, common adverse effect (AE) of the patients varied in different studies, including fatigue, nausea, anemia, thrombocytopenia, neutropenia, diarrhea, etc. Conclusion: In this comprehensive meta analysis, we evaluated the efficacy of carfizomib in the treatment of RRMM. Our meta analysis of the eight studies included, our results demonstrate that carfilzomib is a safe, effective and well tolerated treatment in a large, well-characterized group of patients with RRMM. The lack of severe toxicities observed in patients treated with carfilzomib indicates the potential for full doses of carfilzomib to be used for patients with advanced MM. Disclosures No relevant conflicts of interest to declare.

Information A simple, novel and fast electrochemical method was applied for clinical diagnosis of leukemia. Blood samples of leukemia patients were identified by screen-printed electrodes with potassium ferricyanide (III) probes. Our study successfully developed and raised the possibility of utilizing the electrodes in the identification and diagnosis of leukemia. Background Accurate preliminary detection of leukemia is vital for timely and suitable chemotherapy and clinical outcomes of patients. Electrochemical methods have been widely used in cytobiology, including identification of leukemia cells. In the present study, we creatively applied the use of an electrochemical method to the analysis of human blood samples. The aim of this study was to identify leukemia, breast cancer, respiratory infection and healthy individuals by detecting the electrochemical characteristics of leukocytes in the peripheral blood of patients with a screen-printed electrode. This method might be effectively used in the preliminary screening of leukemia. Methods A total of 99 blood samples obtained from Zhongda hospital in China (May 2016 - July 2016) were divided into four groups: healthy individuals (n = 25), respiratory infection (n=23), leukemia (n = 29) and breast cancer (n = 22). We used probes to distinguish leukocytes of people's blood from different samples immediately due to the different electrochemical behaviors of leukocytes. A screen-printed electrode was used to measure differential pulse voltammetry by a potassium ferricyanide (III) probe combined with a simple bio-sensor system. Then, one-way analysis of variance (ANOVA) method was applied to analyze the scanning curves and peak potential and peak shifts were compared (ΔEp). Results The scanning curves demonstrated the specific electrochemical behaviors of the blank potassium ferricyanide solution and which mixed with different samples in different groups. Significant differences in mean peak potentials were observed over the four groups (P< = 0.001). 105.00±8.02mV, 111.84±9.53mV, 120.90±11.18mV, 132.84±11.53mV for Group healthy, Group respiratory infection, Group leukemia, and Group breast cancer, respectively. Meanwhile, there were 4.67±3.14mV, 10.91±5.81mV,20.42±8.50mV, 33.42±10.05mV in peak shifts for above four groups respectively. Conclusions Using the screen-printed electrode to identify cancer was a novel method with good sensitivity and specificity. It might be effective and had a potential utility in the preliminary screening of leukemia. Disclosures No relevant conflicts of interest to declare.

Background: Allogeneic stem cell transplantation (allo-HSCT) is the only curative option for several hematological malignancies. Allogeneic stem cell transplantation (allo-HSCT) is the only curative option for several hematological malignancies. Patients with GVHD suffer from considerable organ damage, leading to significant morbidity, frequent hospitalizations and high mortality rates. Unlike the acute GVHD which is mediated by cytotoxic T-cell attack on host tissues, pathophysiology of cGVHD is significantly more complex and the mechanisms of dysregulated adaptive and innate immune responses are poorly understood. Many patients require additional salvage therapy for corticosteroid-refractory GVHD and have a dismal prognosis. In recent years, ECP as a therapeutic option has been widely studied in patients with GVHD. In particular, ECP has demonstrated positive effects in patients with steroid-refractory or steroid-intolerant chronic GVHD. The mechanism of action in ECP still remains to be elucidated. To investigate the evidence for clinical efficacy of ECP in the treatment of steroid-refractory cGVHD, we conducted a systematic search and meta-analysis comprising several databases and abstracts presented at recent annual meetings in the field. Methods and Materials: A systematic review of publications indexed in the PubMed, Embase, the Cochrane-controlled trails registry, the Cochrane Library and ISI Web of knowledge were performed on January 10, 2015 according to the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analysis) guidelines. The use of a random-effects model should account for some of the inter-study variation, and all statistical analyses were conducted by using the STATA software. We estimated relative risk (RR) with their 95% confidence interval (95% CI) using the standardized mean difference (SMD). Heterogeneity was evaluated by using I2 values. Meta-analyses were carried out to calculate the overall response rate (ORR) and complete response rate (CRR) of cGVHD and the organ specific responses. Results: Eight complete peer-reviewed papers including seven single-arm pilot studies and one randomized controlled trial (RCT) met the selection criteria and were selected in this meta-analysis. In patients with cGVHD, pooled ORR and CRRwere 0.66 and 0.46 respectively, with organ specific responses of 0.80 for skin, 0.79 for gut, 0.57 for oral mucosa, 0.57 for liver, 0.50 for eye and 0.46 for joint involvement. The safety of ECP is excellent. No long-term complications were reported when compared to other immunosuppressive therapies. Conclusion: Compared with other meta-analyses, our meta-analysis included the most comprehensive literature in the field of ECP treatment in steroid-refractory cGVHD, collected the most recent papers, thus assessing the largest cohort ever of ECP patients (n=176), and employed modern statistical methods like funnel plots to evaluate heterogeneity of studies on the effect of ECP in patients with cGVHD thoroughly in our analysis, which could better help guide clinicians in the ECP application. Our analysis revealed a promising clinical benefit of ECP for patients with steroid-refractory cGVHD. Further prospective trials with larger cohorts are mandatory. Disclosures No relevant conflicts of interest to declare.

Background: Dynamin-2 (DNM2) is a GTPase essential for intracellular vesicle formation and trafficking, cytokinesis and receptor endocytosis. Mutations in DNM2 are common in early T-cell precursor acute lymphoblastic leukemia (ALL). However, DNM2 expression in other types of ALL is not reported. Ikaros, encoded by IKZF1, is a transcriptor factor functioned as a tumor suppress gene, and its dysfunction is associated with poor survival and high relapse rate in ALL. Casein Kinase II (CK2) inhibition could restore Ikaros function in high-risk leukemia and CK2 inhibitor-CX4945 showed the therapeutic efficacy on high-risk leukemia with human-derived xenograft mouse model. It is still undetermined if Ikaros regulates DNM2 expression in the leukemic cells. Methods: The 151 patients' and 30 volunteers' BM samples were collected between June 2008 and June 2014 at the First Affiliated Hospital of Nanjing Medical University. The ALL diagnosis was made according to the morphologic, Immunophenotypic, cytogenetic, and molecular criteria of WHO Diagnosis and Classification of ALL (2008).Cytogenetic and molecular analyses as previously reported. The DNM2 expression was determined by qPCR in the patients. All the patients were divided into high or low DNM2 expression groups (Q4 vs Q1-3) and the cutoff was determined by SPSS 17.0. For quantitative parameters, overall differences between the cohorts were evaluated using a Mann - Whitney U -test. For qualitative parameters, overall group differences were analyzed using a χ2 test. All statistical analyses were performed using the SPSS 17.0 and P Results: We studied DNM2 mRNA level in adults with B- and T-cell ALL, and found DNM2 is more highly expressed compared with normals in both forms of ALL. High DNM2 expression is significantly associated with poor overall survival (OS), high relapse rate, and leukaemia cell proliferation markers particularly in B-ALL. DNM2 expression is significantly higher in the patients with IKZF1 deletion compared to that of without deletion. Ikaros directly binds the DNM2 promoter in Nalm6 (B-ALL) and CEM (T-ALL) leukemic cells. Ikaros suppresses the transcription of DNM2 with luciferase reporter assay. Retroviral transduction of Ikaros results in the down-regulation of DNM2 in the leukemic cells. CK2 inhibitor, TBB increases Ikaros binding to promoter of DNM2 and suppresses DNM2 expression in an Ikaros-dependent manner in both leukemic cell lines and primary cells. TBB induced-increase of H3K9me3 binding on the promoter of DNM2 was also observed in leukemic cell lines and primary cells. Finally, DNM2 inhibitor-MiTMAB significantly suppresses the cell proliferation of Nalm6 and CEM cells with the WST-1 cell proliferation assay and has significantly synergistic effect with Ck2 inhibitor, CX-4945 in the cells. Conclusion: High DNM2 expression is associated with Ikarosdys-regulation, revealing their potential roles on the development of ALL. DNM2 inhibitor MiTMAB inhibits cell proliferation and has synergistic effect with CK2 inhibitor CX4945 in leukemic cells. Disclosures No relevant conflicts of interest to declare.

Some young multiple myeloma (MM) patients, coined macrofocal MM, have been reported to present with multiple lytic bone lesions and less than 10% bone marrow plasma cells (BMPCs). They usually have low tumor burden and favorable outcome. To further understand the clinical and laboratory features of macrofocal MM, we retrospectively analyzed about 1000 symptomatic MM patients of two centers and found that 49 (4.9%) patients fulfilled the criteria of macrofocal MM. Median age was 54 years; 39 (80%) were male. The immunoglobulin subtype included IgG (13), IgA (5), IgD (4), light chain only (4) and non-secretory type (23). International Staging System (ISS) stage 1 in 45 (92%), and no stage 3 patient. Durie-Salmon (DS) Staging System stage 3A in 48 (98%), and 2A in 1. Extramedullary plasmacytomas (EMPs) in 39 (80%) patients. The mean percentage of BMPCs was 2.1%, and 44 (90%) patients had less than 5% BMPCs. No patients had hypercalcemia and renal impairment. FISH cytogenetics, available in 11 patients, showed IgH translocation in 7, 13q del in 2, t(4;14) in 1, 1q21 amp in 6 and p53 del in 2 patients. Serum free light chain (sFLC), available in 14 patients, showed abnormal sFLC in 7 of 14 patients and 4 in 7 non-secretory type patients. Thirty eight (77.6%) patients received new drug-based induction therapies and 14 patients received autologous stem cell transplantation (ASCT) as consolidation therapy. After a median follow-up of 36 months,the estimated median progression-free survival (PFS) was 41 months and the median overall survival time (OS) had not been reached. Forty four macrofocal MM with ISS stage 1 and DS stage 3A were compared to all (93) typical symptomatic MM patients who were diagnosed at the same period with the same stages according to both ISS and DS Staging System. For macrofocal MM, there were significantly more male patients and EMPs, higher hemoglobin levels and lowerb2-microglobulin (b2M) levels than that of typical MM. There were no significant differences in the proportions of patients received novel agents-based chemotherapy and ASCT between two groups, however, the estimated median PFS of macrofocal MM was significant superior to typical MM (41 vs 23 months, respectively; P= 0.001) (Figure 1), and the estimated median OS was also better in macrofocal MM than typical MM (not reached vs 61 months; P= 0.004) (Figure 2). Multivariate analysis showed that only the type of disease was the independent prognostic factor of both PFS and OS. In summary, Macrofocal MM is a particular subgroup of MM with multiple bone lesions and lower tumor burden, higher frequency of EMPs, and better survival than typical symptomatic MM. Figure 1. Figure 1. Figure 2. Figure 2. Disclosures Fan: The National Natural Science Foundation of China: Research Funding. Hou:Novartis: Membership on an entity's Board of Directors or advisory committees.

Object: The study was aimed to analyze the incidence, risk factors and outcome of invasive fungal infection (IFI) in allogeneic hematopoietic cell transplantation (HSCT) recipients, and to provide evidence for the prevention and treatment of IFI. Method: 79 patients undergoing formed HSCT at this institute from Jan 2005 to Sep 2014 with intact datum, were analyzed retrospectively according to the diagnostic criteria of IFI. The data of age, sex, primary disease, the source of stem cell, HLA-identical situation, ABO matching, conditioning regimen, use of ATG, season, aGVHD, neutropenia less than 0.5×109/L, CMV infection, etiology culture, blood routine test, imaging, relapse of disease and survival were recorded. Moreover, the data of patients with IFI including usage of glucocorticoid and the neutrophil count were recorded when IFI occurred and the blood cells count after IFI. Result: 17 patients has been diagnosed with IFI, in which 13 cases were probable and 4 cases were possible. The infection rate was 21.5%. There were 8 female and 9 male. The median age was 26 years (6-50 years).The total non relapse survival (NRS) rate was 55.6%, The NRS rate in non-infection group was 61.2% and that in infection group was 35.2%. The single-factor analysis showed matching(x2=15.061, p=0.010) and aGVHD (x2=6.983 p=0.008) were risk factors. After IFI, 8 patients died, 2 patients got complete response, 5 patients got partial response and 2 patients got stable response. The death rate was 47%, there was significant difference between two groups. The dose of glucocorticoid before IFI within 30 days was risk factor. Multiple-factor analysis further showed Ⅱ-Ⅳ degree of aGVHD (OR=4.810 p=0.012 CI:1.406-16.459 ) and matching (OR=25 p=0.007 CI:2.380-262.653) were risk factors. Moreover, before IFI, use of methylprednisolone more than 360 mg before infection (OR=0.041 P=0.017 CI:0.003-0.56) was risk factor. Conclusion: Patients with poor matching, unrelated stem cell and Ⅱ-Ⅳ degree of aGVHD are more likely to occur IFI and those who were used more glucocorticoid are likely to die. Therefore, avoiding the risk factors might reduce the incidence rate of IFI and respect using dose of immunosupperssor before infection within 30 days may improve the patients' prognosis. Disclosures No relevant conflicts of interest to declare.

Background: MCM7 is one of the subunits of the MCM2-7 complex that are essential for DNA replication licensing and control of cell cycle progression. It also participates in mRNA transcription and DNA damage regulation. It is found that MCM7 was highly expressed in many cancer cells including Leukemia. Thus, lentivirus-mediated siRNA targeting MCM7 was used to suppress its endogenous expression in K562 cells and develop a novel therapeutic strategy for leukemia. Method: Lentivirus-mediated siRNA targeting MCM7 were designed and infected K562 cells. Down-regulation of MCM7 mRNA and protein expression in K562 cells were determined by Real-time PCR and Western blot analysis. The proliferation of the transfected K562 cells was assessed by CCK8 assay. Cell cycle progression and apoptosis were calculated by flow cytometry. The related protein levels of the transfected K562 cells were detected by Western blot. The stably transfected K562 cells were injected into nude mice. The transplanted tumors were measured and weighed, then Western blot and Immunocytochemistry were used to evaluated the expression of the proteins for mechanism. Results: The mRNA and protein expression of MCM7 in the stably transfected K562 cells decreased with the percentage of 92.3±1.88 and 91.18%±0.93 respectively. Compared with control and negative control groups, the proliferative rate of K562 cells was significantly reduced after LV-MCM7-RNAi infection. The cell cycle progression of K562 cells was blocked, with a massive increase in the percentage of cells in the G0/G1 phase and decrease in S phase. The apoptosis rate of the transfected K562 cells was significantly higher. PLK1 and pPLK1 protein were down-regulated in response to MCM7 inhibition; P53 and bax protein were up-regulated simultaneously. In vivo, the tumor size and weight dramatically decreased in the MCM7-RNAi group than in negative control group. PLK1, pPLK1 proteins in transplanted tumors were down- regulated and P53,bax were up-regulated correspondingly. Conclusions: Lentivirus-mediated MCM7 shRNA has effectively down regulated the expression of MCM7 gene on mRNA and protein levels. Moreover, the silence of MCM7 has led to the significant decrease in proliferation, blocking the cell cycle progression of K562 cells in vitro. Simultaneity, the apoptosis of the leukemia cell was enhanced. Both of the functions above cause the inhibition of the tumor growth in nude mice. Overall, these results suggest that the proliferation and cell cycle of K562 cells could be regulated by silencing MCM7 gene which is a promising gene therapeutic method to treat gastric cancer. Disclosures No relevant conflicts of interest to declare.

A new multiplex chemiluminescent immunoassay (CLIA) based on silica colloidal crystal beads (SCCBs) was developed for tumor marker detection. As the code is the characteristic reflection peak originating from the stop-band of colloid crystal, they avoid photobleaching, the potential interference of encoding fluorescence with analyte-detection fluorescence and chemical instability. Meanwhile our SCCBs suspension array improved the luminescence analysis efficiency by using chemiluminescent detection of enzyme labels. By forming a sandwich immunocomplex on SCCBs, the proposed suspension array was used for simultaneous multiplex detection of tumor markers in one test tube. The results showed that the linear range was 0.5-100ng ml(-1) and 1.0-120ng ml(-1) for carcinoembryonic antigen (CEA) and alpha-fetoprotein (AFP) with a detection limit of 0.12ng ml(-1) and 0.16ng ml(-1) at 3sigma. The proposed array showed the storage stability and the accuracy for sample detection were acceptable, and the results were in acceptable agreement with the reference electrochemiluminescence method. This technique provided an automated, simple, sensitive and low-cost approach for multianalyte immunoassay.

DNA repair capacity (DRC) is correlated with sensitivity of cancer cells toward platinum-based chemotherapy. We hypothesize that genetic polymorphisms in DNA repair gene XPA (xeroderma pigmentosum group A) and XPG (xeroderma pigmentosum group G) (ERCC5, excision repair cross-complementation group 5), which result in inter-individual differences in DNA repair efficiency, may predict clinical response to platinum agents in advanced non-small cell lung cancer (NSCLC) patients. In this study, we find that the Aright curved arrow G change of XPA A23G polymorphism significantly increased response to platinum-based chemotherapy. Polymorphism in XPG His46His was associated with a decreased treatment response, but was not statistically significant.

A novel photonic suspension array was developed for multiplex immunoassay. The carries of this array were silica colloidal crystal beads (SCCBs). The codes of these carriers are the characteristic reflection peak originated from their structural periodicity, and therefore they do not suffer from fading, bleaching, quenching, and chemical instability. In addition, because no dyes or materials related with fluorescence are included, the fluorescence background of SCCBs is very low. With a sandwich format, the proposed suspension array was used for simultaneous multiplex detection of tumor markers in one test tube. The results showed that the four tumor markers, α-fetoprotein (AFP), carcinoembryonic antigen (CEA), carcinoma antigen 125 (CA 125) and carcinoma antigen 19-9 (CA 19-9) could be assayed in the ranges of 1.0–500 ng mL −1 , 1.0–500 ng mL −1 , 1.0–500 U mL −1 and 3.0–500 U mL −1 with limits of detection of 0.68 ng mL −1 , 0.95 ng mL −1 , 0.99 U mL −1 and 2.30 U mL −1 at 3 σ , respectively. The proposed array showed acceptable accuracy, detection reproducibility, storage stability and the results obtained were in acceptable agreement with those from parallel single-analyte test of practical clinical sera. This technique provides a new strategy for low cost, automated, and simultaneous multiplex immunoassay.

The enhancement effect of 3-mercaptopropionic acid capped gold nanoparticles (NPs) in drug delivery and as biomarkers of drug-resistant cancer cells has been demonstrated through fluorescence microscopy and electrochemical studies. The results of cell viability experiments and confocal fluorescence microscopy studies illustrate that these functionalized Au NPs could play an important role in efficient drug delivery and biomarking of drug-resistant leukemia K562/ADM cells. This could be explored as a novel strategy to inhibit multidrug resistance in targeted tumor cells and as a sensitive method for the early diagnosis of certain cancers. Our observations also indicate that the interaction between the functionalized Au NPs and biologically active molecules on the surface of leukemia cells may contribute the observed enhancement in cellular drug uptake.

In multiple myeloma (MM), the hypoxic environment is an important factor causing tumor angiogenesis which is strongly correlated to disease progression and unfavorable outcome by activating the key transcription factor, hypoxia-inducible factor-1α (HIF-1α). Gambogic acid (GA) is the major active ingredient of gamboge, which has been shown to possess antitumor effect by in vitro and in vivo study. However, the underlying molecular mechanism that whether GA inhibits tumor angiogenesis remains not well understood. In this study, we investigated the effects of GA on expression of HIF-1α, and its downstream target gene vascular endothelial growth factor (VEGF) in human MM U266 cells. We found that hypoxia induced increase in the level of HIF-1α subunit protein and activated the PI3K/Akt/mTOR pathway. Moreover, the treatment with GA markedly decreased HIF-1α and VEGF expression under hypoxic condition. Mechanistic studies exhibited that GA inhibited the production of HIF-1α by reducing phosphorylation of Akt and mTOR in U266 cells. Furthermore, in vivo study revealed that intravenous injection of GA once every other day for two weeks could suppress tumor volumes by antiangiogenesis activity. Taken together, our results identify GA suppress hypoxia-activated pathways that linked to MM progression, at least partly, by the inhibition of PI3K/Akt/mTOR signaling pathway. Therefore, GA may be a new potent therapeutic agent against human MM cells. Disclosures No relevant conflicts of interest to declare.

To analysis the cytomegalovirus（CMV） infection rate and median time which the CMV-DNA was positive for the first time. Simultaneously, to demonstrete the risk factors associated with CMV infection and CMV disease distribution and mortality after allogeneic hematopoietic stem cell transplantation（allo-HSCT).A retrospective study among 108 patients with allogeneic hematopoietic stem cell transplantation allogeneic hematopoietic stem cell transplantation(allo-HSCT) was conducted during 1st Jan 2004 and 31stMar 2014 in Zhongda hospital.The infection rate of CMV was 52.78%(57/108), the median time of CMV infection was 44d, especially during 30d~100d. The univariate analysis showed that CMV infection rate was associated with the HLA-identical situation between the donor and recipient, whether the use of Anti-human thymus globulin(ATG) conditioning regimen, neutropenic period after transplantation exceeded 10d and graft-versus-host disease (GVHD). Multivariate analysis showed that CMV infection rate was associated with neutropenic period after transplantation which exceeded 10d and graft-versus-host disease (GVHD).The mortality of the patients who had CMV infection was 10/17 and the mortality of CMV interstitial pneumonia was higher. CMV infection was one of the most commonly happened infection after allo-HSCT. To reduce the incidence of CMV disease and mortality, the best choice of Allo-HSCT was HLA-identical donor, ATG should be used during the process, and neutropenic period should be reduced less than 10 days. Moreover, it is necessary to strengthen the treatment of CMV infection actively. Disclosures No relevant conflicts of interest to declare.

The objective of the present study was to investigate the reversible effect of HZ08 and daunorubicin(DNR) combined with dimercaptosuccinic acid modified iron oxide (DMSA-Fe3O4) magnetic nanoparticles (MNPs) in human chronic leukemia cell line K562/A02, and the mechanism potentially involved. The growth inhibition rate of K562/A02 cells was determined by Cell Counting Kit-8 (CCK-8) assay. Cell apoptosis and intracellular concentration of DNR were detected by flow cytometry (FCM). DAPI staining was used to view apoptotic cellular morphology. Subsequently, transcription levels of MDR1 mRNA and expression levels of P-glycoprotein (P-gp) and caspase-3 were determined by real time polymerase chain reaction (real-time PCR) and Western blotting analysis, respectively. group clearly exhibited more morphological changes (severe structural alterations) than other groups. In addition, transcription of MDR1 gene and protein expression of P-gp and caspase-3 of K562/A02 cells were regulated at the most remarkable extent in DNR-HZ08-MNPs group when compared with other groups. These findings suggest that the remarkable effects of the novel DNR-HZ08-MNPs on multidrug resistant K562/A02 leukemia cells would be a promising strategy for overcoming multidrug resistance. Disclosures No relevant conflicts of interest to declare.

Objective: In this study, we investigated the influence of adriamycin and wogonin to the expression of miR-155 in Burrkit lymphoma in vitro. We focused on exploring the efficiency and pathway of wogonin to induce the apoptosis of lymphoma cells through the transcription and translation level of miR-155 compared with ADM and seeking for the evidence of the synergistic effect between wogonin and ADM. Methods: (1) Use drugs to interfere the growth of the Raji cells (Adriamycin (ADM, chemical name:doxorubicin), wogonin, ADM+wogonin), observe the cells through the microscope. (2) The apoptosis of Raji cells was tested by FCM. (3) The transcription of miRNA-155 of Raji cells was tested by real-time PCR. (4) The translation of miRNA-155 of Raji cells was tested by Western blot. Results: (5) After being disposed for 24 hours,the experimental groups developed to apoptosis.The statistic significance existed between groups of experiment and control and groups of each experiment.The effect of inducing apoptosis from strong to weak was group of ADM+wogonin,ADM and wogonin. (6) After being disposed for 24 hours,the transcription level of miRNA-155 of the experimental groups was lower than the control group.Compared with ADM,the transcription level of miRNA-155 of wogonin group seemed to be stronger,while the combination of the two drugs was the weakest. (7) After being disposed for 24 hours,the translation level of p85α of the experimental groups was lower than the control group. Compared with ADM, the translation level of p85α of wogonin seemed to be stronger, while the combination of the two drugs was the weakest. Conclusion: The Chinese materia medica preparation wogonin could induce the apoptosis of lymphoma cells and enhance the effect of ADM through PI3K/AKT pathway. Disclosures No relevant conflicts of interest to declare.

Drug resistance is a major obstacle in cancer chemotherapy, especially in the treatment of leukemia. Hypoxia is an inseparable component of the solid tumor microenvironment as well as the bone marrow (BM) microenvironment. In this study, we investigated the effect of the new nanoparticles polyethylene glycol (PEG)-poly L-lysine (PLL)-poly lactic-co-glycolic acid (PLGA) (PEG-PLL-PLGA) modified by transferrin (Tf) combined with daunorubicin (DNR) on K562 cells under hypoxia. We found the IC50 of DNR and DNR-PEG-PLL-PLGA in K562 cells increased under hypoxia, while the IC50 of DNR-Tf-PEG-PLL-PLGA in K562 cells decreased under hypoxia according to the results of MTT assay. The apoptosis of cells and the intracellular concentration of DNR were detected by flow cytometry (FCM). From the datas, we found that the new targeted nanoparticles can overcome hypoxia induced drug resistance of K562 cells. By targeting the transferrin receptor (TfR) on the surface of K562 cells, the new targeted drug led to an increase in intracellular accumulation of DNR. Transcription and Protein of P-glycoprotein (P-gp) and TfR increased in K562 cells under hypoxia, which probably had a relationship with the decreased degradation of the Hypoxia-Inducible Factor-1α (HIF-1α). We further detected the levels of HIF-1α, Bcl-2, Bax and Caspase-3 of K562 cells in transcription and protein under hypoxia regulated by DNR, DNR-PEG-PLL-PLGA and DNR-Tf-PEG-PLL-PLGA, respectively. DNR-Tf-PEG-PLL-PLGA had the best effect when compared with other groups. Furthermore, intravenous injection of DNR-Tf-PEG-PLL-PLGA expressed the best effect of suppressing proliferation of K562 cells injected to nude mice among the three drugs. Therefore, the new drug DNR-Tf-PEG-PLL-PLGA may be a potential cancer therapy for human leukemia. Disclosures No relevant conflicts of interest to declare.