Systemic immune profiling of Omicron-infected subjects inoculated with different doses of inactivated virus vaccine.

SARS-CoV-2 primary strain-based vaccination exerts a protective effect against Omicron variants-initiated infection, symptom occurrence, and disease severity in a booster-dependent manner. Yet, the underlying mechanisms remain unclear. During the 2022 Omicron outbreak in Shanghai, we enrolled 122 infected adults and 50 uninfected controls who had been unvaccinated or vaccinated with two or three doses of COVID-19 inactive vaccines and performed integrative analysis of 41-plex CyTOF, RNA-seq, and Olink on their peripheral blood samples. The frequencies of HLA-DRhi classical monocytes, non-classical monocytes, and Th1-like Tem tended to increase, whereas the frequency of Treg was reduced by booster vaccine, and they influenced symptom occurrence in a vaccine dose-dependent manner. Intercorrelation and mechanistic analysis suggested that the booster vaccination induced monocytic training, which would prime monocytic activation and maturation rather than differentiating into myeloid-derived suppressive cells upon Omicron infections. Overall, our study provides insights into how booster vaccination elaborates protective immunity across SARS-CoV-2 variants.

Sohlh2 Promotes the Progression of Hepatocellular Carcinoma via TGM2-Mediated Autophagy.

Liver cancer is the third leading cause of cancer-related deaths worldwide, with hepatocellular carcinoma (HCC) accounting for 85% of liver cancer-related deaths. Autophagy controls HCC cell growth, invasion, metastasis, drug resistance, and stemness. Spermatogenesis and oogenesis basic helix-loop-helix transcription factor 2 (Sohlh2) can bind to the E-boxes in the promoter regions of target genes, which are involved in multiple neoplasms. In this study, Sohlh2 was highly expressed in HCC tissues and was related to poor prognosis. Moreover, Sohlh2 overexpression promoted the proliferation, migration, invasion, and metastasis of HCC cells in vivo and in vitro. However, Sohlh2 silencing inhibited proliferation, migration, invasion, and metastasis of HCC cells in vivo and in vitro. Mechanistically, Sohlh2 could bind to the promoter of TGM2 and enhance its transcriptional activity, thereby enhancing the autophagy of HCC cells. Furthermore, Sohlh2 protein levels were positively associated with TGM2 expression in HCC tissues. Taken together, these results demonstrate that Sohlh2 can promote HCC progression via TGM2-mediated autophagy, implying that Sohlh2 is a promising candidate for HCC treatment.

Determining the prevalence of Escherichia coli, Salmonella, and shiga toxin-producing Escherichia coli in manure of dairy lagoons.

AIM: The aim of this study was to determine the prevalence of microbial pathogens in manure of dairy lagoons in California. METHODS AND RESULTS: To determine pathogens in dairy manure stored in anaerobic lagoons of dairy farm, an extensive field study was conducted across California to sample manure from 20 dairy farms. Samples were analyzed to determine the prevalence of indicator Escherichia coli, Shiga toxin producing E. coli (STEC), Salmonella, and E. coli O157: H7. To test the E. coli, STEC, and Salmonella, we used agar culture-based method followed by polymerase chain reaction (PCR) method. In addition, a real- time PCR based method was used to determine the presence of E coli O157: H7. Study demonstrated that the prevalence of Salmonella in manure sample is lower than E. coli. The presence of Salmonella was found in 2.26% of the samples, and both the culture-based and PCR methods yielded comparable outcomes in detecting Salmonella. Moreover, ∼11.30% of the total samples out of the 177 were identified as positive for STEC by qPCR. CONCLUSION: These findings demonstrate that indicator E. coli are abundantly present in anaerobic lagoons. However, the presence of STEC, and Salmonella is substantially low.

Down-regulated Wnt7a and GPR124 in early-onset preeclampsia placentas reduce invasion and migration of trophoblast cells.

OBJECTIVES: Preeclampsia (PE) is a disease specific to pregnancy that causes 9-10 % of maternal deaths. Early-onset PE (<34 weeks' gestation) is the most dangerous category of PE. Wnt7a and GPR124 (G protein-coupled receptor 124) are widely expressed in the human reproductive process. Especially during embryogenesis and tumorigenesis, Wnt7a plays a crucial role. However, few studies have examined the association between Wnt7a-GPR124 and early-onset PE. The aim of this study was to examine the significance of Wnt7a and GPR124 in early-onset PE as well as Wnt7a's role in trophoblast cells. METHODS: Immunohistochemistry (IHC), real-time PCR, and western blotting (WB) were used to investigate Wnt7a and GPR124 expression in normal and early-onset PE placentas. Additionally, FACS, Transwell, and CCK-8 assays were used to diagnose Wnt7a involvement in migration, invasion, and proliferation. RESULTS: In the early-onset PE group, Wnt7a and GPR124 expression was significantly lower than in the normal group, especially in the area of syncytiotrophoblasts (STBs) and extravillous trophoblasts (EVTs). A negative correlation was found between Wnt7a RNA and GPR124 expression (r=-0.42, p<0.01). However, the Wnt7a RNA expression level was positive correlated with PE severity. In further cellular functional experiments, knockdown of Wnt7a inhibits HTR8/SVeno cells invasion and migration but has little effect on proliferation and apoptosis. CONCLUSIONS: Through the Wnt pathway, Wnt7a regulates trophoblast cell invasion and migration, and may contribute to early-onset preeclampsia pathogenesis. A molecular level study of Wnt7a will be needed to find downstream proteins and mechanisms of interaction.

Economic and environmental performance of microalgal energy products - A case study exploring circular bioeconomy principles applied to recycled anaerobic digester waste flows.

This study quantifies the financial and environmental impacts of a microalgal bioenergy system that attempts to maximize circular flows by recovering and reusing the carbon, nutrients, and water within the system. The system produces microalgal biomass using liquid digestate of an anaerobic digester that processes 45 metric tons of food waste and generates 28.6 m3 of permeate daily in California, and three energy production scenarios from the biomass are considered: producing biodiesel, electricity, and both. In all scenarios, the resulting energy products delivered only modest reductions in environmental impacts as measured by carbon dioxide equivalent emissions. The carbon intensities (CIs) of biodiesel from this study were 91.0 gCO2e/MJ and 93.3 gCO2e/MJ, which were lower than 94.71 gCO2e/MJ of conventional petroleum diesel, and the CI of electricity from this study was 70.6 gCO2e/MJ, lower than the average electricity grid CI in California (82.92 gCO2e/MJ). The economic analysis results show that generating electricity alone can be profitable, while biodiesel produced via this system is not cost competitive with conventional diesel due to high capital expenses. Thus, generating electricity in lieu of biodiesel appears to be a better option to maximize the use of waste flows and supply lower-carbon energy.

Sharp angular and unidirectional filter based on accidental degeneracy between two twisted Weyl semimetal-based defect modes in a one-dimensional photonic crystal.

To address the challenges associated with the realization of optical non-reciprocity and enhance the efficiency of GaAs solar cells, among other systems, in this study, we investigated defect-mode interactions in a one-dimensional photonic crystal containing two Weyl semimetal-based defect layers. Moreover, two non-reciprocal defect modes were observed, namely, when defects are identical and nearby. Increasing the defect distance weakened the defect-mode interactions, thus causing the modes to gradually move closer and then degenerate into one mode. It should be noted that by changing the optical thickness of one of the defect layers, the mode was found to degrade to two non-reciprocal dots with different frequencies and angles. This phenomenon can be attributed to an accidental degeneracy of two defect modes with dispersion curves that intersect in the forward and backward directions, respectively. Moreover, by twisting Weyl semimetal layers, the accidental degeneracy occurred only in the backward direction, thus resulting in a sharp angular and unidirectional filter.

Identification of eIF6 as a prognostic factor that drives tumor progression and predicts arsenic trioxide efficacy in lung adenocarcinoma.

BACKGROUND: Lung cancer is the leading cause of cancer-related mortality worldwide. Dysregulation of mRNA translation can contribute to the development and progression of cancer whilst also having an impact on the prognosis of different types of malignancies. Eukaryotic translation initiation factors (eIFs) have been reported to serve a key role in the initiation of mRNA translation. However, little was known about the association between eIF6 and lung adenocarcinoma (LUAD) progression. We aimed to elucidate the roles of eIF6 in LUAD tumorigenesis. METHODS: Bioinformatic analysis was conducted to assess the clinical significance of eIF6 in LUAD. CCK-8, colony formation assays were used to evaluate the biological roles of eIF6. The subcutaneous model was used to assess the in vivo roles of eIF6. RESULTS: In the present study, it was found that eIF6 expression was significantly higher in LUAD samples compared with that in normal lung tissues. Higher expression levels of eIF6 were found to be associated with more advanced clinical stages of LUAD and poorer prognoses in patients with LUAD. Subsequently, overexpression of eIF6 was demonstrated to promote LUAD cell proliferation, migration and invasion, which are features of metastasis, in vitro. By contrast, inhibition of eIF6 induced cell cycle arrest and apoptosis in LUAD cells. Further bioinformatics analysis and experimental assays revealed that eIF6 expression positively correlated with the mRNA expression of stemness-associated genes in LUAD cells. Targeting eIF6 suppressed the sphere formation capacity of LUAD cells. In addition, data from the subcutaneous xenograft model in vivo also suggested that eIF6 deficiency could significantly delay tumor growth and improve the prognosis of mice. Targeting eIF6 rendered LUAD cells sensitive to arsenic trioxide treatment. CONCLUSION: The present study suggest that eIF6 can serve as a prognostic biomarker and a potential therapeutic target for patients with LUAD.

Nematode-Suppressive Potential of Digestates to Meloidogyne incognita and Heterodera schachtii.

Management of plant-parasitic nematodes uses host plant resistance, crop rotation, cultural methods, and nematicide applications. Host plant resistance is tedious to develop, and crop rotation and cultural methods are challenging to use. Environmental and human health concerns render sole reliance on chemical nematode suppression nonsustainable. Previously, digestate from anaerobically fermented maize silage suppressed Heterodera schachtii in Beta vulgaris crops. Here, seven digestates were investigated for nematode suppressive potential: liquid dairy manure digestate (LDMD), liquid dairy manure digestate with ammonia removed (LDMDA-), food waste digestate (FWD), liquid food waste digestate with ammonia removed (LFWDA-), liquid food waste digestate (LFWD), food waste hydrolysate from the Renewable Energy Anaerobic Digester (HREAD), and food waste hydrolysate from the South Area Transfer Station in Sacramento (HSATS). In a red radish (Raphanus sativus) bioassay with H. schachtii, digestates were amended at rates of 0.02, 0.11, 0.57, and 2.86 ml per 100 cm3 of soil. At a rate of 2.86 ml, all amendments except LDMDA- and LFWDA- significantly reduced juvenile root penetration compared with the infested control. In a greenhouse watermelon (Citrullus lanatus) bioassay with Meloidogyne incognita, amendments FWD, LFWD, HREAD, and HSATS as well as LDMD (less effectively) at 2.86 and 5.76 ml per 100 cm3 of soil significantly reduced egg masses per root system compared with the nontreated, nematode-infested control. In a microplot experiment with M. incognita and red radish, in the treatment amended with LFWD at 2.37 ml per 100 cm3 of soil, marketable yields were improved by approximately 50% over the nontreated control and were comparable with those in the treatment with the nematicide Reklemel. In a second microplot experiment with M. incognita and watermelon, treatments that contained LFWD at rates of 3.55 ml per 100 cm3 of soil had transient numerical effects of initial nematode suppression that were not maintained throughout the 3-month growth period. The results of these studies demonstrated that digestates FWD and LFWD consistently expressed some nematode-suppressive capacity.

Dual-directional group delays during optical topological transitions in black phosphorus-based asymmetric hyperbolic metamaterials.

We theoretically study the optical properties of TM waves when their magnetic field direction is perpendicular to the armchair and zigzag optical axes of black phosphorus, respectively. It is found that hyperbolic dispersion and elliptic dispersion coexist in periodically arranged black phosphorus multilayers. Interestingly, by tilting the symmetric multilayers to be asymmetric, the elliptical part of the original two dispersions disappears as the wavelength increases. As such only the hyperbolic dispersion remains, showing an optical topological transition. In the region of the topological transition, a large transmitted group delay (3ps) and a reflected group delay (0.2ps) of the TM waves occurs simultaneously. The corresponding group velocities are slowed down to approximately c/1000 and c/100 (c is the speed of light in a vacuum), respectively. This dual-directional group delays significantly increase the wave-matter interaction so that nonreciprocal perfect absorptions can be realized in the mid-infrared band. Such asymmetrical black phosphorus hyperbolic metamaterials can be applied to the directional, tunable, and nonreciprocal perfect absorbers and also to devices based on strong wave-matter interactions.

Prenatal diagnosis of total anomalous pulmonary venous connection using 2D and HDlive flow combined with spatiotemporal image correlation.

OBJECTIVES: The objective of this study is to examine the application value of two-dimensional (2D) and high-definition live (HDlive) flow combined with spatiotemporal image correlation (STIC) in diagnosing fetal total anomalous pulmonary venous connection (TAPVC). METHODS: Seventeen cases of fetal TAPVC were diagnosed using 2D and HDlive Flow combined with STIC. These cases were then retrospectively analyzed to examine the value of using 2D and HDlive Flow combined with STIC in the diagnosis of TAPVC. RESULTS: 2D and HDlive Flow combined with STIC detected 13 cases of supracardiac TAPVC (two isolated cases, seven cases with right atrial isomerism (RAI), four cases with other complex malformations), one case of isolated intra-cardiac TAPVC, and three cases of cardiac TAPVC (two isolated cases and one case with complex congenital heart anomaly). Small left atrium (LA), the absence of PVs drainage into the LA and the increased retroatrial distance between LA and the descending aorta (DAo) were significant signs that should raise the suspicion of fetal TAPVC. HDlive Flow combined with STIC can dynamically display the TAPVC which may assit the prenatal diagnosis of TAPVC. CONCLUSION: 2D and HDlive Flow combined with STIC can assit the diagnosis of fetal TAPVC abnormalities and has important clinical value.

Process development of sugar beet enzymatic hydrolysis with enzyme recycling for soluble sugar production.

Enzymatic hydrolysis of sugar beets for achieving liquefaction and sugar release is a critical step for beet-ethanol production. An enzyme recycling process was developed in this study to reduce the economic uncertainty raised by the high costs of enzymes by reducing the fresh enzyme usage. A mixture of cellulases and pectinases was used in the beet hydrolysis. The hydrolysate was centrifuged and then processed through a 50 kDa molecular weight cut-off polyethersulfone membrane to recover enzymes from the liquid. Liquid enzyme recycling with 50% fresh enzyme addition achieved a similar liquefaction extent and sugar yield compared to the positive control with 100% fresh enzyme. Solid enzyme recycling showed a lower liquefaction efficiency, requiring at least 75% of fresh enzyme addition for a comparable liquefaction extent. Five sequential batches of hydrolysis with liquid enzyme recycling were successfully conducted to hydrolyze sugar beets with similar liquefaction extents and sugar yields.

Application of biochar and alkalis for recovery of sour anaerobic digesters.

Commercial digesters handling complex waste and organic overloading often encounter unbalanced conditions or failures. With limited studies on the digester recovery from an industry-based waste stream, a complex and high-strength digestate containing up to 79 g COD l-1 from acidified commercial digester was investigated for biochar and alkaline treatments. The addition of biochar and calcium hydroxide successfully decomposed excessive volatile fatty acid up to 18.9 ± 2.5 g l-1 and resumed methane production. The maximum methane yield was obtained from the digester amended with biochar (373.4 ± 6.0 ml g COD-1), followed by calcium hydroxide (350.1 ± 2.5 ml g COD-1). Calcium hydroxide treatment showed a shorter lag phase than the biochar by 44%. Methane production could not be recovered by using sodium hydroxide or untreated digester. This study provides a strategic approach to justify the use of alkalis for restoring sour digesters from industry-based waste streams.

PTPN2 regulates the activation of KRAS and plays a critical role in proliferation and survival of KRAS-driven cancer cells.

RAS genes are the most commonly mutated in human cancers and play critical roles in tumor initiation, progression, and drug resistance. Identification of targets that block RAS signaling is pivotal to develop therapies for RAS-related cancer. As RAS translocation to the plasma membrane (PM) is essential for its effective signal transduction, we devised a high-content screening assay to search for genes regulating KRAS membrane association. We found that the tyrosine phosphatase PTPN2 regulates the plasma membrane localization of KRAS. Knockdown of PTPN2 reduced the proliferation and promoted apoptosis in KRAS-dependent cancer cells, but not in KRAS-independent cells. Mechanistically, PTPN2 negatively regulates tyrosine phosphorylation of KRAS, which, in turn, affects the activation KRAS and its downstream signaling. Consistently, analysis of the TCGA database demonstrates that high expression of PTPN2 is significantly associated with poor prognosis of patients with KRAS-mutant pancreatic adenocarcinoma. These results indicate that PTPN2 is a key regulator of KRAS and may serve as a new target for therapy of KRAS-driven cancer.

Long-term exposure to air pollution and the risk of developing sudden sensorineural hearing loss.

BACKGROUND: The association between exposure to air pollution and sudden sensorineural hearing loss (SSNHL) has not been extensively discussed in the literature. Therefore, we conducted this nationwide study to evaluate the risk of SSNHL in Taiwanese residents with exposure to air pollution. METHODS: We enrolled subjects aged older than 20 years with no history of SSNHL from 1998 to 2010, and followed up until developing SSNHL, withdrawn from the National Health Insurance program, and the end of the database (2011/12/31). The air quality data are managed by Taiwan Environmental Protection Administration. The annual concentrations of PM2.5, SO2, CO, NO, and NO2 from 1998 to 2010 were classified into the three levels according to tertiles. We calculated the annual average of pollutants from baseline until the end of the study, and classified into tertiles. The adjusted hazard ratio (aHR) was estimated by using the multivariate Cox proportional hazard model. RESULTS: When considered continuous air pollutants concentration, subjects who exposed with higher concentration of CO (aHR = 2.16, 95% CI 1.50-3.11), NO (aHR = 1.02, 95% CI 1.01-1.03), and NO2 (aHR = 1.02, 95% CI 1.01-1.04) developing significant higher risk of SSNHL. When classified air pollutants concentration into low, moderate and high level by tertiles, and selected low level as reference, patients exposed with moderate (aHR = 1.56, 95% CI 1.20-2.04) or high level (aHR = 1.33, 95% CI 1.01-1.75) of PM2.5 showed significant higher risk of developing SSNHL. CONCLUSION: This study indicated an increased risk of SSNHL in residents with long-term exposure to air pollution. Nevertheless, further experimental, and clinical studies are needed to validate the study findings.

Chemical and Toxicological Properties of Emissions from a Light-Duty Compressed Natural Gas Vehicle Fueled with Renewable Natural Gas.

Biogas consisting primarily of methane (CH4) and carbon dioxide (CO2) can be upgraded to a transportation fuel referred to as renewable natural gas (RNG) by removing CO2 and other impurities. RNG has energy content comparable to fossil compressed natural gas (CNG) but with lower life-cycle greenhouse gas (GHG) emissions. In this study, a light-duty cargo van was tested with CNG and two RNG blends on a chassis dynamometer in order to compare the toxicity of the resulting exhaust. Tests for reactive oxygen species (ROS), biomarker expressions (CYP1A1, IL8, COX-2), and mutagenicity (Ames) show that RNG exhaust has toxicity that is comparable or lower than CNG exhaust. Statistical analysis reveals associations between toxicity and tailpipe emissions of benzene, dibenzofuran, and dihydroperoxide dimethyl hexane (the last identification is considered tentative/uncertain). Further gas-phase toxicity may be associated with tailpipe emissions of formaldehyde, dimethyl sulfide, propene, and methyl ketene. CNG exhaust contained higher concentrations of these potentially toxic chemical constituents than RNG exhaust in all of the current tests. Photochemical aging of the vehicle exhaust did not alter these trends. These preliminary results suggest that RNG adoption may be a useful strategy to reduce the carbon intensity of transportation fuels without increasing the toxicity of the vehicle exhaust.

A Cascaded Model Based on EfficientDet and YOLACT++ for Instance Segmentation of Cow Collar ID Tag in an Image.

In recent years, many imaging systems have been developed to monitor the physiological and behavioral status of dairy cows. However, most of these systems do not have the ability to identify individual cows because the systems need to cooperate with radio frequency identification (RFID) to collect information about individual animals. The distance at which RFID can identify a target is limited, and matching the identified targets in a scenario of multitarget images is difficult. To solve the above problems, we constructed a cascaded method based on cascaded deep learning models, to detect and segment a cow collar ID tag in an image. First, EfficientDet-D4 was used to detect the ID tag area of the image, and then, YOLACT++ was used to segment the area of the tag to realize the accurate segmentation of the ID tag when the collar area accounts for a small proportion of the image. In total, 938 and 406 images of cows with collar ID tags, which were collected at Coldstream Research Dairy Farm, University of Kentucky, USA, in August 2016, were used to train and test the two models, respectively. The results showed that the average precision of the EfficientDet-D4 model reached 96.5% when the intersection over union (IoU) was set to 0.5, and the average precision of the YOLACT++ model reached 100% when the IoU was set to 0.75. The overall accuracy of the cascaded model was 96.5%, and the processing time of a single frame image was 1.92 s. The performance of the cascaded model proposed in this paper is better than that of the common instance segmentation models, and it is robust to changes in brightness, deformation, and interference around the tag.

Sohlh2 alleviates malignancy of EOC cells under hypoxia via inhibiting the HIF1α/CA9 signaling pathway.

Epithelial ovarian cancer (EOC) is the most common and deadly ovarian cancer. Most of the patients have abdominal/pelvic invasion and metastasis at the time of diagnosis, but the underlying mechanism remains unclear. Insufficiency of blood perfusion and diffusion within most solid tumors can lead to a hypoxic tumor microenvironment and promotes tumor malignancy. In the present study, we detected the role of the spermatogenesis- and oogenesis-specific basic helix-loop-helix (bHLH) transcription factor 2 (sohlh2) on migration, invasion and epithelial-mesenchymal transition (EMT) of EOC cell lines under hypoxia in vitro. We also investigated the possible mechanism underlying it. The results showed that sohlh2 inhibited the migration, invasion and EMT of EOC cells and might function through suppression of the hypoxia-inducible factor 1α (HIF1α)/carbonic anhydrase 9 (CA9) signaling pathway. Our results may open a new avenue for the further development of diagnostic tools and novel therapeutics that will benefit EOC patients.

Visible-Light-Induced Decarboxylative Cyclization of 2-Alkenylarylisocyanides with α-Oxocarboxylic Acids: Access to 2-Acylindoles.

An efficient and practical protocol for visible-light-induced decarboxylative cyclization of 2-alkenylarylisocyanides with α-oxocarboxylic acids has been developed, which afforded a broad range of 2-acylindoles in moderate to good yields. The reaction proceeds through a cascade of acyl radical addition/cyclization reactions under irradiation of an Ir3+ photoredox catalyst without external oxidants and features simple operation, scalability, a broad substrate scope, and good functional group tolerance.

Sohlh2 inhibits breast cancer cell proliferation by suppressing Wnt/ß-catenin signaling pathway.

Sohlh2 belongs to the superfamily of basic helix-loop-helix (bhlh) transcription factors. Aberrant expression of bhlh transcription factors has been shown to be associated with multiple tumorigenesis. We previously identified that sohlh2 functioned as a tumor suppressor in ovarian cancer. Here, we examined the expression levels of sohlh2 in human breast cancer and its potential role in disease pathogenesis. The results of sohlh2 immunohistochemistry (IHC) and Western blot analysis demonstrated the decreased sohlh2 expression in breast cancer specimens as compared to adjacent noncancerous tissues. Through in vitro MTT, BrdU, colony formation and cell cycle assays and in vivo tumor xenograft studies, we showed that forced expression of sohlh2 led to a significant reduction in proliferation due to G1 arrest in vitro and tumorigenesis in nude mice. Conversely, silencing of sohlh2 enhanced breast cancer cell proliferation. Furthermore, we confirmed that sohlh2 inhibited breast cancer cell proliferation by suppressing the Wnt/ß-catenin signaling pathway. APC was the direct target of sohlh2, and mediated the inhibitory activities of sohlh2 on Wnt/ß-catenin signaling pathway. Thus, our data indicate that sohlh2 likely functions as a tumor suppressor in breast cancer that is mediated by repressing Wnt/ß-catenin signaling pathway via upregulation of APC expression.

Cooperation of Dnmt3a R878H with Nras G12D promotes leukemogenesis in knock-in mice: a pilot study.

BACKGROUND: DNMT3A R882H, a frequent mutation in acute myeloid leukemia (AML), plays a critical role in malignant hematopoiesis. Recent findings suggest that DNMT3A mutant acts as a founder mutation and requires additional genetic events to induce full-blown AML. Here, we investigated the cooperation of mutant DNMT3A and NRAS in leukemogenesis by generating a double knock-in (DKI) mouse model harboring both Dnmt3a R878H and Nras G12D mutations. METHODS: DKI mice with both Dnmt3a R878H and Nras G12D mutations were generated by crossing Dnmt3a R878H knock-in (KI) mice and Nras G12D KI mice. Routine blood test, flow cytometry analysis and morphological analysis were performed to determine disease phenotype. RNA-sequencing (RNA-seq), RT-PCR and Western blot were carried out to reveal the molecular mechanism. RESULTS: The DKI mice developed a more aggressive AML with a significantly shortened lifespan and higher percentage of blast cells compared with KI mice expressing Dnmt3a or Nras mutation alone. RNA-seq analysis showed that Dnmt3a and Nras mutations collaboratively caused abnormal expression of a series of genes related to differentiation arrest and growth advantage. Myc transcription factor and its target genes related to proliferation and apoptosis were up-regulated, thus contributing to promote the process of leukemogenesis. CONCLUSION: This study showed that cooperation of DNMT3A mutation and NRAS mutation could promote the onset of AML by synergistically disturbing the transcriptional profiling with Myc pathway involvement in DKI mice.