The CRISPR/Cas9 system is an efficient genome-editing system that has been successfully applied in the field of gene therapy. However, clinical applications of the CRISPR/Cas9 system are limited by the delivery method and safety concerns. Extracellular Vesicles (EVs) can be released from almost every type of cell, and they act as shuttles to convey molecules between cells. Here, we used EVs derived from epithelial cells as a biosafety delivery platform for the CRISPR/Cas9 system and modified the EVs with a chimeric-antigen receptor (CAR) to give them selective tropism to tumors. Compared to normal EVs, CAR-EVs accumulated in cancer tumors rapidly and released the CRISPR/Cas9 system targeting the MYC oncogene efficiently, both in vitro and in vivo. Taken together, the combination of EV and CAR was confirmed to be a novel strategy facilitating the use of natural gene therapy platforms in cancer treatment in this proof-of-concept research.

ObjectivesThe chemopreventive drug α-difluoromethylornithine (DFMO) has been shown to have an antinociceptive effect on mechanical allodynia in inflammatory arthritis by directly inhibiting ornithine decarboxylase (ODC) and decreasing polyamine production in inflammatory sites. However, little is known about the effect of DFMO on the immune system of inflammatory arthritis. Here, we investigated the effect of DFMO in a well-established collagen-induced arthritis (CIA) mouse model and explored its effect on the immune system.MethodsThe effect of DFMO on the frequency of myeloid-derived suppressor cells (MDSCs) in the spleens of CIA mice and their associations with disease severity, tissue inflammation and the levels of proinflammatory T-helper (Th) 17 cells in lymphoid tissues were investigated. The effects of DFMO on disease severity and Th17 cells were compared with those of antibody depletion of MDSCs. The arthritis severity was also evaluated by adoptive transfer of MDSCs derived from DFMO- or dH2O-treated mice.ResultsIn this study, we showed that both MDSCs and Th17 cells were significantly expanded in CIA mice. Treatment by DFMO at the onset of CIA suppressed the development of arthritis and decreased the frequency of MDSCs and Th17 cells. MDSC depletion by anti-Gr-1 mAb achieved a similar result, while combination treatment of both methods did not achieve a significant difference compared to either of the single treatments. In addition, the adoptive transfer of MDSCs derived from dH2O-treated mice with CIA restored the arthritis severity of CIA in mice treated with anti-Gr-1 mAb, while the transfer of MDSCs from DFMO-treated mice did not have such an effect.ConclusionsOur results identified DFMO as a potential therapeutic drug for the treatment of inflammatory arthritis.

BCR-ABL1 tyrosine kinase inhibitors (TKIs) are selective therapies for patients with chronic myeloid leukemia (CML) and induce deep molecular response (DMR). However, similar to 60% of patients relapse after the discontinuation of TKIs. Relapse after discontinuation is likely due the inability of TKIs to eradicate CML stem cells (CML-LSCs). In our previous study, 12 out of 22 patients maintained a stable DMR after TKI withdrawal, and we found that fewer patients who were treated with second-generation TKI relapsed compared with those receiving imatinib. Therefore, we hypothesized that second-generation TKIs and imatinib may have different effects on CML-LSCs, which may affect the clinical outcome after TKI discontinuation. To investigate this, we established a TKI discontinuation model in vitro by treating CML CD34(+) cells with imatinib and dasatinib continuously for 72 h and then removing the TKI for 24 h. Colony-forming cell (CFC) assays, apoptosis assessment, and proteomic analysis were then performed. We found that TKI discontinuation resulted in less proliferation and CFC output in dasatinib-treated cells compared with imatinib. However, the dasatinib-treated group exhibited increased apoptosis. In the proteomics analysis, we identified 160 upregulated and 151 downregulated proteins when the two TKI discontinuation groups were compared. Importantly, proteins involved in NAD(+) nucleosidase activity, mitochondrial ATP synthesis coupled proton transport, and oxidative phosphorylation were significantly expressed, which were mainly involved in metabolic processes. In conclusion, we demonstrate that imatinib and dasatinib have clear differential effects on CML-LSCs through the regulation of mitochondria oxidative phosphorylation, which may provide a new target for eliminating CML-LSCs in the context of TKI discontinuation.

OBJECTIVESThe purpose of this study is to evaluate the relative risk (RR) of respiratory adverse events (AEs) among patients with RA treated with TCZ.METHODSDatabases (PubMed, Embase and Cochrane Library) were searched for randomised controlled trials (RCT) comparing the use of TCZ with placebo (PBO) or active comparator agents in adults with RA published until October 28, 2017. Statistical analyses were conducted to calculate the RR of infectious and non-infectious respiratory AEs and severe AEs (SAEs) using random-effects or fixed-effects models based on the heterogeneity of the included studies.RESULTSEight trials were ultimately included. TCZ was associated with an increased risk of infectious respiratory AEs relative to comparator agents (RR 1.53, 95% confidence interval [95% CI] 1.04-2.25) but was not associated with an increased risk of non-infectious respiratory AEs (RR 1.19, 95% CI 0.86-1.64). A subgroup analysis revealed similar results for non-infectious AEs and SAEs in the comparisons of TCZ with MTX and adalimumab (ADA), whereas increased risks of these AEs but not SAEs were observed compared with the PBO.CONCLUSIONSOur meta-analysis did not reveal an increase in the risk of non-infectious respiratory AEs in adult patients with RA who were treated with TCZ compared with other csDMARDs and bDMARDs in RCTs.

T cell function in cancer patients is usually impaired due to the constitutive activation of immune checkpoint inhibitors. This state is known as 'exhaustion' and is often associated with the inefficient control of tumors or persistent infections. In this work, we investigated the role of leukemia cell-derived microvesicles (MVs) in T cell exhaustion. Following incubation with MVs from various sources, all T cell subtypes exhibited the exhaustion phonotype and impaired cytokine secretion in vitro. Mice models also showed the connection between immune checkpoint inhibitors and MV injection. Sequencing and bioinformatics analyses indicated that a number of transcription factors and microRNAs (miRNAs) were attributable to the dysregulation of pathways and exhaustion in T cells. Further work revealed that functional miR-92a-3p, miR-21-5p, miR-16-5p, miR-126 and miR-182-5p in MVs could be delivered into T cells to induce the exhaustion phenotype. SerpinB2, IL-1β and CXCL5, which are mediators of the NF-κB pathway, were identified as the targets of the miRNAs mentioned above. We demonstrated that leukemia-derived MVs could initiate T cell exhaustion via the progressive temporal delivery of multiple exogenous miRNAs into T cells and the subsequent interaction of these miRNAs with their targets. Therefore, MVs can be expected not only to become new indicators of the T cell status in patients but also to be used as novel targets for personalized patient treatment.

Objective:To determine the value of serum α1-acid glycoprotein (AAG) for estimating the activity of rheumatoid arthritis (RA) in patients treated with biological DMARDs.Methods:A total of 84 RA patients treated with etanercept or tocilizumab for at least 24 weeks were included in this study.According to the clinical disease activity index (CDAI) at 24 weeks,they were then divided into low disease activity (CDAI ≤ 10) and medium or high disease activity (CDAI ＞10) groups.Serum levels of AAG,erythrocyte sedimentation rate (ESR),and C-reactive protein (CRP) were analyzed after treatment.Results:Of the 41 etanercept-treated patients,18 were in low activity group and 23 in medium or high disease ac tivity group.Of the 43 tocilizumab-treated patients,22 patients were in CDAI ≤ 10 group and 21 in CDAI ＞ 10 group.AAG,ESR,hsCRP,CDAI and disease activity score 28 (DAS28)-ESR were all significantly decreased in low disease activity group as compared with those in medium or high disease activity group after treatment (P ＜ 0.001),regardless treatment with etanercept or tocilizumab.For patients treated with etanercept,AAG,ESR,hsCRP,CDAI and DAS28-ESR had no significant change in medium or high disease activity group.For tocilizumab-treated group,CDAI and DAS28-ESR had no significant change in medium or high disease activity group as compared with baseline,however,AAG,ESR and hsCRP were reduced significantly.Conclusion:AAG can be considered as a reliable marker for evaluating the disease activity in RA patients treated with etanercept.However,for patients treated with tocilizumab,AAG may not exactly reflect disease activity.

Patients with PD-L1 overexpression in relapsed or refractory lymphoma benefited more from anti-PD-1 therapy. Moreover, treatment with approved PD-1 inhibitors was well tolerated.

Background: Tumor necrosis factor receptor type 1-associated death domain protein (TRADD) mediates programmed cell death signaling as well as the Fas-induced cell death pathway. The downregulation of TRADD is found to be associated with the occurrence of many cancers. The present study was designed to investigate the association between TRADD and clinico-pathologic features as well as its clinical significance in acute myeloid leukemia (AML). Methods: Real-time polymerase chain reaction was performed in 100 new AML, 23 AML complete remission patients, and 20 normal individuals. All statistical analysis was performed using SPSS software. Results: It was found that the expression of TRADD messenger RNA was lower in new AML patients as compared to healthy individuals and complete remission patients (P = 0.00239). Moreover, TRADD messenger RNA levels were associated with clinical factors such as risk classification (P = 0.0023) and complete remission (P = 0.0147). Kaplan-Meier analysis revealed that the AML patients with high TRADD expression had significantly prolonged overall survival and higher complete remission compared with low TRADD expressing patients. Conclusions: It is concluded that downregulation of TRADD may be an independent potential prognostic biomarker in AML.

Ye, Cong MD, PhD2; Geng, Zhe MD, PhD1; Li, Shouxin MD, PhD2; Yu, Fei MD, PhD2 Author Information

Objective To investigate the value of neutrophil-to-lymphocyte ratio (NLR) for estimating treatment response of tocilizumab(TCZ) therapy in rheumatoid arthritis (RA). Methods Sixty RA patients treated with TCZ for more than 24 weeks were divided into stable group(41 cases) and flare group( 19 cases) according to disease activity after TCZ treatment. NLR, erythrocyte sedimentation rate (ESR), C-reactive protein(CRP) and clinical disease activity index (CDAI) were compared before and after treatment between the two groups and between stable and flare period in flare group. Results Gender,age,disease period, NLR, ESR, CRP and CDAI were not significantly different between the two groups at baseline ( P > 0.05 ). Compared with before TCZ therapy, NLR, ESR, CRP and CDAI all decreased significantly after therapy in the two groups during stable period ( P < 0.05 ). However, there were no significant differences of these indexes between the two groups during stable period ( P > 0.05 ). Compared with stable period in flare group, swollen joint count, tender joint count, CDAI, ESR, neutrophil count and its percentage as well as NLR all increased during flare period, while percentage of lymphocyte decreased(P <0.05 ). CRP, PLT, hemoglobin and percentage of lymphocyte did not change significantly (P >0.05 ). 68.4% (13/19) and 57.9% (11/19) of patients during flare period in flare group had higher ESR and CRP respectively. 21.1% (4/19) of them had lower neutrophil count and 47.4% (9/19) had higher lymphocyte count. 94.7% (18/19) of them had higher NLR,and CDAI increased in all of them. Conclusion Compared with CRP and ESR, NLR can be considered as a less expensive, but more convenient,reliable and objective marker for evaluating disease activity in RA patients treated with TCZ along with CDAI.

Acute leukemia characterized by chromosomal rearrangements requires additional molecular disruptions to develop into full-blown malignancy, yet the cooperative mechanisms remain elusive. Using whole-genome sequencing of a pair of monozygotic twins discordant for MLL (also called KMT2A) gene-rearranged leukemia, we identified a transforming MLL-NRIP3 fusion gene and biallelic mutations in SETD2 (encoding a histone H3K36 methyltransferase). Moreover, loss-of-function point mutations in SETD2 were recurrent (6.2%) in 241 patients with acute leukemia and were associated with multiple major chromosomal aberrations. We observed a global loss of H3K36 trimethylation (H3K36me3) in leukemic blasts with mutations in SETD2. In the presence of a genetic lesion, downregulation of SETD2 contributed to both initiation and progression during leukemia development by promoting the self-renewal potential of leukemia stem cells. Therefore, our study provides compelling evidence for SETD2 as a new tumor suppressor. Disruption of the SETD2-H3K36me3 pathway is a distinct epigenetic mechanism for leukemia development.

A homocysteine-independent role for B-group vitamins on venous thrombosis (VT) development has been reported. However, related research findings remain inconsistent. PUBMED, EMBASE, and COCHRANE databases were searched to collect information on all eligible studies to make a meta-analysis about the relationship between B-group vitamins and VT. Literature search results did not suggest a correlation between thiamin, pantothenic acid, niacin, or riboflavin with VT. Based on their correlations in the literature, folic acid, vitamin B12, B6 were considered in the meta-analysis and systematic review. Significant standardized mean differences were obtained for plasma folic acid (-0.55; 95 % CI, -0.75 to -0.36) and vitamin B12 (-0.34; 95 % CI, -0.55 to -0.13). Reduced levels of folic acid and vitamin B12 may be independent risk factors of VT. Moreover, a qualitative systematic review indicated that low level of vitamin B6 was an independent risk factor of VT. Randomized clinical studies of B-group vitamins supplementation showed varying results on VT prevention. Multivitamin supplementation for VT prevention, regardless of homocysteine level, would be of interest. Further prospective clinical studies are needed to provide additional evidence on the clinical benefits of B-group vitamin supplementation for VT.

This study was purposed to detect the mutation of isocitrate dehydrogenase 1 (IDH-1) gene in patients with acute myeloid leukemia (AML) and to explore its clinical significance. The genomic DNA was extracted from mononuclear cells (MNC) of bone marrow or peripheral blood in 205 adult AML patients, the exon 4 of IDH1 gene was amplified by PCR, then the sequencing and comparison were performed. The results showed that IDH1 mutation was detected in 9 (4.39%) of 205 AML patients. There were 6 cases of R132H mutation, 1 of R132L mutation, 1 of R132G mutation and 1 of R132S mutation. Significantly more IDH1 aberrations were detected in AML-M2 (P = 0.002) than other types. And the 9 patients with IDH1 mutation were characterized by low platelet count which was lower than patients with wild type IDH1 (P = 0.003). IDH1 mutation combined with FLT3/ITD mutation was found in 5 cases, c-kit mutation in 1, NPM1 mutation in 2, and IDH1 mutation with CEBPA or WT1 mutation was not found, which revealed a significant interaction between IDH1 mutation and the FLT3/ITD positive genotype or the CEBPA wild-type. IDH1 mutation were detected in 4 of 71 (5.63%) CN-AML. There was no significant difference of IDH1 mutation incidence between the normal and abnormal karyotypes. It is concluded that the rate of IDH1 mutation was 4.39% in Chinese AML patients. IDH1 mutation is significantly associated with AML-M2, lower platelet counts in peripheral blood, FLT3/ITD mutation and CEBPA wild-type, but not with age, white blood cell count in peripheral blood, karyotype, NPM1, c-kit or WT1 mutation.

T cell acute lymphoblastic leukemia (T-ALL) is an aggressive leukemia. However the poor prognosis and low morbidity restrict further analysis of the disease. Therefore there is an increasing demand to develop animal models for identifying novel therapeutic approaches. In this study, we inoculated the anti-mouse CD122 monoclonal antibody conditioned NOD/SCID mice with the leukemia cells from 9 T-ALL patients and 1 cell line via the tail vein. Four of the 9 patients and the cell line were successfully engrafted. Flow cytometry detected high percentage of human CD45(+) cells in recipient mice. Immunohistochemistry showed infiltration of human CD45(+) cells in different organs. Serial transplantation was also achieved. In vivo drug treatment showed that dexamethasone could extend survival, which was consistent with clinical observation. These results demonstrated that we successfully established 5 xenotransplantation models of T-ALL in anti-mCD122 mAb conditioned NOD/SCID mice, which recapitulated the characteristics of original disease.

The characteristics of core binding factor (CBF) leukemia appear to differ between Chinese and Caucasian patients. In this study, we analyzed the biological and clinical characteristics of 76 Chinese CBF leukemia patients out of 425 newly diagnosed acute myeloid leukemia (AML) patients. The frequency of CBF AML was 17.9%. Patients harboring t(8;21) were predominant in CBF AML. The incidence of c-kit mutation in CBF AML was 28.9%. The N822K mutation appeared to be more prevalent in Chinese CBF AML patients. Multivariate analysis showed that c-kit mutation and high white blood cell count could negatively impact overall survival (OS) (HR=2.74 and 6.24, P=0.007 and 0.022, respectively) but did not affect relapse-free survival (RFS). Kaplan-Meier analysis showed a significant difference in both OS and RFS between wild-type and mutated c-kit patients. Although we had included recently reported prognostic indicators in our analysis, our results demonstrated that only c-kit mutation and high white blood cell count had prognostic impact on Chinese CBF AML patients.

Abstract Abstract 781 While serving as an initiating event in the disease etiology, chromosomal translocation alone may not be sufficient to drive a particular phenotype of leukemia. Acute leukemia characterized by rearrangements of the histone methyltransferase gene MLL (mixed lineage leukemia) requires additional mutations to develop the full blown malignancy, yet the molecular basis of cooperating events remains under-studied. We reasoned that monozygotic (MZ) twin pairs discordant for human leukemia are well-matched for both inherited genetic background and tissue-specific events, and thereby somatic mutations that arise in the disease twin may play a more prominent role in leukemogenesis. Using whole genome sequencing in a pair of 3-year old monozygotic twins discordant for MLL leukemia, we identified a MLL-NRIP3 fusion gene and mutations in histone H3 lysine 36 methyltransferase SETD2 in the leukemia twin. Retrovirus-mediated ectopic expression of MLL-NRIP3 in mouse hematopoietic cells was able to induce the same type of myeloid leukemia as the patient's in a transplant mouse model. A relative delay in disease onset suggested the occurrence of cooperating events in addition to the initial hit of MLL-NRIP3, in the development of induced leukemia. SETD2 mutations were recurrent (5.4%) in 241 acute leukemia patients, particularly in those with MLL-rearranged myeloid leukemia (22.2%). The identified SETD2 mutations are loss-of-function in nature, characterized by biallelic and truncating mutations, and accompanied by a global loss of trimethylation of H3K36 (H3K36me3) in the patient leukemic blasts. These data suggest that SETD2 acts as tumor suppressor gene in leukemia development. Functionally, transfection of SETD2 shRNA in MLL-AF9 knockin bone marrow cells resulted in decreased levels of SETD2 expression and H3K36 trimethylation. Notably, SETD2 knockdown significantly accelerated the development of MLL-AF9 leukemia in the mouse bone marrow transplantation experiment. Moreover, SETD2 knockdown yielded a significantly higher number of total colonies through the second and third rounds of serial replating of colony-forming cell (CFC) assay, suggesting that loss of SETD2 increased the self-renewal and proliferation potential of MLL leukemia-initiating cells. Finally, we showed that SETD2 deficiency was able to activate gene expression in MAPK, Jak-STAT and mTOR signaling, and dysregulates multiple metabolic and DNA repair pathways that are known to directly contribute to leukemogenesis. This comprehensive study provides compelling evidence for SETD2 as a novel tumor suppressor for leukemia, and suggests that the disruption of distinct histone modifying enzymes, MLL and SETD2, synergistically promotes the development of human leukemia. In addition, our study illustrates that whole-genome sequencing of phenotypically discordant monozygotic twins provides an effective approach, in combination with mutational analysis in patients and functional assays using experimental models, to uncover disease-causal genes. Disclosures: No relevant conflicts of interest to declare.

Objective Toinvestigatetheimmunophenotypiccharacteristicsofacutemyeloidleukemia(AML) patients with NPM1 mutation. Methods The immunophenotype of 237 newly diagnosed AML patients were detected by flow cytometry. Real-time quantitative PCR was employed to detect the NPM1 mutation. The immunophenotype was then compared between the NPM1 mutated and wild type patients. Results The incidence of NPM1 mutation was 19.0 ％ (45/237) in all AML patients.The NPM1 mutated patients had lower expression of CD34,CD117,HLA-DR,CD15 and CD19 than the wild type patients(all P＜0.05).For AML patients with normal karyotype,the incidence of NPM1 mutation was 37.7 ％ (40/106),and the NPM1 mutated patients had lower expression of CD34,HLA-DR,CD15 and CD7 than the wild type patients(all P＜0.05).The NPM1 mutated patients with normal karyotype had lower expression of CD34 HLA-DR and CD7 in M1 subtype(all P ＜ 0.05); lower expression of HLA-DR and higher expression of CD9 in M2 subtype (all P ＜ 0.05) ; and lower expression of CD117 in M5 subtype compared with wild type patients (P ＜0.05). Conclusion The immunophenotypic characteristics of AML patients are changed by NPM1 mutation. The changes of immunophenotype varied in different FAB subtypes.

Nowadays the role of genetic findings in determining the diagnosis, therapy and prognosis of acute myeloid leukemia (AML) has become more valuable. To improve and validate the detection of clonal chromosomal aberrations in leukemia, we designed a combined application of karyotyping with multiplex reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence in situ hybridization (FISH), and addressed the expression and distribution of fusion genes among the subtypes of Chinese adult patients with de novo AML. Multiplex RT-PCR assays were performed on 477 samples from newly diagnosed AML patients, and cytogenetic data were obtained from 373 of them by R or G banding techniques and those in some cases were confirmed by FISH. The PCR products in some suspected cases were tested by two-directional sequencing. The results showed that except unqualified samples, fusion genes were detected by multiplex RT-PCR in 211 of 474 patients (44.51%), including AML1-ETO, CBFβ-MYH11, PML-RARα, PLZF-RARα, NPM-RARα, MLL rearrangements, BCR-ABL, DEK-CAN, SET-CAN, TEL-PDGFR, TLS-ERG, AML1-MDS1 (EVI-1). In 373 patients, who took both multiplex RT-PCR and karyotype analysis, the detection rate of chromosomal aberrations by using multiplex RT-PCR and karyotyping was 160/373 (42.89%) and 179/373 (47.98%) respectively, and the combination could optimize the detection rate of clonal genetic abnormalities to 216/373 (57.90%). The PCR results from 11 cases "normal" in karyotyping but abnormal in RT-PCR for MLL rearrangements were confirmed by two-directional sequencing. It is concluded that karyotype studies remain the cornerstone for genetic testing; conventional cytogenetics and molecular-based methods are complementary tests for the detection of clonal genetic aberrations in AML, especially for the cryptic or submicroscopic aberrations. Once a genetic marker has been identified by combined analysis, it could be used to monitor residual disease during/after chemotherapy, by quantitative RT-PCR and/or FISH.

OBJECTIVE:To establish xenotransplated mouse model by non-obese diabetic/severe combined immunodeficiency (NOD/SCID) mice with primary myeloma cells.METHODS:The model of xenograft was established in NOD/SCID mice by tail vein injection of mononuclear cells from two end stage multiple myeloma patients, three mice were inoculated for each patient. Mice were monitored weekly for body weight. Two weeks later, the human CD45(+) cells from peripheral blood of mice were evaluated by flow cytometry (FCM). The experiment endpoint was body weight loss up to 20% or had pale, vertical hair and listlessness, then spleen and liver were studied by histologic analysis, the human CD45(+)CD38(+) cells from spleen, lymph node, peripheral blood and bone marrow were evaluated by FCM.RESULTS:Body weight of mice in group patient 1 and group patient 2 decreased seven and five weeks after inoculation respectively; the human CD45(+)CD38(+) cells appeared in the peripheral blood (26 ± 4) and (16 ± 4) days after inoculation in group patient 1 and group patient 2 respectively, and increased by time, reaching (16.2 ± 3.0)% and (31.3 ± 3.5)%, respectively at the endpoint; the spleen, liver and lymph node of both groups enlarged, the typical malignant plasma cells were observed in them. The human CD45(+)CD38(+) cells were detected in spleen, lymph node and bone marrow by FCM.CONCLUSION:Our study successfully established a NOD/SCID mouse model xenotransplated with human primary myeloma cells.

OBJECTIVE: To investigate the diagnostic value of FICTION (Fluorescence Immunophenotyping and Interphase Cytogenetics as a Tool for the Investigation of Neoplasms) technique, combining immunofluorescence and fluorescence in situ hybridization (FISH), to detect genetic aberrations in multiple myeloma (MM). METHODS: Bone marrow samples were collected from 18 MM and 2 plasma cell leukemia (PCL) patients. Probes targeting IgH and MMSET were prepared using a Nick Translation Kit from Bacterial artificial chromosome (BAC) clones. The immunophenotyping was achieved via the CD138 tyramide signal amplification (TSA)-mediated immunofluorescence, followed by FISH with the prepared probes \[t(4;14), t(11;14), t(14;16)\] and the commercial deletion probes (13q and p53) to detect common genetic aberrations in MM. RESULTS: All the 20 samples were assayed with the probes mentioned above, and revealed 4 cases with t(4;14), 6 with t(11;14), 1 with t(14;16), 3 with p53 deletion; and 8 with 13q deletion. The remaining 4 cases had none of the 5 aberrations. CONCLUSION: FICTION technique facilitates the detection of genetic abnormalities of MM in situ; enhances both efficiency and sensitivity of positive detection, thus, could be used as the screening test of molecular diagnosis of MM to guide coming-up risk-adapted therapy and evaluate prognosis.