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Organoid culture is a promising biomedical technology that requires specialized growth factors. Recently, a recombinant L-WRN cell line has been extensively used to generate conditioned medium (L-CM) for organoid culture. Nevertheless, methods for evaluating the stability of the L-WRN cells have been limited. In this study, a novel proteomics-based approach was developed to analyze the secretome of the cells. Serum-free L-CM was lyophilized, precipitated by trichloroacetic acid, and desalted prior to analysis by liquid chromatography–tandem mass spectrometry. Data-dependent acquisition (DDA) was conducted for the untargeted secretome profiling of the cells, and parallel reaction monitoring (PRM) was applied for the targeted quantification of the Wnt3A, R-spondin3, and noggin proteins (WRNs). This study also compared the performance of two types of PRM methods, namely MS1-independent PRM and MS1-dependent PRM, that can be executed on an Orbitrap instrument. The results showed that the growth of mouse intestinal organoids was closely related to the use of L-CM. The composition of L-CM could be markedly affected by the medium collection scheme. A total of 1725, 2302, and 2681 proteins were identified from the L-CM collected on day 5, day 9, and day 13, respectively. The MS1-independent PRM outperformed the MS1-dependent PRM and effectively quantified the WRNs with high repeatability and specificity. In conclusion, by integrating untargeted and targeted proteomics, this study develops a mass spectrometry–based method for the secretome analysis and quality control of the L-WRN cells. The methodology and findings of the present work will benefit future studies on organoids and secretomes. [ABSTRACT FROM AUTHOR]

The accuracy level of short-term load forecasting (STLF) affects the power department's arrangements for unit start-up, shutdown, overhaul, and load dispatching. However, the existing algorithms do not fully consider load volatility and difficulty in setting the algorithm parameters. In this regard, this paper proposes a hybrid model which combines complete ensemble empirical mode decomposition with adaptive noise (CEEMDAN), gated recurrent unit (GRU) with improved gray wolf optimizer (IGWO), namely CEEMDAN–IGWO–GRU (CIG) hybrid algorithm. Firstly, the details and trend information of the load signal are separated by the CEEMDAN algorithm to suppress the interference of load fluctuation. Then, the GRU network optimized by IGWO parameters is used to predict each component, separately. Finally, the complete load forecasting results are obtained by reconstructing the forecasting result of each component. The power load data of a certain area under study is used to verify the CIG model, and the experimental results are compared with other existing algorithms. The experimental results show that the load forecasting results of the CIG model get the high-precision evaluation of 0.6997%, 52.4685, and 38.1891 MW in MAPE, RMSE, and MAE, respectively. Therefore, the parameter optimization ability of the IGWO algorithm can effectively improve the prediction accuracy, and the CIG method can availably suppress the impact of load fluctuation on prediction and has a powerful nonlinear fitting ability. In conclusion, CIG has great potential in establishing a power load forecasting model. [ABSTRACT FROM AUTHOR]

In this work, a hybrid method combining interparticle-potential multiphase LBM, finite difference method and characteristic-line wetting scheme, is developed for pore-scale simulation of superheated vapor displacing liquid, driven by gravity, in a porous geometry. The influence of injected vapor superheat, surface wettability and gravity on two-phase displacement and heat transfer processes is investigated. Results show that in thermal displacement, the vapor-liquid interface is unstable and trapped liquid blobs gradually evaporate into vapor. At low vapor superheat degrees, the displacement efficiency, as compared to isothermal displacement, is significantly improved, but it reduces as the vapor superheat degree rises due to the shift of displacement patterns. At all contact angles considered, the displacement always exhibits viscous fingering, although vapor flowpaths become wider with increasing surface wettability. The final vapor saturation is insensitive to surface wettability, but reducing contact angle leads to a less even temperature distribution inside vapor and a decrease in final average vapor temperature. In addition, increasing gravity is found to reduce vapor front temperature, but it almost has no effect on final average vapor temperature because the final average vapor temperature does not only depend on specific interfacial length but also on fingering number and inlet flow rate. • A hybrid method is developed for superheated vapor flooding in porous media. • The effect of vapor superheat degree, surface wettability and gravity is studied. • The displacement efficiency first rises and then falls with vapor superheat degree. • Decreasing contact angle leads to less even temperature distribution inside vapor. • The injection of superheated vapor is found to enhance interface instability. [ABSTRACT FROM AUTHOR]

To investigate thermolysis kinetics and identify degradation compounds, alliin solutions were heated at 60, 80, and 89 °C. The degradation compounds of alliin were identified by high performance liquid chromatography–mass spectrometry (HPLC–MS), tandem mass spectrometry (MS/MS) and ultra-pressure liquid chromatography–high resolution mass spectrometry (UPLC–HRMS). The results showed that the thermal degradation kinetic of alliin could be described by a first-order reaction and k = 4.38 × 10 17 exp (−142494/RT), where k is the reaction rate constant, min −1 ; R is gas constant; T is the absolute temperature, K. Degraded compounds, including S-allyl- l -cysteine and ethers, such as allyl alanine disulfide, allyl alanine trisulfide, allyl alanine tetrasulfide, dialanine disulfide (cysteine), dialanine trisulfide and dialanine tetrasulfide, were identified by HPLC–MS, MS/MS and UPLC–HRMS. Allyl alanine tetrasulfide was identified for the first time in alliin. The results show that alliin is unstable and significant numbers of organosulfur compounds are generated under high temperature treatment. [ABSTRACT FROM AUTHOR]

• Three pehtze stages in sufu production were characterized by high enzyme activities. • Four aging stages in sufu production were characterized by the presence of diverse targeted volatile compounds. • Protease and esterase were strongly correlated with most of the targeted volatile compounds. • Esterase contributed to the formation of three high molecular weight esters. Dynamic changes and correlations between physicochemical properties, 14 targeted volatile compounds (TVCs) and six groups of enzyme activities during eight production stages of naturally fermented plain sufu were explored. Multiple factor analysis was used to discriminate between and group the samples into three clusters: cluster I comprised tofu and pehtze; cluster II comprised dried pehtze and salted pehtze; cluster III involved the aging stages. Clusters I and II were characterised by higher enzyme activities, while cluster III was characterised by the presence of diverse TVCs. Protease and esterase were strongly correlated with most of the TVCs. Esterase, in particular, contributed to the formation of three high molecular weight esters, namely, ethyl dodecanoate, ethyl (Z)-9-octadecenoate and ethyl (Z , Z)-9,12-octadecadienoate. The enzymes found contributed to the texture and flavour of naturally fermented sufu and will provide a good guide and control for using the enzymes directly to ripen sufu products. [ABSTRACT FROM AUTHOR]

Abstract: The frequency of extramedullary infiltration (EMI) in acute myeloblastic leukemia (AML) is reported up to 40% and most prevalent in myelo-monoblastic and monoblastic subtypes of AML (M4 and M5 according to FAB classification). The majority of patients with EMI suffered poor prognosis. To explore mechanism underlying EMI, we analyzed SHI-1 cells, a highly invasive human acute monocytic leukemia cell line, and found their strong expression of matrix metalloproteinase 2 (MMP-2), membrane type 1 MMP (MT1-MMP) and tissue inhibitor of metalloproteinase 2 (TIMP-2). SHI-1 cells showed higher invasive ability to traverse reconstituted basement membranes (Matrigel) and stronger activation of proMMP-2 than other leukemia cell line such as NB4, K562, U937 and THP-1 cells. When co-cultured with bone marrow stromal cells (BMSCs), the invasive capacity and proMMP-2 activation of SHI-1 cells enhanced remarkably. Furthermore, the inhibition of MMP-2, MT1-MMP, or TIMP-2 by small interfering RNA (siRNA) substantially impaired SHI-1 cells invasion and decreased proMMP-2 activation. In the contrast, up-regulated expression of TIMP-2 for 2–3 folds level increased cell invasion and proMMP-2 activation. These results demonstrated that constitutively high expression of MMP-2, MT1-MMP and TIMP-2 in SHI-1 cells facilitated cell invasion by promoting proMMP-2 activation. Moreover, up-regulation of TIMP-2 exhibited not a repressive but an activating effect on SHI-1 cells invasion. Our study indicated that increasing TIMP-2 in AML patients with EMI may potentially cause adverse effects, particularly in patients containing high levels of MMP-2 and MT1-MMP. [Copyright &y& Elsevier]

Abstract: Besides cytopenia, dysplasia is crucial characteristic of MDS-RA. To summarize the morphological features that contribute to the diagnosis of MDS-RA, 48 RA patients with abnormal karyotype were analyzed for the features of morphological and cytogenetical abnormalities and the relationships between them. 46 MDS-RA patients with normal karyotype and 207 patients with non-MDS anemia were enrolled into control groups. More conspicuous and diverse dysplasia can be found in abnormal karyotype MDS-RA than those in control groups (P <0.05). Apparent dysplasia in granulocyte and megakaryocytoid lineages may provide valuable evidence for the diagnosis. Dysplasia occurred more frequently in patients with severe chromosome abnormalities. [Copyright &y& Elsevier]

Abstract: We have studied forty Chinese adult ALL patients at newly diagnosis, using standard primers and protocols of BIOMED-2 multiplex PCR, to determine the feasibility of Ig and TCR gene rearrangements as diagnostic and patient-specific MRD-RQ-PCR targets for molecular monitoring. Clonal IGH, IGK, IGL, TCRB, TCRG and TCRD rearrangements were found in 86%, 22%, 9%, 19%, 77%, 55% of adult patients with B-lineage ALL, respectively. While in T-ALL, clonal IGH, TCRB, TCRG and TCRD rearrangements were detected in 6%, 83%, 78%, 33% of patients. Several specialties in the pattern of Ig/TCR gene rearrangements in Chinese adult ALL patients in comparison with those reported for children and adult patients in other countries have been noted. These results are useful for further MRD-RQ-PCR detection and quantification for all patients. [Copyright &y& Elsevier]

Abstract: CCAAT/enhancer binding proteins (C/EBPs) are a family of transcription factors that have been implicated in diverse cellular functions such as cellular differentiation and proliferation, and inflammatory processes. C/EBPζ, also known as GADD153, CHOP10, and DDIT3 has been found associated with the development of myxoid liposarcoma and the progression of melanoma. To investigate the correlation of C/EBPζ transcript levels with the development of leukemia, samples from 187 patients with myelodysplastic syndrome (MDS), acute myeloid leukemia (AML), and chronic myeloid leukemia (CML) were examined for C/EBPζ mRNA using real-time quantitative PCR (RQ-PCR). RQ-PCR analysis demonstrated the median levels of C/EBPζ were significantly decreased in MDS, AML, and CML patients compared with normal controls (1.40, 0.96, 2.60 versus 14.69, P <0.0001). Significant differences were also observed between patients with CML and with AML or MDS. These results suggest that the insufficient dosage of C/EBPζ might be involved in the development of leukemia. [Copyright &y& Elsevier]

We obtained a full-length cDNA fragment encoding human O6-methylguanine-DNA-methyltransferase (MGMT) from the liver tissue of a patient with cholelithiasis by RT-PCR and confirmed by DNA sequencing. The polycistronic retrovirus vector G1Na-MGMT-Neor-IRES-MDR1 was constructed and verified by restriction endonuclease analysis and DNA sequencing. The vector was transfected into packaging cells GP+E86 and PA317 by the LipofectAMINE method. Cord blood CD34+ cells were transfected with the supernatant of retrovirus containing human MGMT and MDR1cDNA under stimulation of hematopoietic growth factors. PCR, RT-PCR, Southern Blot, Western Blot, FACS and MTT analyses showed that dual drug resistance genes have been integrated into the genomic DNA of cord blood CD34+ cells and expressed efficiently. The transgene cord blood CD34+ cells conferred 5.8–6.3-fold stronger resistance to P-glycoprotein effluxed drugs and 5-fold to BCNU than untransduced cells. The polycistronic retrovirus vector mediated transfer of two different types of drug resistance genes into human cord blood CD34+ cells and co-expression provided an experimental foundation for improving combination chemotherapy tolerance in clinical practice. [Copyright &y& Elsevier]

Cultured keratinocytes are desirable models for biological and medical studies. However, primary keratinocytes are difficult to maintain, and there has been little research on lingual keratinocyte culture. Here, we investigated the effect of Y-27632, a Rho kinase (ROCK) inhibitor, on the immortalization and characterization of cultured rat lingual keratinocyte (RLKs). Three Y-27632–supplemented media were screened for the cultivation of RLKs isolated from Sprague–Dawley rats. Phalloidin staining and TUNEL assay were applied to visualize cytoskeleton dynamics and cell apoptosis following Y-27632 removal. Label-free proteomics, RT-PCR, calcium imaging, and cytogenetic studies were conducted to characterize the cultured cells. Results showed that RLKs could be conditionally immortalized in a high-calcium medium in the absence of feeder cells, although they did not exhibit normal karyotypes. The removal of Y-27632 from the culture medium led to reversible cytoskeletal reorganization and nuclear enlargement without triggering apoptosis, and a total of 239 differentially expressed proteins were identified by proteomic analysis. Notably, RLKs derived from the non-taste epithelium expressed some molecular markers characteristic of taste bud cells, yet calcium imaging revealed that they rarely responded to tastants. Collectively, we established a high-calcium and feeder-free culture method for the long-term maintenance of RLKs. Our results shed some new light on the immortalization and differentiation of lingual keratinocytes. [ABSTRACT FROM AUTHOR]

In view of the current multi-source information scenario, this paper proposes a decision-making method for electric vehicle charging stations (EVCSs) based on prospect theory, which considers payment cost, time cost, and route factors, and is used for electric vehicle (EV) owners to make decisions when the vehicle's electricity is low. Combined with the multi-source information architecture composed of an information layer, algorithm layer, and model layer, the load of EVCSs in the region is forecast. In this paper, the Monte Carlo method is used to test the IEEE-30 model and the traffic network based on it, and the spatial and temporal distribution of charging load in the region is obtained, which verifies the effectiveness of the proposed method. The results show that EVCS load forecasting based on the prospect theory under the influence of multi-source information will have an impact on the space–time distribution of the EVCS load, which is more consistent with the decisions of EV owners in reality. [ABSTRACT FROM AUTHOR]

To reduce the short‐term load forecasting (STLF) error of off‐line forecasting model, a VMD‐IWOA‐LSTM (VIL) method for STLF is proposed. Firstly, variational mode decomposition (VMD) is used to decompose the historical power load signals. Then, the decomposed signals are reconstructed according to the similarity of Pearson correlation coefficient (PCC), and meteorological data are chosen for each reconstructed component based on the set PCC threshold. The long short‐term memory (LSTM) models are used to predict the corresponding components, and improved whale optimization algorithm (IWOA) is used to optimize the parameters in LSTM. Finally, the forecast results of each component are added together to get the final forecast result. The experimental results of power load data in a certain area show that the proposed method has the advantages of strong anti‐interference performance and high prediction accuracy compared with other methods, and has strong practicability. © 2022 Institute of Electrical Engineers of Japan. Published by Wiley Periodicals LLC. [ABSTRACT FROM AUTHOR]

Background: Fasting is known to influence the immune functions of leukocytes primarily by regulating their mobilization and redistribution between the bone marrow and the peripheral tissues or circulation, in particular via relocalization of leukocytes back in the bone marrow. However, how the immune system responds to the increased risk of invasion by infectious pathogens with fewer leukocytes in the peripheral blood during fasting intervention remains an open question. Results: We used proteomic, biochemical and flow cytometric tools to evaluate the impact of short-term intensive fasting (STIF), known as beego, on red blood cells by profiling the cells from the STIF subjects before and after 6 days of fasting and 6 days of gradual refeeding. We found that STIF, by triggering the activation of the complement system via the complement receptor on the membrane of red blood cells, boosts fairly sustainable function of red blood cells in immune responses in close relation to various pathogens, including viruses, bacteria and parasites, particularly with the pronounced capacity to defend against SARS-CoV-2, without compromising their oxygen delivery capacity and viability. Conclusion: STIF fosters the immune function of red blood cells and therefore, it may be considered as a nonmedical intervention option for the stronger capacity of red blood cells to combat infectious diseases. Keypoints: STIF fosters the immune response of red blood cells to various pathogens, in particular SARS-CoV-2, via activation of the complement receptor on their membranes. STIF does not compromise the oxygen transport capacity or viability of red blood cells. [ABSTRACT FROM AUTHOR]

Background: Fasting is known to influence the immune functions of leukocytes primarily by regulating their mobilization and redistribution between the bone marrow and the peripheral tissues or circulation, in particular via relocalization of leukocytes back in the bone marrow. However, how the immune system responds to the increased risk of invasion by infectious pathogens with fewer leukocytes in the peripheral blood during fasting intervention remains an open question. Results: We used proteomic, biochemical and flow cytometric tools to evaluate the impact of short-term intensive fasting (STIF), known as beego, on red blood cells by profiling the cells from the STIF subjects before and after 6 days of fasting and 6 days of gradual refeeding. We found that STIF, by triggering the activation of the complement system via the complement receptor on the membrane of red blood cells, boosts fairly sustainable function of red blood cells in immune responses in close relation to various pathogens, including viruses, bacteria and parasites, particularly with the pronounced capacity to defend against SARS-CoV-2, without compromising their oxygen delivery capacity and viability. Conclusion: STIF fosters the immune function of red blood cells and therefore, it may be considered as a nonmedical intervention option for the stronger capacity of red blood cells to combat infectious diseases. Keypoints: STIF fosters the immune response of red blood cells to various pathogens, in particular SARS-CoV-2, via activation of the complement receptor on their membranes. STIF does not compromise the oxygen transport capacity or viability of red blood cells. [ABSTRACT FROM AUTHOR]

Accurate prediction of renewable energy generation acts as a critical role which not only provides short-term power generation in the future, but also facilitates scheduling and pre-configuration of energy storage systems. More importantly, the power generation prediction is of great significance to the demand response management (DRM) of renewable energy to participate in the electricity spot market. Therefore, DRM helps improve the stability and reliability of renewable energy systems. This paper presents a novel prediction-smoothing based methodology to reduce and eliminate the influence caused by the uncertainty of renewable energy output. Firstly, the Whale Optimization Algorithm (WOA) is combined with Long Short-Term Memory (LSTM) to predict short-term wind power output. Then, the Hampel-Butterworth-SG filtering strategy with outlier regression and specific risk band elimination is introduced. After that, according to the short-term output forecast results, the scheduling and pre-configuration scheme of peak-filling type energy storage is developed. Finally, based on the predicted renewable energy output and electricity price, the dynamic changes of demand response (DR) are calculated based on Logistics function, optimistic response and pessimistic response factors. Through extensive case studies, it is demonstrated that the assessment deviations in different scenarios is less than 5%, which are 0.64% for Scenario 1 and 3.64% for Scenario 2, and no additional penalty is required at this time. In addition, the proposed Demand Response Load Adjustment Rate (DRLAR) help compare the differences between predicted and actual DR, which are DRLAR = 0.03% in Scenario 1 and DRLAR = 0.01% in Scenario 2. Users are able to adjust the DR dynamically according to the electricity price to realize the optimal scheduling of their renewable resources. The proposed methodology creates a special connection between DRM and renewable energy prediction, which provides a reliable reference for future work. [ABSTRACT FROM AUTHOR]

I DDX41 i mutations were detected in 0.6% (2/329) adult ALL patients of our cohorts comparable with a recent report.[10] Previous studies showed that I DDX41 i -mutated patients of MNs presented as male predominance and old age. Germline DDX41 mutations define a significant entity within adult MDS/AML patients. We found three variants in I DDX41 i (p.R525H, p.G530D and p.Y259C) and confirmed the germline origin of p.Y259C of I DDX41 i by Sanger sequencing. [Extracted from the article]

Overall survival (OS) (A) and OS censored at the time of transplantation (B) of 24 relapsed pediatric B-ALL patients with mean VAF >=20% versus patients with mean VAF < 20% at relapse. Moreover, in 18 patients with remission-relapse pairs, VAF was significantly increased in relapsed patients, indicating delta VAF can be used to evaluate patients for recurrence ( I p i < 0.001) (Figure 2B). (C) Comparison of VAFs at three different relapse stages in 24 relapsed patients, including very early, early and late relapse stage. [Extracted from the article]

Abstract: Real-time quantitative reverse transcriptase polymerase chain reaction method was established for detecting the expression levels of WT1 gene and WT1+17AA isoforms in 226 acute myeloid leukemia (AML) bone marrow (BM) cells. The results showed that WT1 gene was 2–3 logarithms expressed more in AML BM cells at initial diagnosis or relapse than in normal BM cells (p <0.001), with predominant WT1+17AA isoforms expression (the ratio of WT1+17AA/WT1 more than 0.50). Interestingly the ratio of WT1+17AA/WT1 was statistically higher in relapsed AMLs than in initially diagnosed (p =0.01), speculating that WT1+17AA isoforms might participate in AML relapse. [Copyright &y& Elsevier]

Long non-coding RNAs (lncRNAs) are important in cancer biology. In this study, we analyzed differentially expressed genes in CD34 + hematopoietic cells and identified a novel lncRNA, H22954, which was down-regulated in acute myeloid leukemia (AML) patients. In cultured AML cells and mouse xenograft models, H22954 expression inhibited cell proliferation and tumor growth, respectively. Bioinformatic analysis and RNA antisense purification assay indicated that H22954 targeted the 3' untranslated region of the BCL2 gene. In luciferase assays, H22954 expression inhibited BCL2 expression. In transfected K562 cells and mouse xenograft tumors, H22954 overexpression reduced BCL-2 protein levels and promoted cell death. In AML patients, H22954 expression inversely correlated with BCL-2 protein levels in bone marrow cells, blast cell numbers and disease prognosis. These results indicate that H22954 is a novel regulator of BCL-2 and that reduced H22954 expression may play an important role in the pathogenesis of AML. [ABSTRACT FROM AUTHOR]

Isochromosome 20q- (i(20q-)), as a rare reproducible chromosomal anomaly formed on the basis of 20q-, has not been commonly reported. Due to the rarity of this karyotypic anomaly, the bone marrow morphological characteristics of the patients with i(20q-) have not been clarified until now. In this study, the bone marrow cell morphology from MDS patients with isolated i(20q-), isolated 20q-, and normal karyotype was retrospectively compared and statistically analyzed. The results indicated that the isolated i(20q-) was mostly detected in MDS-MLD patients. The frequency and proportion dysplasia of cytoplasmic vacuolization in erythoid cells and small or unusually large size in myeloid cells of isolated i(20q-) MDS patients were significantly higher than those of normal karyotype MDS patients respectively (P < 0.05); the frequency and proportion dysplasia of decreased granules/agranularity in myeloid cells of isolated i(20q-) MDS patients were higher than those of isolated 20q- MDS patients (P < 0.05). The incidence of some specific morphological manifestations, such as deeply lobulated and hyperlobulated megakaryocytes and hypogranular and vacuolized eosinophils, may be an important morphological implication for the anomaly of isolated i(20q-). These morphological features of dysplasia may be helpful in distinguishing MDS with isolated i(20q-) from those with isolated 20q- and normal karyotype. [ABSTRACT FROM AUTHOR]

Acute graft-versus-host disease (aGvHD) is considered a result of "cytokine storm." Targeted therapeutic interventions on cytokines via ubiquitination regulatory pathways may provide a potential approach for aGvHD treatment. Ubiquitin-specific peptidase 11 (USP11) has been reported to play key roles in a variety of physiopathological processes by regulating the stability and function of several vital protein molecules. However, its role in aGvHD remains unclear. In this study, we identified USP11 was associated with aGvHD in patients. In the aGvHD mouse model, the colon and liver were more seriously affected in recipient mice who received USP11 wt bone marrow (BM) cells and eased after the donor was treated with a USP11 inhibitor or received USP11 ko BM cells. In mouse models, IL-6 was identified as a major effecter in accelerating aGvHD induced by USP11. In the cell model, IL-6 mRNA transcript was affected by USP11. In addition, USP11 also inhibited IL-6 degradation by affecting IL-6 ubiquitination. Furthermore, the positive correlation between USP11 and IL-6 was confirmed in the GvHD patients' samples. Collectively, all results indicated that USP11 played a critical role in the onset and progression of aGvHD. USP11 might be a potential target for aGvHD treatment. [Display omitted] [ABSTRACT FROM AUTHOR]

The article discusses research which investigated plant homeodomain (PHD)-like finger protein 6 (PHF6) gene mutations in patients with chronic myeloid leukemia in blast crisis (CML-BC). Topics explored include the implication of PHF6 in various cellular functions, the chromosomal changes that contribute to the progression of CML to BC, and the analysis of the positive PHF6 mutations recorded following the genomic DNA sequencing performed.

Background BCL11A encodes a C2H2 type zinc-finger protein. During normal haematopoietic cell differentiation BCL11A expression is down-regulated. Data in mice suggest up-regulation of BCL11A is involved in the pathogenesis of myeloid leukaemias. BCL11A expression in persons with acute myeloid leukaemia (AML) is not systematically studied. Objective Interrogate associations between BCL11A expression at diagnosis and clinical and laboratory valuables and outcomes in newly-diagnosed persons with AML. Methods We determined BCL11A mRNA levels in bone marrow and blood mononuclear cells in 292 consecutive newly-diagnosed subjects with AML by reverse transcript and real-time polymerase chain reaction. Data were compared to mRNA levels in bone marrow cells of normals. Results Subjects with BCL11A transcript levels at diagnosis exceeding the median value of 2.434 (±3.423 SD; 25th–75th inter-quartile range, 1.33–4.29) had higher WBC levels, a greater proportion of bone marrow myeloblasts, were more likely to be FAB M 0 subtype, less likely to be FAB M 3 subtype, more likely to be in the intermediate cytogenetic risk cohort, less likely to have a complex karyotype and more likely to have DNMT3A R882 and FLT3 -ITD mutations than subjects with transcript levels below the median value. In 89 subjects receiving conventional induction chemotherapy the complete remission rate was 54% (95% confidence interval [CI]; 33, 75%) in the lower BCL11A cohort and 65% (45, 85%; P = 0.26) in the higher BCL11A cohort. 3 year survival was 33% (2, 65%) in the lower BCL11A cohort and 15% (0, 39%; P = 0.35) in the high BCL11A cohort. Conclusion BCL11A transcript levels at diagnosis was significantly associated with several clinical and laboratory variables. There were also non-significant associations with complete remission rate and survival. These data suggest a possible role for BCL11A expression in AML biology. [ABSTRACT FROM AUTHOR]

This retrospective study was designed to evaluate the clinical and laboratory behaviors of chromosome 3 abnormalities by analyzing the morphological, cytogenetic, and follow-up data from 300 patients of various hematological malignancies with chromosome 3 abnormalities. From the results, trisomy 3, translocation (3q), and del(3) were the abnormal types most frequently observed (>10%) among the chromosome 3 abnormalities. In hematological malignancies, chromosome 3 abnormalities were most frequently seen in the patients with acute myeloid leukemia (AML) (24.7%) and myelodysplastic syndrome (MDS) (16%), followed by those with acute lymphocytic leukemia (ALL) (13.7%) and multiple myeloma (MM) (12.7%). In this series, genomic losses were the most frequent genetic abnormalities in AML, ALL, and hybrid acute leukemia (HAL) patients, whereas structural rearrangements were frequently seen in chronic myeloid leukemia (CML) and MDS patients, and genomic gains in MM, lymphoma and chronic lymphocytic leukemia (CLL) patients. Chromosome 3 abnormalities mainly occurred as a component of a complex abnormality (251/300) rather than a sole one (14/300). Survival analysis demonstrated a statistical difference between the patients with trisomy 3, who had a better prognosis, and patients with del(3), who had a worse prognosis in this series ( P < 0.05). Abnormalities in chromosome 3 may imply an unfavorable outcome in CML and ALL. [ABSTRACT FROM AUTHOR]

Background: This study was conducted to determine the antineoplastic activities of 5-aza-2′-deoxycytidine (decitabine; DAC) and all-trans retinoic acid (ATRA), administered either alone or in combination, on in vitro cultured SHI-1 cells as well as their effects on the expression of the tumor suppressor gene p16INK4a (p16) and the retinoic acid receptor (RAR)-β. Methods: Cellgrowth inhibition, differentiation and apoptosis were determined in SHI-1 cells treated with DAC and/or ATRA, and the combination index of the two compounds was calculated. Methylation of the p16 and RAR-β genes in SHI-1 cells was detected by methylation-specific polymerase chain reaction (PCR). Real-time quantitative reverse transcriptase PCR was used to detect mRNA expression of the p16 and RAR-β genes, and Western blot analysis was performed for protein expression. Results: The drug combination had a synergistic effect on growth inhibition, differentiation and apoptosis of SHI-1 cells, and the effects of DAC and ATRA weredependent on time. DAC, either alone or in combination with ATRA, induced demethylation of the genes p16 and RAR-β, whereas ATRA alone had no effect on methylation. The RAR-β gene was reexpressed following DAC-ATRA combination treatment, and both agents had no effect on p16 expression. Conclusion: The results revealed that DAC used in combination with ATRA has significant clinical potential in the treatment of acute monocytic leukemia. © 2014 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Aims: The object of the present study was to develop a novel multiplex reverse transcription-polymerase chain reaction (RT-PCR)-based assay to detect four common recurrent chromosomal translocations t(9;22)(q34;q11), t(15;17)(q22;q12), t(8;21)(q22;q22), and t(1;19)(q23;q13). Results: This novel multiplex RT-PCR method can specifically detect these six positive plasmids with known transcripts, and the sensitivities were up to 10 copies. In the diagnosis of clinical specimens, four chromosomal translocations, including seven transcripts, were actually specifically detected. Conclusions: This newly developed multiplex RT-PCR system can be used to detect these four recurrent chromosomal translocations specifically and sensitively. It is novel, accurate, and time and cost effective in the detection of leukemia-specific chromosomal translocations. Assays with this new strategy will have a potential in detecting other chromosomal translocations in leukemias. [ABSTRACT FROM AUTHOR]

The myelodysplastic syndromes (MDS) comprise a heterogeneous group of malignant neoplasms with distinctive clinicopathological features. Currently, there is no specific approach for the treatment of MDS. Here, we report that bortezomib (BTZ), a proteasome inhibitor that has been used to treat plasma cell myeloma, induced G2/M phase cycle arrest in the MDS cell line SKM-1 through upregulation of Wee1, a negative regulator of G2/M phase transition. Treatment by BTZ led to reduced SKM-1 cell viability as well as increased apoptosis and autophagy. The BTZ-induced cell death was associated with reduced expression of p-ERK. To elucidate the implications of downregulation of p-ERK, we established the BTZ resistant cell line SKM-1R. Our data show that resistance to BTZ-induced apoptosis could be reversed by the MEK inhibitors U0126 or PD98059. Our results suggest that MAPK pathway may play an important role in mediating BTZ resistance. [ABSTRACT FROM AUTHOR]

The role of tissue inhibitor of metalloproteinase-2 (TIMP-2) in extramedullary infiltration of acute leukemia is unclear. We demonstrated in our previous study that the up-regulation of TIMP-2 promoted SHI-1 cell invasion in vitro. We investigated in the present study whether TIMP-2 would have the same effect in vivo. A retroviral vector carrying human TIMP-2 cDNA was constructed and transfected into SHI-1 cells. Three subclone cells (S1, S2 and S3) that highly expressed TIMP-2 were selected to establish nude mouse models of acute leukemia. Times of leukemic onset in mice of S1, S2 and S3 groups were all earlier than that of the SHI-1 group, whereas the survival times of S1, S2 and S3 groups were all shorter than that of the SHI-1 group ( p < 0.05). Histopathological results demonstrated severe leukemic infiltration in numerous organs in each group. Reverse transcription polymerase chain reaction (RT-PCR) assay showed that several organs expressed the MLL/F6 fusion gene. Moreover, the numbers of organs infiltrated by leukemic cells in S1, S2 and S3 groups were more than those in the SHI-1 group ( p < 0.05). Up-regulating TIMP-2 expression enhanced SHI-1 cell invasion in nude mice and resulted in more severe leukemia infiltration. This phenomenon suggests that targeted therapy with TIMP-2 for acute leukemia should be performed with prudence. [ABSTRACT FROM AUTHOR]

This study aimed to determine the clinical characteristics and prognostic significance of the meningioma 1 ( MN1) gene and MN1-associated microRNA expression in Chinese adult de novo acute myeloid leukemia (AML) patients. The expression level of MN1, microRNA- 20 ( miR- 20a), and microRNA- 181b ( miR- 181b) in bone marrow mononuclear cells was measured in 158 newly diagnosed AML patients and 20 cases of normal healthy donors by real-time quantitative reverse transcriptase polymerase chain reaction. All AML patients significantly overexpressed MN1 at the level of 0.01983 ( P < 0.001) compared with normal controls. High MN1 expression was associated with spleen involvement ( P = 0.037), NPM1 wild type ( P = 0.001), lower miR- 20a expression levels ( P = 0.015), and higher miR- 181b expression levels ( P = 0.035). MiR- 20a ( P = 0.029) and miR- 181b ( P = 0.017) overexpressed in the bone marrow cells of patients with certain subtypes of AML compared with healthy donors. High MN1 expressers had lower complete remission (CR) rates and shorter overall survival (OS) within the Southwest Oncology Group classification. In multivariable models, high MN1 expression was associated with worse CR rates ( P = 0.01), relapse-free survival (RFS; P = 0.02), and OS ( P = 0.02); high miR- 20a expression was associated with higher CR rates ( P = 0.008) and longer OS ( P = 0.04), whereas high miR- 181b expression was associated with lower CR rates ( P = 0.03), and shorter RFS ( P = 0.045) and OS ( P = 0.017). High MN1 expression confers worse prognosis in Chinese adult patients with de novo AML. MN1 gene and MN1-associated microRNAs provide clinical prognosis of AML patients and may refine their molecular risk classification. [ABSTRACT FROM AUTHOR]

The IK6 isoform plays an important role in Philadelphia chromosome-positive adult acute lymphoblastic leukemia (Ph + ALL). This study was designed to monitor the expression of the IK6 isoform with associated cytogenetic and molecular abnormalities. The IK6 isoform, cytogenetic and molecular abnormalities were detected in 100 Chinese patients with de novo Ph+ adult ALL. Expression levels of the IK6 isoform and BCR-ABL1 transcripts were monitored during treatment. BCR-ABL1 mutation was identified in 45 paired samples. Strong correlations were found between the expression status of the IK6 isoform and blast cells, additional cytogenetic abnormalities, BCR-ABL1 transcripts, increased risk of relapse, shorter relapse-free survival and overall survival at diagnosis. Higher frequencies of single IK6 expression and ABL mutation, including the types and shifts thereof, were confirmed in relapsed patients. Furthermore, expression of the IK6 isoform was dynamically consistent with BCR-ABL1 transcript levels during treatment in the single expression group, whereas no such correlation was observed in the co-expression group. The expression pattern of the IK6 isoform was altered in three patients from the co-expression group. The findings of this study in Chinese patients with Ph+ adult ALL exhibit some discrepancies with data reported in other countries, thereby enhancing current knowledge on the therapeutic response and prognosis of this disease. [ABSTRACT FROM AUTHOR]

Insulin-like growth factor--binding protein 7 (IGFBP7) has been identified as a tumor suppressor in solid tumors. In acute leukemia, the role of IGFBP7 is largely unknown. The authors used quantitative reverse transcriptase--polymerase chain reaction (qRT-PCR) to investigate the expression level of IGFBP7 gene in bone marrow (BM) specimen from 66 children with acute myeloid leukemia (AML) at different stages and in 30 nonleukemia patients as control. Furthermore, U937 cells were transfected with siRNA-2 of IGFBP7 (as U937R) for 24 hours. Coculture experiment was performed to explore the impact of IGFBP7 gene in U937 cell adhesion, invasion, and migration in existing ECV304 cells, which mimicked the interaction between AML cells and endothelial cells. IGFBP7 expression at the initial diagnosed stage and relapse of AML was significantly higher than that of control ( P < .001). The viable cell percentage in transfected cell was significantly decreased by 42%% compared with control groups ( P < .01). The percentage for U937R cells adherent to ECV304 cells was significantly lower than the control groups ( P < .01). Matrigel study to quantify the invasive potential showed that U937R migrated to the lower chamber were significantly less than those in the parental control groups ( P < .01). In summary, IGFBP7 aberrantly overexpressed in majority of AML at diagnosis and upon relapsed, but not at remission stage. IGFBP7 plays a positive contributing role in the interaction between leukemia cells and microenvironment, which may promote the leukemic cells'' adhesion, invasion, and migration. [ABSTRACT FROM AUTHOR]

Abstract: Killer cell immunoglobulinlike receptors (KIRs) are a diverse family of receptors on natural killer cells. The KIR2DS4-deleted variant differs from the normal KIR2DS4 sequence by a 22-bp deletion in exon 5, which causes a frame shift, yielding a truncated KIR2DS4 protein with loss of the transmembrane and cytoplasmic domains of the full-length KIR2DS4 protein. A sequence-based testing and TOPO TA cloning system identifying and distinguishing alleles of the KIR2DS4 gene was established. The method was applied to 150 Chinese Han individuals: 75 patients who received T-depleted hematopoietic stem cells transplantation and their unrelated donors, establishing frequencies of 2DS4 allele and KIR gene within the local population. A majority (139) of the 150 samples (92.7%) were positive for KIR2DS4. The ratio of deleted to nondeleted versions of KIR2DS4 was approximately 1:2. Three KIR2DS4 novel alleles were identified. Forty-four percent individuals carried two group A haplotypes. The 17 KIR gene loci were found in 21.3%–100% in the population. Our findings suggest that the Chinese Han population is distinct in KIR gene frequencies and 2DS4 allele frequencies in comparison with some other populations. [Copyright &y& Elsevier]

Abstract: TNFR-associated factors (TRAFs) participate in diverse biological processes, such as adaptive and innate immunity, stress response, and bone metabolism. We report that all TRAFs except TRAF3 are expressed at mRNA and protein levels in B cell-derived Hodgkin''s lymphoma cell lines (L428 and KM-H2). Both the classical (p50–RelA) and the alternative NF-κB activity (p52–RelB) are sustained in L428 and KM-H2 cells. A successful depletion of TRAF1 protein expression by means of RNA interference abrogates the anti-apoptosis activity in L428 cells. The TRAF1-deficiency reduces the classical NF-κB activity but not the alternative activity. The expression of the NF-κB targeting genes, such as ICAM-1, c-Flip, and Cyclin D1, is suppressed in the TRAF1-depleted cells. On the other hand, CD30 signaling upregulates the TRAF1 expression while reducing the expression of TRAF2 and TRAF5. Importantly, the CD30-induced alternative NF-κB activation is inhibited by the depletion of the TRAF1 expression. We also demonstrate that the phosphorylation of the extracellular signal-regulated kinase (ERK) upon CD30 stimulation in Hodgkin''s lymphoma cells is independent of TRAF1 expression. Our data shed new light on the function of TRAF1 in B cell-derived lymphoma cells. We conclude that TRAF1 is an important molecule mediating both the CD30 signaling-dependent and independent NF-κB activation, which prevents the lymphoma cells from spontaneous and induced apoptosis. [Copyright &y& Elsevier]

A letter to the editor is presented regarding a study on the calreticulin (CALR) and additional sex combs like 1 (ASXL1) mutation in patients with essential thrombocythemia (ET).

A letter to the editor is presented which discusses a study showing that DNA methyltransferases (DNM)T3A has nothing to do with leukemogenesis of chronic myeloid leukemia (CML) and the progression from CML to chronic phase (CP) and from CML to blast crisis (BC).