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Objectives To compare the clinical significance of SF3B1/DNMT3A Comutations with SF3B1 or DNMT3A mutation alone in myelodysplastic syndrome (MDS) and clonal cytopenia of undetermined significance (CCUS). Methods We identified and compared 31 patients with only DNMT3A mutation, 48 patients with only SF3B1 mutation, and 16 patients with only SF3B1/DNMT3A comutations. Results SF3B1/DNMT3A comutations were found to be more common in MDS, whereas DNMT3A mutation alone was more common in CCUS. The patients with SF3B1/DNMT3A comutations were less likely to have poor cytogenetics than patients with DNMT3A mutation alone. Patients with SF3B1/DNMT3A comutations showed significantly longer median survival time and better overall survival than patients with DNMT3A mutation alone. Conclusions Patients with SF3B1/DNMT3A comutations appear to have better clinical outcomes than patients with isolated DNMT3A mutation. These findings suggest that the favorable prognosis of SF3B1 mutation in is not abrogated by the concurrent presence of a DNMT3A mutation.

Lipopolysaccharide and colanic acid are important forms of exopolysaccharides located on the cell surface of Escherichia coli, but their interrelation with the cell stress response is not well understood. In this study, nine mutant strains with different structures of lipopolysaccharide were constructed from E. coli MG1655 by deletion of a single gene or multiple genes. All mutant strains did not produce colanic acid when grown in LB medium, but six of them could produce colanic acid when grown either in M9 medium in which glucose is the sole carbon source or in LB medium supplemented with glucose. The results indicate that colanic acid production in E. coli is dependent on both lipopolysaccharide structure and glucose availability. However, transcriptional analysis showed that 20 genes related to the colanic acid biosynthesis and the key gene rcsA in the Rcs system were all transcriptionally up-regulated in all of the nine mutant strains no matter they were grown in M9 or LB medium. This suggests that the availability of some nucleotide-sugar precursors shared by the biosynthesis of lipopolysaccharide and colanic acid might play a major role in colanic acid production in E. coli. Lipopolysaccharide pathway might have a huge priority to colanic acid pathway to use the common precursors; therefore, the colanic acid is not produced in MG1655 and the nine mutants when grown in LB medium. In the six mutant strains that can produce colanic acid in the glucose rich media, the common precursors might be abundant because they were not needed for synthesizing the mutant lipopolysaccharide.

Wild-type Escherichia coli usually does not accumulate l-threonine, but E. coli strain TWF001 could produce 30.35 g/L l-threonine after 23-H fed-batch fermentation. To understand the mechanism for the high yield of l-threonine production in TWF001, transcriptomic analyses of the TWF001 cell samples collected at the logarithmic and stationary phases were performed, using the wild-type E. coli strain W3110 as the control. Compared with W3110, 1739 and 2361 genes were differentially transcribed in the logarithmic and stationary phases, respectively. Most genes related to the biosynthesis of l-threonine were significantly upregulated. Some key genes related to the NAD(P)H regeneration were upregulated. Many genes relevant to glycolysis and TCA cycle were downregulated. The key genes involved in the l-threonine degradation were downregulated. The gene rhtA encoding the l-threonine exporter was upregulated, whereas the genes sstT and tdcC encoding the l-threonine importer were downregulated. The upregulated genes in the glutamate pathway might form an amino-providing loop, which is beneficial for the high yield of l-threonine production. Many genes encoding the 30S and 50S subunits of ribosomes were also upregulated. The findings are useful for gene engineering to increase l-threonine production in E. coli.

(R)-Selective ω-transaminase (ω-TA) is a key enzyme for the asymmetric reductive amination of carbonyl compounds to produce chiral amines which are essential parts of many therapeutic compounds. However, its practical industrial applications are hindered by the low catalytic efficiency and poor thermostability of naturally occurring enzymes. In this work, we report the molecular modification of (R)-selective ω-TA from Aspergillus terreus (AtTA) to allow asymmetric reductive amination of 4-hydroxy-2-butanone, producing (R)-3-amino-1-butanol. Based on substrate docking analysis, 4 residues in the substrate tunnel and binding pocket of AtTA were selected as mutation hotspots. The screening procedure was facilitated by the construction of a “small-intelligent” library and the use of thin-layer chromatography for preliminary screening. The resulting mutant AtTA-M5 exhibited a 9.6-fold higher kcat/Km value and 9.4 °C higher $$ {\mathrm{T}}_{1/2}^{10\min } $$ than that of wild-type AtTA. Furthermore, the conversion of 20 and 50 g L−1 4-hydroxy-2-butanone by AtTA-M5 reached 90.8% and 79.1%, suggesting significant potential for production of (R)-3-amino-1-butanol. Under the same conditions, wild-type AtTA achieved less than 5% conversion. Moreover, the key mutation (S215P in AtTA) was validated in 7 other (R)-selective ω-TAs, indicating its general applicability in improving the catalytic efficiency of homologous (R)-selective ω-TAs.

In recent years, the prevalence of depression has been increasing, causing a serious global burden of disease. Previous studies have proved that low-grade inflammation is involved in the process of depression. To explore the role of inflammation in the pathogenesis of senile depression, neutrophil to lymphocyte ratio (NLR) was used in this study, as a biological marker for non-specific inflammation. The present study was conducted among elderly adults from a randomly selected Community in Yangpu District, Shanghai from November 1 to December 30, 2017. 665 participants were included in the present study, including 276 males (median age was 68 years old) and 389 females (median age was 67 years old). NLR was calculated based on results of hematology examination. Depressive symptoms were assessed using the simplified version of Geriatric Depression Scale (GDS), the participant with total score greater than five points was considered to be depressed. The association between NLR and depression was evaluated separately for men and women. The prevalence of depression was 35.1% for men and 34.4% for women. No associations were found between NLR and depression among males. Nevertheless, compared with normal women, NLR was significantly increased in women with depressive symptoms (P = 0.020). Logistic regression analysis showed that NLR was independently associated with geriatric depression in women. The odds ratios (ORs) of depression was 2.152 (1.095, 4.227; P = 0.026) for the fourth compared with the first quartile of NLR. In conclusion, this study showed increased NLR was significantly associated with geriatric depression in women, but not men.

White matter lesions are common when global cerebral ischemia (GCI) occurs in the elderly, and cause damage to neurological and psychological functions. Remyelination often fails because of the limited recruitment of oligodendrocyte progenitor cells (OPCs) to the demyelinated site or the inefficient differentiation of OPCs to mature oligodendrocytes (OLs). The activation of microglia, the most important immune cells in the central nervous system, and subsequent inflammation have been implicated in myelination repair disorder. Little is known about the role of the Fractalkine/CX3CR1 signaling pathway, the key regulator of microglia activation, on myelin in microglia. In this study, a GCI animal model was generated through bilateral common carotid artery occlusion to induce ischemic inflammation and white matter damage; then, we downregulated CX3CR1 by intracerebroventricular administration of neutralizing antibody anti-FKR. Downregulation of CX3CR1 significantly reversed the depression-like behavior and cognitive impairment in GCI mice. Activation of microglia was inhibited, and the peripheral inflammatory responses were also ameliorated as revealed by decreased serum levels of IL-1β, IL-6 and TNF-α. CX3CR1 block substantially reversed demyelination in striatum, cortex and hippocampus and promoted differentiation and maturation of OPCs into mature OLs in the hippocampus. No effect was found on myelin in the corpus callosum. Besides, hippocampal neurons were protected by anti-FKR treatment after GCI. Collectively, our data demonstrated that downregulating of the Fractalkine/CX3CR1 signaling pathway had an anti-depressant and cognition-improvement effect by inhibiting microglia activation, promoting OPCs maturation and remyelination.

Background Several studies have recently found that oxidative stress plays a pivotal role in the pathogenesis of traumatic brain injury (TBI) and may represent a target in TBI treatment. Hydrogen-rich water was recently shown to exert neuroprotective effects in various neurological diseases through its antioxidant properties. However, the mechanisms underlying its effects in TBI are not clearly understood. The purpose of our study was to evaluate the neuroprotective role of hydrogen-rich water in rats with TBI and to elucidate the possible mechanisms underlying its effects. Materials and methods The TBI model was constructed according to the modified Feeney weight-drop method. In part 1 of the experiment, we measured oxidative stress levels by observing the changes in catalase (CAT), glutathione peroxidase (GPx), and malondialdehyde (MDA) expressions. We also evaluated nuclear factor erythroid 2–related factor 2 (Nrf2) levels to determine the role of the protein in the neuroprotective effects against TBI. In part 2, we verified the neuroprotective effects of hydrogen-rich water in TBI and observed its effects on Nrf2. All the experimental rats were divided into sham group, TBI group, and TBI + hydrogen-rich water-treated (TBI + HW) group. We randomly chose 20 rats from each group and recorded their 7-d survival rates. Modified neurological severity scores were recorded from an additional six rats per group, which were then sacrificed 24 h after testing. Spectrophotometry was used to measure GPx, CAT, and MDA levels, whereas western blotting, reverse transcription polymerase chain reaction, and immunohistochemistry were used to measure the expression of Nrf2 and downstream factors like heme oxygenase 1 (HO-1) and NAD(P)H quinone oxidoreductase 1 (NQO1). Results GPx and CAT activity was significantly decreased, and MDA content was increased in the TBI group compared with the sham group at 6 h after TBI. MDA content peaked at 24 h after TBI. Nrf2 nucleoprotein levels were upregulated in the TBI group compared with the sham group and peaked at 24 h after TBI; however, no significant changes in Nrf2 mRNA levels were noted after TBI. Hydrogen-rich water administration significantly increased 7-d survival rates, reduced neurologic deficits, and lowered intracellular oxidative stress levels. Moreover, hydrogen-rich water caused Nrf2 to enter the cell nucleus, which resulted in increases in the expression of downstream factors such as HO-1 and NQO1. Conclusions Our results indicate that hydrogen-rich water has neuroprotective effects against TBI by reducing oxidative stress and activating the Nrf2 pathway.

In metabolic engineering, unbalanced microbial carbon distribution has long blocked the further improvement in yield and productivity of high-volume natural metabolites. Current studies mostly focus on regulating desired biosynthetic pathways, whereas few strategies are available to maximize L-threonine efficiently. Here, we present a strategy to guarantee the supply of reduced cofactors and actualize L-threonine maximization by regulating cellular carbon distribution in central metabolic pathways. A thermal switch system was designed and applied to divide the whole fermentation process into two stages: growth and production. This system could rebalance carbon substrates between pyruvate and oxaloacetate by controlling the heterogenous expression of pyruvate carboxylase and oxaloacetate decarboxylation that responds to temperature. The system was tested in an L-threonine producer Escherichia coli TWF001, and the resulting strain TWF106/pFT24rp overproduced L-threonine from glucose with 111.78% molar yield. The thermal switch system was then employed to switch off the L-alanine synthesis pathway, resulting in the highest L-threonine yield of 124.03%, which exceeds the best reported yield (87.88%) and the maximum available theoretical value of L-threonine production (122.47%). This inducer-free genetic circuit design can be also developed for other biosynthetic pathways to increase product conversion rates and shorten production cycles.

L-Valine is one of the three branched-chain amino acids (valine, leucine, and isoleucine) essential for animal health and important in metabolism; therefore, it is widely added in the products of food, medicine, and feed. L-Valine is predominantly produced through microbial fermentation, and the production efficiency largely depends on the quality of microorganisms. In recent years, continuing efforts have been made in revealing the mechanisms and regulation of L-valine biosynthesis in Corynebacterium glutamicum, the most utilitarian bacterium for amino acid production. Metabolic engineering based on the metabolic biosynthesis and regulation of L-valine provides an effective alternative to the traditional breeding for strain development. Industrially competitive L-valine-producing C. glutamicum strains have been constructed by genetically defined metabolic engineering. This article reviews the global metabolic and regulatory networks responsible for L-valine biosynthesis, the molecular mechanisms of regulation, and the strategies employed in C. glutamicum strain engineering.

Hairy cell leukemia (HCL) is a rare and indolent mature B cell neoplasm. Most patients with HCL have excellent response to purine analogs and BRAF inhibitors. Therefore, it is clinically important to identify and differentiate HCL from other B cell neoplasms. HCL is predominantly found in the peripheral blood, bone marrow, or spleen. HCL presenting initially or solely in unusual sites is extremely rare (approximately ten or fewer). We hereby report a unique case of HCL presenting as vertebral osteolytic and epidural mass lesions without bone marrow, spleen, or peripheral blood involvement. The patient was a 53-year-old male with acute and chronic radicular pain down the posterior aspect of bilateral thighs. Imaging study with MRI demonstrated a L4 posterior vertebral osteolytic mass lesion, extending into the spinal canal, and a second right sacral S1 lesion extending into the sacral iliac joint. These two noncontiguous lesions were thought to be consistent with metastatic cancer or sarcoma, and the specimen was submitted to non-hematopoietic pathologist for diagnosis without flow cytometry. Initial immunohistochemical stains were ordered for solid tumor markers, which were all negative. As part of the workup, immunostain for BRAF V600E (VE1) was ordered for melanoma, and CD138 and cyclin D1 for myeloma, which returned positive and suggested the possibility of hairy cell leukemia. Additional workup for hairy cell leukemia confirmed the diagnosis. HCL typically presents with peripheral blood, bone marrow, or spleen involvement, while mass lesions in other sites are rare. High index of suspicion is essential for the correct diagnosis of HCL in mass lesions at unusual sites. In the absence of initial hematopathological workup and flow cytometry, HCL presenting as solid mass in unusual locations is misdiagnosed most likely as marginal zone B cell lymphoma (mucosa-associated lymphoid tissue (MALT) lymphoma). The expression of cyclin D1, TRAP, annexin A1, and BRAF V600E mutation would confirm the diagnosis of HCL. The main differentials include MALT lymphoma, mantle cell lymphoma (MCL), and hairy cell leukemia variant (HCLv). This case report will be interesting for the clinicians and pathologists alike and broaden the spectrum of clinical presentation of lymphomatous HCL.

BACKGROUND Breast cancer and its treatment provoke a series of emotional changes in patients during their breast cancer journeys. Social support is critical in helping women cope with their negative emotional responses. However, few studies have described the experiences of women with breast cancer within the Chinese context. This qualitative study explored the experiences of Chinese women with breast cancer regarding social support. MATERIAL AND METHODS This qualitative study was based on constructivism epistemology. Chinese women with breast cancer were invited to interview between June and August 2016. Purposive sampling was used, and the women were recruited until data saturation was reached (n=25). RESULTS We found that all participants expressed the importance of social support during their breast cancer journeys. The thematic framework of social support was identified, which included the following 4 interrelated themes: providers, types of support, outcomes of support, and challenges for care. CONCLUSIONS Social support is important in the health care of women living with breast cancer. Healthcare professionals should provide more individualized support and care to women during this vulnerable period.

Inflammation and oxidative stress play a crucial role in the development of diabetic cardiomyopathy (DCM). We previously had synthesized an Aza resveratrol–chalcone derivative 6b, of which effectively suppressing lipopolysaccharide (LPS)‐induced inflammatory response in macrophages. This study aimed to investigate the potential protective effect of 6b on DCM and underlying mechanism. In H9c2 myocardial cells, 6b potently decreased high glucose (HG)‐induced cell fibrosis, hypertrophy and apoptosis, alleviating inflammatory response and oxidant stress. In STZ‐induced type 1 diabetic mice (STZ‐DM1), orally administration with 6b for 16 weeks significantly attenuated cardiac hypertrophy, apoptosis and fibrosis. The expression of inflammatory cytokines and oxidative stress biomarkers was also suppressed by 6b distinctly, without affecting blood glucose and body weight. The anti‐inflammatory and antioxidative activities of 6b were mechanistic associated with nuclear factor‐kappa B (NF‐κB) nucleus entry blockage and Nrf2 activation both in vitro and in vivo. The results indicated that 6b can be a promising cardioprotective agent in treatment of DCM via inhibiting inflammation and alleviating oxidative stress. This study also validated the important role of NF‐κB and Nrf2 taken in the pathogenesis of DCM, which could be therapeutic targets for diabetic comorbidities.

Graphical abstract Image, graphical abstract, Posttransplant lymphoproliferative disorders (PTLDs) of T-cell orgin are rare biologically heterogeneous diseases of mature lymphoid cells manifesting in immunosuppressed patients. Only a few cases of mycosis fungoides diagnosed post allogeneic hematopoietic cell transplant (alloHSCT) have been described so far. We present a patient with myelodysplastic syndrome (MDS) post matched unrelated donor alloHSCT who was on long-term immunosuppressive therapy due to graft versus host disease. Three years after an alloHSCT, she developed generalized erythroderma and peripheral blood lymphocytosis. Both skin biopsy and peripheral blood flow cytometry revealed atypical CD4+ T-cell population consistent with diagnosis of Sezary syndrome. Chimerism studies revealed 100% donor engraftment. Therapy with extracorporeal photopheresis resulted in complete response in blood and skin.

BACKGROUND The diagnosis and treatment of breast cancer can provoke a series of negative emotional changes in patients, further affecting their quality of life. It has been shown that patients with higher resilience have better quality of life. Social support systems are important protective factors that are necessary for the process of resilience to occur. Hence, this study aimed to investigate the role of social support in the relationship between resilience and quality of life among Chinese patients with breast cancer. MATERIAL AND METHODS A demographic-disease survey, the Chinese version of the Connor-Davidson Resilience Scale 25, Medical Outcomes Study Social Support Survey, and Functional Assessment of Cancer Therapy Breast Cancer Version 3 were used to interview 98 patients with breast cancer from a teaching hospital in Chongqing, China. Data analysis was performed by descriptive statistics, independent-sample t test, one-way ANOVA, and regression analyses. RESULTS The mean scores of resilience, social support, and quality of life were 54.68, 61.73, and 80.74 respectively, which were in the moderate range. Participants with stronger social support had higher resilience and better quality of life. Social support played a partial mediator role in the relationship between resilience and quality of life. The mediation effect ratio was 28.0%. CONCLUSIONS Social support is essential for the development of resilience and the improvement of quality of life in Chinese patients with breast cancer. Health professionals should provide appropriate guidelines to help patients seek effective support and enhance their resilience to improve their quality of life after breast cancer.

The high-temperature oxidation of Hf-modified NiAl alloy at 950 °C results in the formation of γ-Al2O3 and θ-Al2O3. Exclusive formation of HfO2 particles is observed at the adherent γ-Al2O3/NiAl interface. By contrast, the θ-Al2O3/NiAl interface oxidation results in multi-layered oxide growth consisting of alternate thin and thick layers of θ-Al2O3 and NiAl2O4 along with dispersed HfO2 particles in the oxide layers. This difference is elucidated by first-principle calculations showing that the θ-Al2O3 lattice is more favorable than γ-Al2O3 to host Hf and Ni atoms, thereby resulting in the growth of mixed oxides at the θ-Al2O3/NiAl interface.

In order to better achieve efficiently simultaneous desulfurization and denitrification/S0 recovery of wastewater, the intervention of sulfur oxidizing bacteria (SOB) and denitrifying bacteria (DNB) was employed to avoid the collapse critical points (the dramatically decrease of S/N removal efficiency) under the fluctuated load. With the assistance of DNB and SOB, collapse critical point of trickling filter (TF) was delayed from the P8 (105–114 d) to P10 stage (129–138 d). The treatment efficiency of nitrogen and sulfur was the highest with the S/N ratio of 3:1. The bioaugmentation of DNB and SOB at collapse critical point could effectively regulated collapse situation, which further increased the maximum system utilization/elimination capacity to 4.50 kg S m−3·h−1 and 0.90 kg N m−3·h−1 (increased by 56.89% and 65.56% in comparison to control). High-throughput sequencing analysis indicated that Proteobacteria (average 78.59%) and Bacteroidetes (average 9.30%) were dominant bacteria in the reactor at all stages. As the reaction proceeds, the microbial community was gradually dominated by some functional genera such as Chryseobacterium (average 2.97%), Halothiobacillus (average 22.71%), Rhodanobacter (average 14.02%), Thiobacillus (average 9.01%), Thiomonas (average 16.70%) and Metallibacterium (average 21.63%), which could remove nitrate or sulfide. Both of Principal Component Analysis (PCA) and Canonical Correlation Analysis (CCA) demonstrated the important role of DNB/SOB during the long-term run in the trickling filters (TFs).

Integrated assessment models (IAMs) have been widely used to assess the effect of climatic policy. They are used to combine economic activity with simple climatic systems. Coupling earth system models (ESMs) to a socioeconomic framework for scientific research on climate change impacts is a challenging task. The coupling mechanism for complicated climate information in ESMs and economic activities, particularly the effects of extreme climate events on economic activities of industrial sectors, needs to be further explored. Therefore, we constructed a new IAM named EMRICES-2021 and improved the key coupling technology of damage function by adding an economic impact mechanism of precipitation, and extreme climate events. Considering the dynamic persistence features of climatic impacts on industrial sectors, we embedded the new damage function into the Cobb-Douglas production function of a computable general equilibrium (CGE) model and simulated future climatic losses on industrial economic system. Results show that the new damage function could better simulate the sudden economic impacts of extreme climate events and volatilities of climatic losses than Nordhaus mode. The cumulative indirect losses caused by extreme weather far exceeded direct losses in RCP scenarios. Climatic economic losses on real estate, finance, wholesale and retail trade increased fastest.

Background: It is necessary to systematically evaluate the clinical efficacy and safety of bevacizumab (BEV) combined with 5-fluorouracil + leucovorin + oxaliplatin (FOLFOX) regimen in the treatment of advanced colorectal cancer. Methods: We searched the PubMed et al databases for randomized controlled trials (RCTs) on the BEV combined with the FOLFOX regimen in the treatment of advanced colorectal cancer up to January 20, 2021. The Cochrane Collaborations’ risk of bias tool was used for the quality assessment of included RCTs. Revman5.3 software was used for meta-analysis. Results: Eleven RCTs with a total of 3178 patients with advanced colorectal cancer were included, meta-analysis results showed that the objective response rate (odds ratio [OR] = 3.15, 95% confidence intervals [CI]: 2.25–4.40, P

Background: Multiple myeloma (MM) is a clonal plasma cell neoplasm typically associated with chronic therapy and resultant potential toxicities, including clonal cytopenias, myelodysplastic syndrome (MDS), or therapy-related myeloid neoplasms (tMN). Early identification of myelodysplasia is important for patient management and outcome. Next generation sequencing (NGS) is playing an ever increasing role in this field. Materials and Methods: The retrospective study was approved by Moffitt institutional review board (IRB). We searched our in-house NGS database with ~6000 patients and clinical databases to identify the patients with MM and sustained cytopenia with accompanying NGS data. The NGS results were analyzed for associations with myeloma and myelodysplasia. Results: Of the 196 identified patients identified (Table 1), there were 114 males (58%) and 82 females (42%) with a median age of 68 years. Eighty-four myeloma patients with cytopenia (43%) were found to have one or more somatic mutations and 112 patients (57%) showed no mutations. The most frequently mutated genes are as following: TP53 (12%), DNMT3A (8%), TET2 (6%), ASXL1 (5%), KRAS (5%), ETV6 (3%), RUNX1 (2%), CUX1 (2%), BCOR (2%), SF3B1 (2%), ZRSR2 (2%), EZH2 (2%), IDH2 (2%), SRSF2 (2%), and BRAF (1%). We divided the patients into four groups according their disease status at the time of NGS testing: 1) patients with myeloma but no myelodysplasia (MM_Only, 105 patients and 53.57%); 2) Patients with myelodysplasia but no overt residual myeloma (Myelodysplasia_Only, 14 patients, 7.14%); 3) Patients with both myeloma and myelodysplasia (MM+Myelodysplasia, 27 patients, 13.78%); 4) Patients with neither myeloma or myelodysplasia (Negative_for_Both, 50 patients, 25.51%). The "Myelodysplasia" in this study is defined as having either overt morphologic dysplasia (>10% of the lineage cells), or equivocal dysplasia but having myeloid-related (non-myeloma) cytogenetic abnormalities. NGS results were not included in the classification to assess the added diagnostic value of NGS. The Mutational profiles of the four disease groups are displayed in Figure 1 and compared in Table 1 and 2. The MM+Myelodysplasia group showed highest percentage of mutations (88.89% of patients tested), followed by Myelodysplasia_Only group (57.14%) and MM_Only group (35.24%), with Negative_for_Both group showing the lowest mutation rate (30.00%). The average number of somatic mutations/case also followed the same order: 1.63, 1.00, 0.48, and 0.36, respectively. Of the 196 patients, 58 patients (29.59%) had no morphologic dysplasia or myeloid-related cytogenetic abnormalities but showed one or more somatic mutations by NGS. These patients harbored clonal cytopenia of uncertain significance (CCUS) clones and would have been missed without NGS testing. Of these 58 patients, retrospective review actually identified 7 patients with morphologic dysplasia and were reclassified as MDS. Further mutational analysis revealed the following interesting findings. ASXL1, DNMT3A, KRAS, and SF3B1 mutations showed highest frequencies in MM+Myelodysplaisa group when compared with other 3 groups (Table 2), indicating a close association with myelodysplasia development in patients with persistent myeloma. In contract, among the 4 groups, RUNX1 mutations were most common in Myelodysplasia_only patients, suggesting a potential alternative pathway for myelodysplasia development in patient with myeloma in remission. It is possible that presence of myeloma clones create different evolution pressure on neoplastic myeloid clones. TP53 mutations were present in MM_Only group, but were much more frequent in patients with MM+Myelodysplasia and Myelodysplasia_only groups. The presence of TP53 mutations might therefore suggest increased risk for myelodysplasia. Finally, TET2 were similar between these groups and therefore not of significant diagnostic value. Conclusion: NGS testing is valuable in identifying CCUS, MDS, or tMN in myeloma patients, especially in those with no morphologic or cytogenetic abnormalities. Statistically significant differences are seen in the mutational profiles of the four groups of patients, suggestive of different roles in myelodysplasia development. Further studies are necessary to better distinguish the origin of these mutations as being derived from the myeloma versus the myeloid components of the disease. Disclosures Hussaini: Stemline: Consultancy; Amgen: Consultancy; Janssen: Consultancy; Adaptive: Consultancy; Boston Biomedical: Consultancy. Shain:Karyopharm: Research Funding, Speakers Bureau; AbbVie: Research Funding; Takeda: Honoraria, Speakers Bureau; Sanofi/Genzyme: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Amgen: Speakers Bureau; GlaxoSmithKline: Speakers Bureau; Adaptive: Consultancy, Honoraria; BMS: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau; Janssen: Honoraria, Speakers Bureau; Celgene: Honoraria, Membership on an entity's Board of Directors or advisory committees, Speakers Bureau. Nishihori:Novartis: Other: Research support to institution; Karyopharm: Other: Research support to institution.

Complete reendothelialization followed by inhibition of smooth muscle cell (SMC) proliferation is considered as an effective therapeutic option to prevent restenosis. We have designed poly(lactide-co-glycolide)-loaded bilayered nanoparticles (NPs) with the ability to sequentially release vascular endothelial growth factor (VEGF)-encoding plasmids from the outer layer and paclitaxel (PTX) from the core to promote endothelial regeneration as well as prevent restenosis. Comparing with conventional NPs, which release VEGF plasmid and PTX simultaneously, we expect that the bilayered NPs could release the VEGF plasmid more rapidly, followed by a delayed release of PTX, resulting in an efficient VEGF gene transfection, which ideally could promote reendothelialization and inhibit excessive SMC growth. Indeed, in the present study, we have observed efficient gene transfection using a model plasmid as well as cell growth attenuation in vitro using Chinese hamster ovary cells. Therapeutic efficacy of the bilayered NPs on restenosis was further evaluated in vivo using a rabbit model of atherosclerosis. The bilayered NPs were administered locally via balloon angioplasty to the injured aortic wall through perfusion. Twenty-eight days after the NP administration, rabbits treated with the bilayered NPs exhibited rapid reendothelialization and inhibition of restenosis, as demonstrated by histological analysis. Increased level of VEGF and decreased level of C-reactive protein, a biological marker that is closely related to atherosclerosis, were also observed from animals treated with the bilayered NPs, implicating ameliorated atherosclerosis. Our results suggest that the VEGF plasmid-/PTX-loaded bilayered NPs exert a beneficial impact on atherosclerotic restenosis by sequentially releasing VEGF and PTX in vivo.

Chronic myelomonocytic leukemia (CMML) and juvenile myelomonocytic leukemia (JMML) are myelodysplastic syndrome (MDS)/myeloproliferative neoplasm (MPN) overlap disorders characterized by monocytosis, myelodysplasia, and a characteristic hypersensitivity to granulocyte-macrophage colony-stimulating factor (GM-CSF). Currently, there are no available disease-modifying therapies for CMML, nor are there preclinical models that fully recapitulate the unique features of CMML. Through use of immunocompromised mice with transgenic expression of human GM-CSF, interleukin-3, and stem cell factor in a NOD/SCID-IL2Rγnull background (NSGS mice), we demonstrate remarkable engraftment of CMML and JMML providing the first examples of serially transplantable and genetically accurate models of CMML. Xenotransplantation of CD34+ cells (n = 8 patients) or unfractionated bone marrow (BM) or peripheral blood mononuclear cells (n = 10) resulted in robust engraftment of CMML in BM, spleen, liver, and lung of recipients (n = 82 total mice). Engrafted cells were myeloid-restricted and matched the immunophenotype, morphology, and genetic mutations of the corresponding patient. Similar levels of engraftment were seen upon serial transplantation of human CD34+ cells in secondary NSGS recipients (2/5 patients, 6/11 mice), demonstrating the durability of CMML grafts and functionally validating CD34+ cells as harboring the disease-initiating compartment in vivo. Successful engraftments of JMML primary samples were also achieved in all NSGS recipients (n = 4 patients, n = 12 mice). Engraftment of CMML and JMML resulted in overt phenotypic abnormalities and lethality in recipients, which facilitated evaluation of the JAK2/FLT3 inhibitor pacritinib in vivo. These data reveal that NSGS mice support the development of CMML and JMML disease-initiating and mature leukemic cells in vivo, allowing creation of genetically accurate preclinical models of these disorders.

PurposeClimate change will have a significant impact on China’s potential agricultural production and change the distribution of the population in various regions of China, thus producing population migration. This paper aims to analyze China’s population migration in response to climate change and its socio-economic impact.Design/methodology/approachIn this paper, the Potential Agriculture Production Index is introduced as an analytical tool with which to estimate the scale of the population migration induced by climate change. Also, this paper constructs a multi-regional computable general equilibrium (CGE) model and analyzes the effect of change in the population distribution pattern on regional economies, regional disparity and resident welfare.FindingsThe key finding of this paper is that, as a result of changes in potential agricultural production induced by climate change, the Circum-Bohai-Sea region, the industrialized region and the industrializing region, which are the main destination regions of the migrating population, will face a severe labor shortage. In response to population migration, the economic growth rate of the immigrating population regions has accelerated. Correspondingly, the economic growth rate of the emigrating population regions has decreased. In addition, the larger the scale of population migration is, the larger the economic impact is. Migration increases inner-regional disparity and decreases inter-regional disparity. However, overall regional disparity is only somewhat decreased.Originality/valueThis paper introduces a Potential Agriculture Production Index to estimate the scale of the population migration and introduce a multi-regional CGE model to analyze the correlated social-economic impacts.

BACKGROUND Diabetic nephropathy (DN) is a major microvascular complication of diabetes. Pentraxin 3 (PTX3) is a member of the acute-phase reactants superfamily and altered plasma levels of PTX3 are associated with DN. We performed a case-control study to analyze the relationship between single nucleotide polymorphisms (SNPs) in PTX3 and the risk for DN in patients with type 2 diabetes. MATERIAL AND METHODS The study included 135 DN patients, 155 non-diabetic nephropathy (NDN) patients, and 152 normal controls (NC) (N=442). We genotyped eight PTX3 SNPs (rs2305619, rs2120243, rs1456099, rs7634847, rs1840680, rs2316706, rs2316709, and rs7616177) using the ABI PRISM SNapshot method. RESULTS The genotype frequencies of rs2305619 and rs2120243 differed significantly between the DN and the NDN groups (p=0.017 and p=0.033, respectively). Patients with the GG variant of rs2305619 showed 4.078-fold higher susceptibility to DN than those with the AA variant (OR=4.078, 95% CI=1.370-12.135, p=0.012); patients with the AA variant of rs2120243 had a lower risk of developing DN (OR=0.213, 95% CI=0.055-0.826, p=0.025). Haplotype analysis showed an association between the CAGGG haplotype in block 1 with DN (p=0.0319). CONCLUSIONS Our findings suggested that PTX3 polymorphisms were associated with an increased risk for DN in Chinese patients with type 2 diabetes.

Objective. Patients with diabetic foot ulcers undergoing amputations have poor prognosis. Malnutrition usually occurs in this population and is associated with increased risk of mortality. The geriatric nutritional risk index (GNRI) is a widely used, simple, and well-established tool to assess nutritional risk. The purpose of this study was to assess the association between GNRI and all-cause mortality in diabetic foot ulcers patients undergoing minor or major amputations.Methods. This was a retrospective cohort study including 271 adult patients. Patients were divided into two groups according to a GNRI cutoff value of 92, and characteristics and mortality were compared between the two groups. Cox proportional hazard analysis was performed to explore the association between GNRI and mortality.Result. GNRI (p<0.001), age (p<0.001), and eGFR (p=0.002) were independent predictors of mortality. Among a subgroup of 230 patients with minor amputation, increased age (p<0.001), coronary artery disease (p=0.030), and increased GNRI (p<0.001) were major risk factors.Conclusion. GNRI on admission might be a novel clinical predictor for the incidence of death in patients with diabetic foot ulcers who were undergoing amputations.

An outbreak of severe pseudorabies virus (PRV) infection in farmed mink occurred in northern China in late 2014, causing significant economic losses in the local fur industry. Here, we report the first case of a PRV outbreak in mink in northeastern China, caused by feeding farmed mink with raw pork or organs contaminated by PRV. Mink infected with virulent PRV exhibited diarrhea, neurologic signs, and higher mortality, which can be misdiagnosed as highly pathogenic mink enteritis virus (MEV), canine distemper virus (CDV), and food poisoning. However, these were excluded as causative agents by PCR or bacteria isolation. The duration of disease was 3-7 days, and the mortality rate was 80-90%. PRV was characterized using indirect immunofluorescence assays (IFA) and electron microscopy (EM). Phylogenetic analysis based on full-length genome sequences and those of individual genes of this novel virus strain showed that it clustered in an independent branch with several other PRV isolates from China.

The relative response factors (RRFs) of ten cefazolin impurities were determined by quantitative nuclear magnetic resonance (qNMR) and high-performance liquid chromatography (HPLC) equipped with an ultraviolet (UV) detector. The purities of these ten cefazolin impurities were successfully measured by qNMR for the purpose of RRFs determination by HPLC. The RRF values and their uncertainties determined by the two approaches are comparable. While the qNMR approach is effective and makes it easier to determine the RRFs for impurities, it also has the advantage of allowing the universal detection of protons without the limitations of common mass detectors. The use of qNMR provides a reliable and universal method for the RRF determination of impurities.

Liver cancer is the second leading cause of cancer-related deaths worldwide, and hepatocellular carcinoma is the most common type. The pathogenesis of hepatocellular carcinoma is concealed, its progress is rapid, its prognosis is poor, and the mortality rate is high. Therefore, novel molecular targets for hepatocellular carcinoma early diagnosis and development of targeted therapy are critically needed. Glypican-3, a cell-surface glycoproteins in which heparan sulfate glycosaminoglycan chains are covalently linked to a protein core, is overexpressed in HCC tissues but not in the healthy adult liver. Thus, Glypican-3 is becoming a promising candidate for liver cancer diagnosis and immunotherapy. Up to now, Glypican-3 has been a reliable immunohistochemical marker for hepatocellular carcinoma diagnosis, and soluble Glypican-3 in serum has becoming a promising marker for liquid biopsy. Moreover, various immunotherapies targeting Glypican-3 have been developed, including Glypican-3 vaccines, anti- Glypican-3 immunotoxin and chimeric-antigen-receptor modified cells. In this review, we summarize and analyze the structure and physicochemical properties of Glypican-3 molecules, then review their biological functions and applications in clinical diagnosis, and explore the diagnosis and treatment strategies based on Glypican-3.

Background The differential diagnosis of Parkinson’s disease (PD) with multiple-system atrophy (MSA) is difficult because of their similarity in symptoms and signs. The objective of this study was to investigate the value of external anal-sphincter electromyography (EAS-EMG) and urethral-sphincter electromyography (US-EMG) in differentiating MSA from PD. Methods A total of 201 patients, — 101 MSA and 100 PD — were recruited in this study. Average duration and amplitude of motor unit potentials (MUPs), percentage of polyphasic MUPs, amplitude during strong contractions, and recruitment patterns during maximal voluntary contractions were recorded and analyzed to assess diagnostic efficiency of EAS-EMG and US-EMG for MSA. Results Significant differences in average MUP duration and recruitment patterns during maximal voluntary contractions were found between patients with MSA and patients with PD using both EAS-EMG (P

Polydopamine nanoparticles (PDA NPs) were prepared via dopamine self-polymerization; then, tumor cell lysate (TCL) was covalently attached onto the PDA NPs. The TCL loading capacity was 480 μg per mg of PDA NPs, and the resulting TCL@PDA NPs (241.9 nm) had perfect storage stability and negligible cytotoxicity against APCs. Tumor-bearing mice vaccinated with TCL@PDA NPs experienced significant delay in tumor progression due to the sufficient amount of CTLs and M1-type TAM as well as the deficient number of immunosuppression-related cells in the tumor tissues. Furthermore, empty PDA NPs had the ability to modulate DC maturation and delayed the development of tumors by facilitating the production of activated T cells and decreasing the subpopulation of MDSCs within the tumor microenvironment. Overall, these PDA NPs are expected to be a promising candidate for application as antigen delivery carriers because of their facile antigen loading method as well as their simple and rapid preparation process.

Background: Glycogen Synthase Kinase (GSK)-3β and Brain-derived Neurotrophic Factor (BDNF) play vital roles in both Mild Cognitive Impairment (MCI) and Type 2 Diabetes Mellitus (T2DM). The underlying mechanisms may involve inflammation and oxidative stress. Objectives: To investigate the association of the GSK-3β/BDNF ratio with MCI in elderly patients with T2DM and whether GSK-3β/BDNF ratio can serve as a new diagnostic biomarker for MCI in comorbid with T2DM (MD). Methods: A total of 326 old Chinese T2DM patients were included and stratified according to cognition and GSK-3β/BDNF ratio quartiles. MCI was diagnosed according to the National Institute on Aging Alzheimer’s Association workgroups criteria. In addition to routine hematuria and biochemical examinations, Montreal Cognitive Assessment (MoCA) scale was also used to evaluate the cognitive function, and ELISA method was used to measure GSK-3β activity and the serum levels of BDNF, interleukin 1β (IL-1β), high mobility group box-1 (HMGB1) protein, Malonaldehyde (MDA) and 8-isoprostaglandinF2α (8-iso-PGF2α). Results: We found that GSK-3β activity was negatively correlated with BDNF (r=-0.270, P=0.008), and patients with higher GSK-3β/BDNF ratio had lower MoCA scores (P=0.001). When compared with T2DM patients without MCI (nMD), MD patients had higher GSK-3β activity and GSK-3β/BDNF ratio, but lower BDNF levels. As for inflammation and oxidative stress, IL-1β was inversely correlated with GSK-3β activity, while 8-isoPGF2α was positively correlated with GSK-3β activity and GSK-3β/BDNF ratio. The odds ratio for MCI increased gradually when GSK-3β/BDNF ratio quartile rose from the lowest to the highest (6.90, 95% CI 3.22-14.78). MoCA score was conversely related to GSK-3β/BDNF ratio, age and fast blood glucose (FBG), with GSK-3β/BDNF ratio having the most significant influence on cognition (β=-0.199, P Conclusion: Our data provide evidence for a strong link between GSK-3β/BDNF ratio and MCI. GSK- 3β/BDNF ratio may serve as a better diagnostic biomarker for MD than either GSK-3β or BDNF alone and increased GSK-3β/BDNF ratio indicates a worse cognitive function.

The economic damage function of the integrated assessment model (IAM) is used to quantify the economic loss caused by climate change, and it is quite important for coupling the economic system and natural system in a model. However, there are many shortcomings of the damage function in the current IAM. The most important shortcoming is that the only climatic factor in the damage function is the global annual mean temperature rise, which would result in the estimation deviating from reality. We improved the damage function by introducing extreme climate change events. Additionally, the improved damage function is applicable to the regional characteristics by using regional climate change data. The extreme climate event information is extracted with the categories of the disasters. The damage function is set up by using the Ordinary Least Square (OLS) method based on the climate data and economic loss data. The results show that the improved IAM damage function can better reflect the actual climate change economic loss in China, making it more reasonable. This paper provides an important method and technical solution for the realization of an effective connection between the disaster damage function of the integrated assessment model and the climate elements of the Earth system model.

Corynebacterium glutamicum is the main industrial strain to produce L-valine by microbial fermentation. In this study, a low L-alanine producing C. glutamicum strain VWB-2 was constructed by knocking out the alanine aminotransferase encoding gene alaT in a high L-valine producing strain VWB-1. Meanwhile, a site-directed mutagenesis (ilvBN₁ (M13)) was done on the regulatory subunit of acetohydroxyacid synthase (ilvBN), a key enzyme in the L-valine synthesis pathway. Furthermore, the overexpression of the genes involved in the biosynthesis of L-valine, the mutated ilvBN₁ (M13), the acetohydroxy acid isomerase coding genes ilvC, the dihydroxy-acid dehydratase coding gene ilvD and branched-chain amino acid aminotransferase coding gene ilvE, could all promote the L-valine production of VWB-1 by strengthening the carbon flow towards L-valine. With the overexpression of the branched chain amino acid transporter coding gene brnFE and its regulator lrp₁, the L-valine producing capability of VWB-1 was further enhanced. The finally obtained engineered strain VWB-2/pEC-XK99E-ilvBN₁ (M13)CE-lrp₁-brnFE could produce 461.4 mmol/L L-valine in a 5 L fermentor with a sugar acid conversion rate of 0.312 g/g glucose.谷氨酸棒状杆菌是目前微生物发酵生产L-缬氨酸的主要工业菌株。文中首先在谷氨酸棒状杆菌 VWB-1 中敲除了alaT (丙氨酸氨基转移酶)，获得突变菌株VWB-2，作为出发菌株。进而对L-缬氨酸合成途径关键酶——乙酰羟酸合酶 (ilvBN) 的调节亚基进行定点突变 (ilvBN₁ (M13))，解除L-缬氨酸对该酶的反馈抑制。然后辅助过量表达L-缬氨酸合成途径关键基因ilvBN₁ (M13)、乙酰羟酸异构酶 (ilvC)、二羟酸脱水酶 (ilvD)、支链氨基酸氨基转移酶 (ilvE)，加强通往L-缬氨酸的碳代谢流，提高菌株的L-缬氨酸水平。最后，基于过量表达L-缬氨酸转运蛋白编码基因brnFE 及其调控蛋白编码基因lrp1，提高细胞的L-缬氨酸转运能力。最终获得工程菌株VWB-2/pEC-XK99E-ilvBN₁ (M13)CE-lrp₁-brnFE 在5 L 发酵罐中的L-缬氨酸产量达到461.4 mmol/L，糖酸转化率达到0.312 g/g 葡萄糖。.

Mink enteritis virus (MEV) is a major pathogen inducing acute hemorrhagic enteritis in mink. This study aims to determine the pathogenicity of the isolated MEV strain (SMPV-11) compared with the attenuated MEV strain (MEV-F61) in the mink. The two MEV strains were inoculated in the two mink groups, respectively. Then the clinical symptom, hematological, serological, and histopathological change were evaluated. Our findings showed that there were differences in the clinical features and pathological changes of the SMPV-11 and MEV-F61 in the mink. It indicates that SMPV-11 is a virulent strain, and it can be the potential MEV vaccine strain in the mink.

In chalcocite (Cu2S) bioleaching, the lack of iron metabolism is a key restricting factor. As the most common sulfide mineral, pyrite (FeS2) can release Fe(Ⅱ) and compensate for the iron metabolism deficiency in chalcocite bioleaching. The bioleaching of chalcocite in an imitated industrial system was improved by enhancing the iron–sulfur metabolism simultaneously using pyrite and sulfur oxidizers based on the joint utilization of waste resources, while the bioleaching performance and community structure in the leachate were systematically investigated. Due to the active sulfur/iron metabolism, the pH reached 1.2, and Fe3+ was increased by 77.78%, while the biomass of planktonic cells was improved to 2.19 × 107 cells/mL. Fourier transform infrared reflection (FTIR) and X-ray diffraction (XRD) analysis results showed that more iron–sulfur crystals were produced due to more active iron–sulfur metabolism. Scanning electron microscopy (SEM) revealed that many derivative particles and corrosion marks appeared on the surface of the ore, implying that the mineral–microbe interaction was strengthened. Confocal laser scanning microscopy (CLSM) showed the accumulation of cells and extracellular polymeric substances (EPS) on the ore surface, indicating a stronger contact leaching mechanism. Furthermore, the community structure and canonical correspondence analysis (CCA) demonstrated that the introduction of sulfur-oxidizing bacteria and pyrite could maintain the diversity of dominant leaching microorganisms at a high level. Sulfobacillus (27.75%) and Leptospirllillum (20.26%) were the dominant sulfur-oxidizing and iron-oxidizing bacteria during the bioleaching process. With the accumulation of multiple positive effects, the copper ion leaching rate was improved by 44.8%. In general, this new type of multiple intervention strategy can provide an important guide for the bioleaching of low-grade ores.

Bioaugmentation was conducted using a bacterial consortium of Pseudomonas putida SW-3 and Rhodococcus ruber SS-4, to test their ability to degrade benzene, toluene, and styrene (BTS). SW-3 and SS-4 were isolated from domestic sludge and sewage samples to establish a synthetic consortium with an optimized ratio of 2:1 to reach a degradation efficiency of 82.5–89.8% of BTS. The bacterial consortium was inoculated with sludge and sewage samples at a ratio of 2:1, resulting in a degradation efficiency of 97.9% and 92.7%, respectively, at a BTS concentration of 1800 mg·L−1. Analysis of bacterial community structure following bioaugmentation indicated an increase in abundance of BTS-degrading bacteria, particularly Acinetobacter and Pseudoxanthomonas in sludge and Pseudomonas in sewage, enhancing the collective BTS degradation ability of the bacterial community. Principal component analysis demonstrated that a more balanced bacterial community structure was established following intervention. This indicated that the selected bacteria are excellent candidates for bioaugmentation.

Acute myeloid leukemia (AML) with CBFB-MYH11 fusion gene is almost always associated with either inv(16)(p13.1q22) or t(16;16)(p13.1;q22). Only a few variants leading to formation of this chimeric gene have been reported. In this report, we describe a novel type of CBFB-MYH11 fusion variant generated by an insertion of CBFB into the MYH11 gene, which is identifiable by fluorescence in situ hybridization (FISH).

Bioleaching microorganisms inhabit extremely acidic environments (pH < 3.0) and can be used in the biohydrometallurgical industry. They employ several strategies, such as biofilms formation, mechanical defense, membrane reversal potential, proton efflux, intracellular buffering, and protection, or repair mechanism of biomacromolecules, to maintain their intracellular pH within a narrow range, which is close to neutral, for conducting normal physiological activity. In this review, we describe the effects of these strategies on the homeostasis of intracellular pH of bioleaching microorganisms and the relevant energy metabolism. The potential significance of horizontal gene transfer and gene loss in their adaptation to the environment, and the prospect of new technologies, such as cryo EM technology, in revealing the potential acid-resistant components have also been discussed.

The sustainable development of agriculture is important for China, where more than 20 percent of the world population live. Many factors affect the sustainable development of agriculture in China. The purpose of this paper is to find out what factors play influential roles on affecting the growth pattern. Based on a modified production function model built upon Slow (1956) and Romer (2001), the impact of growth drag on the agricultural economy, and the contribution resources, are evaluated empirically using a time-series regression analysis based on the case of China for the period 1978&ndash, 2015. The estimated coefficient of the total agricultural growth drag in China is 1.32, which suggests that the annual agricultural growth is associated with a 1.32 percent decline due to the scarcities of land, water, and energy. Specifically, the growth drags from land, water, and energy are found to be 0.5 percent, 0.44 percent, and 0.38 percent, respectively. Additionally, the results suggest that capital is the most important factor in the agricultural sector, although agriculture is highly dependent on land. Capital contributes around 76.86 percent to the growth of the agricultural economy, whereas contributions from land, energy, water, and technological progress are relatively small. Therefore, more investment should be added to agriculture, to accelerate the technology progress. Furthermore, the saving of water and energy is also important for the sustainable development of agriculture.

Introduction: Advanced molecular profiling with next generation sequencing (NGS) has greatly improved the diagnostic and prognostic stratification of hematological disorders. Several driver mutations including TP53, RUNX1, ASXL1, and EZH2 mutations are independent predictors of overall survival (OS) in myelodysplastic syndromes (MDS). Frequent co-mutations of ASXL1, EZH2,RUNX1 and/or TET2 have also been described in the entity. However, studies exploring the clinical significance of co-mutations in these adverse genes are limited. Our study assesses mutation profiles and compares clinical outcomes among MDS patients with EZH2 mutation alone, ASXL1 mutation alone, ASXL1/EZH2 co-mutation, and co-mutated ASXL1/EZH2 with additional mutations. Materials and Methods: The retrospective study was approved by the institutional review board. We searched our institutional database (>4000 patients) for patients who were diagnosed with MDS and harbored only ASXL1 or EZH2 mutations, only ASXL1/EZH2 co-mutations, and co-mutated ASXL1/EZH2 plus other genes identified by targeted NGS using a myeloid gene panel (up to 54 genes). Patient demographics, diagnosis, cytogenetic abnormalities, and overall survival times were analyzed. Mann-Whitney U Test was used for statistical analysis of OS and a Kaplan Meier survival curve was generated. Results: Total 140 patients were retrieved: 71 patients with only ASXL1 mutation, 12 patients with only EZH2 mutation, 8 patients with only ASXL1/EZH2 co-mutation and 49 patients with multiple mutations. As shown in Figure 1, patients with ASXL1/EZH2 co-mutation or multiple mutations show worse OS when compared with patients with EZH2 mutation alone or ASXL1 mutation alone. However, only the differences between either the survival of patients with ASXL1 or EZH2 mutations alone vs multiple mutations are approaching statistical significance (p values = 0.0669 and 0.0927 respectively). Interestingly the patients with ASXL1 mutation alone or multiple mutations appeared more likely to harbor good cytogenetics (70.42% and 65.3%) rather than poor cytogenetics (7% and 2%, respectively). Finally, TP53 mutation appears to be mutually exclusive with ASXL1 or EZH2 mutations, occurring only in 1 patient with multiple mutations. Conclusion: In our study, MDS patients with ASXL1/EZH2 co-mutations or combined with other mutations appear to show worse prognosis than patients harboring ASXL1 mutation alone or EZH2 mutation alone, though statistical significance was not reached. This could be due to the fact that both ASXL1 and EZH2 confer such an unfavorable prognosis to the patients that additional mutation(s) will not produce any significant clinical impacts. This could also be due to the limited sample size in our study, thus the limited statistical power. A larger cohort study is needed to further explore the prognostic values of co-mutations in this setting. Similar to other reports, TP53 mutation also appears to be mutually exclusive with ASXL1 and/or EZH2; that is of value to further explore the molecular mechanisms. Figure 1 Disclosures Sallman: Abbvie: Speakers Bureau; Novartis: Speakers Bureau; Jazz: Research Funding; Incyte: Speakers Bureau; Celyad: Membership on an entity's Board of Directors or advisory committees; Celgene: Research Funding, Speakers Bureau. Komrokji:celgene: Consultancy; pfizer: Consultancy; DSI: Consultancy; Novartis: Speakers Bureau; JAZZ: Speakers Bureau; Incyte: Consultancy; Agios: Consultancy; JAZZ: Consultancy.

Improvements in antisense technology have now enabled clinically relevant therapeutic credentialing of the noncoding genome. MALAT1 is a long non-coding RNA that, among other functions, is thought to serve as a nuclear scaffold for splicing and transcription factors. MALAT1 expression is associated with inferior prognosis across solid tumors and its depletion impairs proliferation and metastasis in preclinical solid tumor models. We found that elevated MALAT1 levels are independently associated with inferior overall survival in patients with CMML. Further, RNA-sequencing of primary CMML monocytes identified MALAT1 as the fourth most over-expressed transcript compared to controls. Therefore, we explored the biologic relevance and therapeutic candidacy of MALAT1 across several human and murine models of CMML. First, we crossed NRASQ61R/+Mx1-cre driven mice, which display a CMML-like phenotype, to MALAT1KO/KOmice. Although MALAT1KO/KOmice did not have abnormalities in complete blood counts, immunophenotyping of the hematopoietic stem cell (HSC) compartment identified statistically significantly lower numbers of HSC compared to wild type (WT) controls and a non-significant decrease in NRASQ61R/+/MALAT1KO/KOcompared to NRASQ61R/+alone. This decrease in HSC was not a result of impaired self-renewal as no differences were observed in these models after in vivo competitive transplant experiments. Therefore, we reasoned that MALAT1 expression may be controlling HSC differentiation. To test this, we transformed bone marrow cells from these models with an estrogen-regulated (ER) Hoxb8 construct enabling cells to maintain an HSC state until ER is withdrawn and myeloid differentiation is induced. ER-Hoxb8NRASQ61R/+/MALAT1KO/KOcells had increased basal levels of Gr-1 compared to ER-Hoxb8 transformed NRASQ61R/+alone that was dramatically enhanced upon ER withdrawal suggesting that MALAT1 depletion regulates myeloid differentiation. These findings were validated by assessment of morphology, transcriptome, and in vivo immunophenotyping of bone marrow and spleen cells. Further, moribund NRASQ61R/+/MALAT1KO/KOmice displayed a reduction in organomegaly typically associated with leukemic burden. We validated this in human monocytic leukemia by generating MALAT1 depleted THP-1 isogeneic cell lines using the CRISPR/Cas9 system. MALAT1 depleted THP-1 cells (MKO) had greater terminal differentiation according to immunophenotypic markers and morphology that was greatly enhanced when treated with phorbol myristate acetate. Last, MKO orthotopic xenografts demonstrated inferior human leukemia engraftment and decreased spleen and liver weights, and heterotopic xenografts exhibited reduced tumor volume, collectively suggesting diminished leukemic burden. Because ATRA has been clinically tested in CMML with minimal effects, we next explored whether MALAT1 depletion could potentiate ATRA differentiation in CMML. First, we treated MKO cells with ATRA and observed a large induction of myeloid differentiation by marker expression and morphologic assessment compared to isogenic controls. This was validated by NRASQ61R/+/MALAT1KO/KOmice demonstrating that ATRA more robustly induced differentiation compared to vehicle which was not seen in NRASQ61R/+/MALAT1+/+mice. Next, we tested MALAT1 antisense oligonucleotides (ASOs) currently under clinical development in THP-1 cells +/- ATRA and demonstrated both an increase in myeloid differentiation and apoptosis compared to ATRA alone. To test this therapeutic strategy in primary CMML specimens, we generated CMML patient-derived xenografts (n=30 mice) and treated each with ASO, ATRA, the combination, or controls and identified a more robust reduction in human HSC engraftment with the combination. To explore the mechanistic basis for these findings, we performed RNA-sequencing of MALAT1-depleted or control cells and identified that CREB target genes were differentially expressed. Basal protein levels of p-CREB were also decreased in MKO cells and were further reduced in the nucleus of MKO by western and microscopy. Lastly, overexpression of WT or constitutively active CREB but not its dominant negative rescued the differentiation effect seen in ATRA treated MKO cells. Taken together, MALAT1 is a novel, CREB-dependent regulator of myeloid differentiation and its depletion potentiates ATRA therapy. Disclosures Cluzeau: Menarini: Consultancy; Jazz Pharma: Consultancy; Abbvie: Consultancy. Komrokji:celgene: Consultancy; pfizer: Consultancy; DSI: Consultancy; JAZZ: Speakers Bureau; Novartis: Speakers Bureau; JAZZ: Consultancy; Agios: Consultancy; Incyte: Consultancy. MacLeod:Ionis Pharmaceuticals: Employment. List:Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding. Epling-Burnette:Forma Therapeutics: Research Funding; Celgene Corporation: Patents & Royalties, Research Funding; Incyte Corporation: Research Funding.

Background: DNMT3A and SF3B1 are two of the most commonly mutated genes in myeloid neoplasms. DNMT3A mutations have been found to be associated poor prognosis in acute myeloid leukemia (AML), while SF3B1 mutations have been reported to be associated with favorable prognosis in myelodysplastic syndrome (MDS). Studies exploring the clinical significance of DNMT3A and SF3B1 co-mutation are limited and conflicting. Furthermore, the previous studies did not control for the presence of other co-mutated genes, which could be confounding factors. Materials and Methods: 4390 in-house patients with confirmed or suspected myeloid neoplasms were tested by Next Generation Sequencing (NGS) (up to 54 gene panel). From these, we identified 68 patients with only DNMT3A mutation, 61 patients with only SF3B1 mutation, and 23 patients with only SF3B1/DNMT3A co-mutation, for a total of 152 patients. Patient demographics, diagnosis, cytogenetic abnormalities, and overall survival time were collated and analyzed. Results: Patients with SF3B1 mutation alone or SF3B1/DNMT3A double mutations were found to be significantly older (median age of 76 and 75, respectively) than patients with isolated DNMT3A mutation (median age of 66, p values = 0.00016 and 0.00174 respectively). Patients with SF3B1/DNMT3A double mutations or isolated SF3B1 mutations occurred more commonly in MDS. They occurred less frequently in acute myeloid leukemia (AML) (1.63% and 21.74% respectively) than isolated DNMT3A mutation (41.76%, p values < 0.00001 and p = 0.00529 respectively). Patients with double mutations demonstrated increased ring sideroblasts (100%) at a rate similar to patients with isolated SF3B1 mutation (92.16%), which are unsurprisingly higher than patents with DNMT3A mutation alone (8%, p value < 0.000001). The patients with SF3B1 mutation alone or SF3B1/DNMT3A co-mutation were less likely to have poor cytogenetics and more likely to have intermediate cytogenetics than patients with DNMT3A mutation alone. Finally, patients with sole SF3B1 mutation or SF3B1/DNMT3A co-mutation showed significantly longer median survival time (26.60 months and 19.93 months respectively) and better overall survival curve (Figure 1) than patients with DNMT3A mutation alone (8.32 months, p value = 0.001 and 0.01078 respectively), for all the patients in this study as well as for patients with only myelodysplastic syndrome. Conclusion: Patients with SF3B1/DNMT3A double mutations showed a prognosis similar to patients with isolated SF3B1 mutation and better clinical outcomes than patients with isolated DNMT3A mutation. This suggests that the favorable prognosis of SF3B1 mutation in MDS or AML patients is not abrogated by the concurrent presence of a DNMT3A mutation. Figure 1 Disclosures No relevant conflicts of interest to declare.

To investigate the effect of hydrogen-rich water on cerebral edema and aquaporin 1 (AQP1) expression in rats with traumatic brain injury (TBI).Ninety male Sprague-Dawley (SD) rats were randomly divided into sham operation group, TBI model group, hydrogen-rich water treatment group (H group),with 30 rats in each group. TBI model was reproduced by weight dropping method. The skulls of rats in sham operation group underwent only craniotomy without direct hit and with bone wax sealed suture.5 mL/kg of hydrogen-rich water injection was given intraperitoneally after model reproduction in H group, and equal amount of normal saline was given in sham and TBI groups, once a day for both groups for 5 days. Six rats from each group were sacrificed at 6,12,24,48 hours and 5 days after evaluating neurological severity scores (NSS).The cerebral cortex was harvested, and the pathological changes in morphology of brain tissue were observed with light microscope. The positive expression of AQP1 in cerebral cortex was observed with immunohistochemistry by light microscopy, the AQP1 mRNA expression in cerebral cortex was determined by real-time fluorescent quantization reverse transcription-polymerase chain reaction (RT-PCR),and the AQP1 protein expression in cerebral cortex was determined by Western Blot.① All rats in sham operation group had a NSS of zero at each time point. NSS of TBI group was obviously raised with time prolongation, and peaked at 24 hours followed by a lower tendency, while the score in H group was significantly lower than that of TBI group, and the difference was the most obvious at 24 hours as compared with TBI group (9.83 ± 2.78 vs.13.50± 2.42,P ＜ 0.05).② It was shown by light microscope that in the TBI group there were pathological changes in cerebral cortex, including obvious irregular arrangement of nerve cells, cerebral edema, obvious bleeding, especially at 24 hours, then the cerebral edema became vanished gradually; and the positive expression of AQP1 in the pia mater at all the time points in the TBI group was significantly increased, and it was most obvious at 24 hours. Compared with TBI group, the pathological changes at time points of 12 hours to 5 days in H group was significantly lessened, and the positive expression of AQP1 in the cerebral pia mater was reduced obviously.③ Compared with sham operation group, the mRNA and protein expressions of AQP1 in cerebral cortex in TBI group were significantly elevated, peaked at 24 hours [AQP1 mRNA (2-△△Ct):7.50±0.26 vs.1,AQP1 protein (gray value):1.986±0.110 vs.0.336±0.034, both P ＜ 0.05], then they gradually declined. The mRNA and protein expressions of AQP1 in cerebral cortex were significantly decreased after hydrogen-rich water treatment [24-hour AQP1 mRNA (2-△△Ct):5.40±0.21 vs.7.50±0.26, 24-hour AQP1 protein (gray value): 1.246±0.137 vs.1.986±0.110, both P ＜ 0.05].The up-regulation of AQP1 mRNA and protein in rats' cerebral cortex after TBI perhaps participates in edema formation which might be involved in the pathophysiology of cerebral edema in TBI. Early treatment with an intraperitoneally injection of hydrogen-rich water is capable of attenuating the extent of TBI-induced up-regulation of AQP1 mRNA and protein, alleviating cerebral edema, and achieving its protective effects.

Getah virus (GETV), a mosquito-borne virus that mainly infects horses and pigs, has emerged and spread in China. We developed a highly specific and reproducible TaqMan probe-based quantitative reverse transcription PCR (RT-qPCR) assay targeting the non-structural protein 1 of GETV, whose detection limit is 25.5 copies/µL, which is 100-fold higher than that of conventional RT-PCR. RT-qPCR was used to detect GETV RNA in mosquito and animal clinical samples, showing that the accuracy of RT-qPCR was higher than that of conventional RT-PCR. The newly developed RT-qPCR assay may be a useful alternative tool for rapid, simple and specific diagnosis of GETV infection.

Toward the elucidation of the advanced mechanism of l-valine production by Corynebacterium glutamicum, a highly developed industrial strain VWB-1 was analyzed, employing the combination of transcriptomics and proteomics methods. The transcriptional level of 1155 genes and expression abundance of 96 proteins were changed significantly by the transcriptome and proteome comparison of VWB-1 and ATCC 13869. It was indicated that the key genes involved in the biosynthesis of l-valine, ilvBN, ilvC, ilvD, ilvE were up-regulated in VWB-1, which together made prominent contributions in improving the carbon flow towards l-valine. The l-leucine and l-isoleucine synthesis ability were weakened according to the down-regulation of leuB and ilvA. The up-regulation of the branched chain amino acid transporter genes brnFE promoted the l-valine secretion capability of VWB-1. The NADPH and ATP generation ability of VWB-1 were strengthened through the up-regulation of the genes involved in phosphate pentose pathway and TCA pathway. Pyruvate accumulation was achieved through the weakening of the l-lactate, acetate and l-alanine pathways. The up-regulation of the genes coding for elongation factors and ribosomal proteins were beneficial for l-valine synthesis in C. glutamicum. All information acquired were useful for the genome breeding of better industrial l-valine producing strains.

Hemorrhagic pneumonia in mink is a fatal disease caused by Pseudomonas aeruginosa. Very little is known about P. aeruginosa in relation to genotype and the mechanisms underlying antimicrobial resistance in mink. A total of 110 P. aeruginosa samples were collected from mink from Chinese mink farms between 2007 and 2015. Samples underwent molecular genotyping using pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST), antimicrobial susceptibility and its mechanism were investigated at the molecular level. The PFGE identified 73 unique types and 15 clusters, while MLST identified 43 (7 new) sequence types (ST) and 12 sequence type clonal complexes (STCC). Sequence types and PFGE showed persistence of endemic clones in cities Wendeng (Shandong, China) and Dalian (Liaoning, China), even in different timelines. The MLST also revealed the gene correlation of the mink P. aeruginosa across different time and place. The ST1058 (n = 14), ST882 (n = 11), and ST2442 (n = 10) were the predominant types, among which ST1058 was the only one found both in Shandong province and Dalian (Liaoning, China). The MLST for P. aeruginosa infection in mink was highly associated with that in humans and other animals, implying possible transmission events. A small proportion of mink exhibited drug resistance to P. aeruginosa (9/69, 13%) with resistance predominantly to fluoroquinolone, aminoglycoside, and β-lactamase. Eight strains had mutations in the quinolone-resistance determining regions (QRDR). High proportions (65%; 72/110) of the fosA gene and 2 types of glpt deletion for fosmycin were detected. Furthermore, in the whole genome sequence of one multidrug resistant strain, we identified 27 genes that conferred resistance to 14 types of drugs.

Auxin response factors (ARFs) have been reported to play vital roles during plant growth and development. In order to reveal specific functions related to vegetative organs in grasses, an in-depth study of the ARF gene family was carried out in switchgrass (Panicum virgatum L.), a warm-season C4 perennial grass that is mostly used as bioenergy and animal feedstock. A total of 47 putative ARF genes (PvARFs) were identified in the switchgrass genome (2n = 4x = 36), 42 of which were anchored to the seven pairs of chromosomes and found to be unevenly distributed. Sixteen PvARFs were predicted to be potential targets of small RNAs (microRNA160 and 167). Phylogenetically speaking, PvARFs were divided into seven distinct subgroups based on the phylogeny, exon/intron arrangement, and conserved motif distribution. Moreover, 15 pairs of PvARFs have different temporal-spatial expression profiles in vegetative organs (2nd, 3rd, and 4th internode and leaves), which implies that different PvARFs have specific functions in switchgrass growth and development. In addition, at least 14 pairs of PvARFs respond to naphthylacetic acid (NAA) treatment, which might be helpful for us to study on auxin response in switchgrass. The comprehensive analysis, described here, will facilitate the future functional analysis of ARF genes in grasses.