Proposed risk-scoring model for estimating the prognostic impact of 1q gain in patients with newly diagnosed multiple myeloma.

1q gain (+1q) is the most common high-risk cytogenetic abnormality (HRCA) in patients with multiple myeloma (MM). However, its prognostic value remains unclear in the era of novel agents. Here, we retrospectively analyzed the impact of +1q on the outcomes of 934 patients newly diagnosed with MM. +1q was identified in 53.1% of patients and verified as an independent variate for inferior overall survival (OS) (hazard ratio, 1.400; 95% confidence interval, 1.097-1.787; p = .007). Concurrence of other HRCAs (particularly t(14;16) and del(17p)) further exacerbated the outcomes of patients with +1q, suggesting prognostic heterogeneity. Thus, a risk-scoring algorithm based on four risk variates (t(14;16), hypercalcemia, ISS III, and high LDH) was developed to estimate the outcomes of patients with +1q. Of the patients, 376 evaluable patients with +1q were re-stratified into low (31.6%), intermediate (61.7%), and high risk (6.7%) groups, with significantly different progression-free survival and OS (p < .0001), in association with early relapse of the disease. The prognostic value of this model was validated in the CoMMpass cohort. While attaining undetectable MRD largely circumvented the adverse impact of +1q, it scarcely ameliorated the outcome of the patients with high risk, who likely represent a subset of patients with extremely poor survival. Hence, patients with +1q are a heterogeneous group of high-risk patients, therefore underlining the necessity for their re-stratification. The proposed simple risk-scoring model can estimate the outcomes of patients with +1q, which may help guide risk-adapted treatment for such patients.

The Alpha Subunit of Mitochondrial Processing Peptidase Participated in Fertility Restoration in Honglian-CMS Rice.

The cytoplasmic male sterility (CMS) and nuclear-controlled fertility restoration system is a favorable tool for the utilization of heterosis in plant hybrid breeding. Many restorer-of-fertility (Rf) genes have been characterized in various species over the decades, but more detailed work is needed to investigate the fertility restoration mechanism. Here, we identified an alpha subunit of mitochondrial processing peptidase (MPPA) that is involved in the fertility restoration process in Honglian-CMS rice. MPPA is a mitochondrial localized protein and interacted with the RF6 protein encoded by the Rf6. MPPA indirectly interacted with hexokinase 6, namely another partner of RF6, to form a protein complex with the same molecular weight as the mitochondrial F1F0-ATP synthase in processing the CMS transcript. Loss-of-function of MPPA resulted in a defect in pollen fertility, the mppa+/- heterozygotes showed semi-sterility phenotype and the accumulation of CMS-associated protein ORFH79, showing restrained processing of the CMS-associated atp6-OrfH79 in the mutant plant. Taken together, these results threw new light on the process of fertility restoration by investigating the RF6 fertility restoration complex. They also reveal the connections between signal peptide cleavage and the fertility restoration process in Honglian-CMS rice.

OsGRP3 Enhances Drought Resistance by Altering Phenylpropanoid Biosynthesis Pathway in Rice (Oryza sativa L.).

As a sessile organism, rice often faces various kinds of abiotic stresses, such as drought stress. Drought stress seriously harms plant growth and damages crop yield every year. Therefore, it is urgent to elucidate the mechanisms of drought resistance in rice. In this study, we identified a glycine-rich RNA-binding protein, OsGRP3, in rice. Evolutionary analysis showed that it was closely related to OsGR-RBP4, which was involved in various abiotic stresses. The expression of OsGRP3 was shown to be induced by several abiotic stress treatments and phytohormone treatments. Then, the drought tolerance tests of transgenic plants confirmed that OsGRP3 enhanced drought resistance in rice. Meanwhile, the yeast two-hybrid assay, bimolecular luminescence complementation assay and bimolecular fluorescence complementation assay demonstrated that OsGRP3 bound with itself may affect the RNA chaperone function. Subsequently, the RNA-seq analysis, physiological experiments and histochemical staining showed that OsGRP3 influenced the phenylpropanoid biosynthesis pathway and further modulated lignin accumulation. Herein, our findings suggested that OsGRP3 enhanced drought resistance in rice by altering the phenylpropanoid biosynthesis pathway and further increasing lignin accumulation.

Identification and Allele Combination Analysis of Rice Grain Shape-Related Genes by Genome-Wide Association Study.

Grain shape is an important agronomic character of rice, which affects the appearance, processing, and the edible quality. Screening and identifying more new genes associated with grain shape is beneficial to further understanding the genetic basis of grain shape and provides more gene resources for genetic breeding. This study has a natural population containing 623 indica rice cultivars. Genome-wide association studies/GWAS of several traits related to grain shape (grain length/GL, grain width/GW, grain length to width ratio/GLWR, grain circumferences/GC, and grain size/grain area/GS) were conducted by combining phenotypic data from four environments and the second-generation resequencing data, which have identified 39 important Quantitative trait locus/QTLs. We analyzed the 39 QTLs using three methods: gene-based association analysis, haplotype analysis, and functional annotation and identified three cloned genes (GS3, GW5, OsDER1) and seven new candidate genes in the candidate interval. At the same time, to effectively utilize the genes in the grain shape-related gene bank, we have also analyzed the allelic combinations of the three cloned genes. Finally, the extreme allele combination corresponding to each trait was found through statistical analysis. This study's novel candidate genes and allele combinations will provide a valuable reference for future breeding work.

UBM-guided scleral buckling for Schwartz-Matsuo syndrome with tear of nonpigmented epithelium of the ciliary body: a case report.

BACKGROUND: Tears in Schwartz-Matsuo syndrome are generally confirmed by preoperative ophthalmoscopic examination. A case of Schwartz-Matsuo syndrome with a tear detected by ultrasound biomicroscopy (UBM) and treated by UBM-guided scleral buckling was reported, and its mechanism was analysed. CASE PRESENTATION: A 40-year-old Chinese man presented with blurry vision and intermittent eye pain in his left eye for three days. The visual acuity of the left eye decreased from 20/20 to 20/40, and the intraocular pressure (IOP) fluctuated dramatically from 24.0 mmHg to 56.7 mmHg at the first visit. Gonioscopy revealed that the chamber angle remained open. A macula-involving inferior retinal detachment extending from 4:30 to 9:30 with no obvious causative break was observed through ophthalmoscopic examination. However, a single small tear was detected at the nonpigmented epithelium of pars plana of the ciliary body at approximately 7-8 o'clock by UBM. The loss of photoreceptor outer segments and ellipsoid zone and the existence of macular microcysts in the inner and outer nuclear layers were observed in the detached macula by optical coherence tomography. Then, he underwent successful scleral buckling guided by UBM. Three months later, the retina was flat with normal IOP, and the best corrected visual acuity of his left eye gradually improved to 20/25. UBM confirmed the closure of the tear. CONCLUSIONS: Tear of the nonpigmented epithelium of the ciliary body is a rare condition associated with Schwartz-Matsuo syndrome. UBM plays a key role in detecting occult tears of the nonpigmented epithelium of the ciliary body, guiding scleral buckling surgery, and observing the closure of the tear postoperatively.

Integrative Transcriptomic and Proteomic Analysis Reveals an Alternative Molecular Network of Glutamine Synthetase 2 Corresponding to Nitrogen Deficiency in Rice (Oryza sativa L.).

Nitrogen (N) is an essential nutrient for plant growth and development. The root system architecture is a highly regulated morphological system, which is sensitive to the availability of nutrients, such as N. Phenotypic characterization of roots from LY9348 (a rice variety with high nitrogen use efficiency (NUE)) treated with 0.725 mM NH4NO3 (1/4N) was remarkable, especially primary root (PR) elongation, which was the highest. A comprehensive analysis was performed for transcriptome and proteome profiling of LY9348 roots between 1/4N and 2.9 mM NH4NO3 (1N) treatments. The results indicated 3908 differential expression genes (DEGs; 2569 upregulated and 1339 downregulated) and 411 differential abundance proteins (DAPs; 192 upregulated and 219 downregulated). Among all DAPs in the proteome, glutamine synthetase (GS2), a chloroplastic ammonium assimilation protein, was the most upregulated protein identified. The unexpected concentration of GS2 from the shoot to the root in the 1/4N treatment indicated that the presence of an alternative pathway of N assimilation regulated by GS2 in LY9348 corresponded to the low N signal, which was supported by GS enzyme activity and glutamine/glutamate (Gln/Glu) contents analysis. In addition, N transporters (NRT2.1, NRT2.2, NRT2.3, NRT2.4, NAR2.1, AMT1.3, AMT1.2, and putative AMT3.3) and N assimilators (NR2, GS1;1, GS1;2, GS1;3, NADH-GOGAT2, and AS2) were significantly induced during the long-term N-deficiency response at the transcription level (14 days). Moreover, the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis demonstrated that phenylpropanoid biosynthesis and glutathione metabolism were significantly modulated by N deficiency. Notably, many transcription factors and plant hormones were found to participate in root morphological adaptation. In conclusion, our study provides valuable information to further understand the response of rice roots to N-deficiency stress.

IPA1 Negatively Regulates Early Rice Seedling Development by Interfering with Starch Metabolism via the GA and WRKY Pathways.

Ideal Plant Architecture 1 (IPA1) encodes SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 14 (SPL14) with a pleiotropic effect on regulating rice development and biotic stress responses. To investigate the role of IPA1 in early seedling development, we developed a pair of IPA1/ipal-NILs and found that seed germination and early seedling growth were retarded in the ipa1-NIL. Analysis of the soluble sugar content, activity of amylase, and expression of the α-amylase genes revealed that the starch metabolism was weakened in the ipa1-NIL germinating seeds. Additionally, the content of bioactive gibberellin (GA) was significantly lower than that in the IPA1-NIL seeds at 48 h of imbibition. Meanwhile, the expression of GA synthesis-related gene OsGA20ox1 was downregulated, whereas the expression of GA inactivation-related genes was upregulated in ipa1-NIL seeds. In addition, the expression of OsWRKY51 and OsWRKY71 was significantly upregulated in ipa1-NIL seeds. Using transient dual-luciferase and yeast one-hybrid assays, IPA1 was found to directly activate the expression of OsWRKY51 and OsWRKY71, which would interfere with the binding affinity of GA-induced transcription factor OsGAMYB to inhibit the expression of α-amylase genes. In summary, our results suggest that IPA1 negatively regulates seed germination and early seedling growth by interfering with starch metabolism via the GA and WRKY pathways.

Comparative transcriptome analysis and identification of candidate adaptive evolution genes of Miscanthus lutarioriparius and Miscanthus sacchariflorus.

Miscanthus species are perennial C4 grasses that are considered promising energy crops because of their high biomass yields, excellent adaptability and low management costs. Miscanthus lutarioriparius and Miscanthus sacchariflorus are closely related subspecies that are distributed in different habitats. However, there are only a few reports on the mechanisms by which Miscanthus adapts to different environments. Here, comparative transcriptomic and morphological analyses were used to study the evolutionary adaptation of M. lutarioriparius and M. sacchariflorus to different habitats. In total, among 7586 identified orthologs, 2060 orthologs involved in phenylpropanoid biosynthesis and plant hormones were differentially expressed between the two species. Through an analysis of the Ka/Ks ratios of the orthologs, we estimated that the divergence time between the two species was approximately 4.37 Mya. In addition, 37 candidate positively selected orthologs (PSGs) that played important roles in the adaptation of these species to different habitats were identified. Then, the expression levels of 20 PSGs in response to flooding and drought stress were analyzed, and the analysis revealed significant changes in their expression levels. These results facilitate our understanding of the evolutionary adaptation to habitats and the speciation of M. lutarioriparius and M. sacchariflorus. We hypothesise that lignin synthesis genes are the main cause of the morphological differences between the two species. In summary, the plant nonspecific phospholipase C gene family and the receptor-like protein kinase gene family played important roles in the evolution of these two species. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01030-1.

Comparative population genomics reveals genetic divergence and selection in lotus, Nelumbo nucifera.

BACKGROUND: Lotus (Nelumbo nucifera) is an aquatic plant with important agronomic, horticulture, art and religion values. It was the basal eudicot species occupying a critical phylogenetic position in flowering plants. After the domestication for thousands of years, lotus has differentiated into three cultivated types -flower lotus, seed lotus and rhizome lotus. Although the phenotypic and genetic differentiations based on molecular markers have been reported, the variation on whole-genome level among the different lotus types is still ambiguous. RESULTS: In order to reveal the evolution and domestication characteristics of lotus, a total of 69 lotus accessions were selected, including 45 cultivated accessions, 22 wild sacred lotus accessions, and 2 wild American lotus accessions. With Illumina technology, the genomes of these lotus accessions were resequenced to > 13× raw data coverage. On the basis of these genomic data, 25 million single-nucleotide polymorphisms (SNPs) were identified in lotus. Population analysis showed that the rhizome and seed lotus were monophyletic and genetically homogeneous, whereas the flower lotus was biphyletic and genetically heterogeneous. Using population SNP data, we identified 1214 selected regions in seed lotus, 95 in rhizome lotus, and 37 in flower lotus. Some of the genes in these regions contributed to the essential domestication traits of lotus. The selected genes of seed lotus mainly affected lotus seed weight, size and nutritional quality. While the selected genes were responsible for insect resistance, antibacterial immunity and freezing and heat stress resistance in flower lotus, and improved the size of rhizome in rhizome lotus, respectively. CONCLUSIONS: The genome differentiation and a set of domestication genes were identified from three types of cultivated lotus- flower lotus, seed lotus and rhizome lotus, respectively. Among cultivated lotus, flower lotus showed the greatest variation. The domestication genes may show agronomic importance via enhancing insect resistance, improving seed weight and size, or regulating lotus rhizome size. The domestication history of lotus enhances our knowledge of perennial aquatic crop evolution, and the obtained dataset provides a basis for future genomics-enabled breeding.

IRF2-INPP4B axis inhibits apoptosis of acute myeloid leukaemia cells via regulating T helper 1/2 cell differentiation.

The interferon-regulatory factor 2 (IRF2)-inositol polyphosphate-4-phosphatase type-II (INPP4B) axis has been shown to suppress cell apoptosis by inducing autophagy in acute myeloid leukaemia (AML) cells. T helper type 1 cell (Th1)/Th2 imbalance is involved in development of autophagy and AML. The aim of this study was to investigate whether IRF2-INPP4B axis regulates autophagy and apoptosis of AML cells via regulating Th1/Th2 cell differentiation. The frequencies of Th2 cells and Th1 cells in transfected CD4+ T cells were determined by flow cytometry. The levels of TNF-α, IFN-γ, IL-4, and IL-13 in peripheral blood and transfected CD4+ T cells from AML patients and healthy donors were examined by ELISA assay. The mRNA levels of IRF2 and INPP4B in peripheral blood mononuclear cells (PBMCs) from AML patients and healthy donors were detected by qRT-PCR. Autophagy was evaluated by green fluorescent protein (GFP)-LC3 immunofluorescence and western blot analysis of autophagy-associated proteins. AML cell apoptosis was detected by flow cytometry. In this study, elevated frequencies of Th2 cells and reduced frequencies of Th1 cells, as well as higher expression of IRF2 and INPP4B, were observed in PBMCs from AML patients relative to healthy donors. Furthermore, IRF2 inhibited Th1 cell differentiation and promoted Th2 cell differentiation through INPP4B. Moreover, we confirmed that IRF2-INPP4B-mediated regulation of Th1/Th2 differentiation promoted autophagy and inhibited apoptosis of AML cells. Collectively, IRF2-INPP4B axis regulates autophagy and apoptosis of AML cells via regulating Th1/Th2 cell differentiation. SIGNIFICANCE OF THE STUDY: In the present study, we confirmed that IRF2-INPP4B-mediated regulation of Th1/Th2 balance promoted autophagy and inhibited apoptosis of AML cells. These findings might provide clues in better understanding of the mechanisms of AML.

In situ optical spectroscopy characterization for optimal design of lithium-sulfur batteries.

The lithium-sulfur (Li-S) battery is one of the most promising high-energy-density secondary battery systems. However, it suffers from issues arising from its extremely complicated "solid-liquid-solid" reaction routes. In recent years, enormous advances have been made in optimizing Li-S batteries via the rational design of compositions and architectures. Nevertheless, a comprehensive and in-depth understanding of the practical reaction mechanisms of Li-S systems and their effect on the electrochemical performance is still lacking. Very recently, several important in situ optical spectroscopic techniques, including Raman, infrared and ultraviolet-visible spectroscopies, have been developed to monitor the real-time variations of the battery states, and a bridge linking the macroscopic electrochemical performance and microscopic architectures of the components has been set up, thus playing a critical role in scientifically guiding further optimal design of Li-S batteries. In this tutorial review, we provide a systematic summary of the state-of-the-art innovations in the characterization and optimal design of Li-S batteries with the aid of these in situ optical spectroscopic techniques, to guide a beginner to construct in situ optical spectroscopy electrochemical cells, and develop strategies for preventing long-chain polysulfide formation, dissolution and migration, thus alleviating the shuttle effect in Li-S batteries and improving the cell performances significantly.

Transcriptome, physiological and biochemical analysis of Triarrhena sacchariflora in response to flooding stress.

BACKGROUND: In recent decades, the frequency of flooding is increasing with the change of global climate. Flooding has become one of the major abiotic stresses that seriously affect growth and development of plants. Triarrhena sacchariflora Nakai has been considered a promising energy crop for utilization in ethanol production. Flooding stress is among the most severe abiotic stressors in the production of Nakai. However, the physiological and molecular biological mechanisms of Nakai response to flooding is still unclear. In the present study, in order to understand the molecular mechanisms of Nakai in response to flooding stress, the transcriptome, physiological and biochemical were investigated. RESULTS: The results demonstrated that significant physiological changes were observed in photosynthetic system, antioxidative enzyme activity, chlorophyll, carotenoid, proline, lipid peroxidation and soluble sugar content under normal and flooding treatments. Such as, the chlorophyll, carotenoid contents and photosynthetic system were significantly decreased. Whereas, the antioxidative enzyme activity, proline, lipid peroxidation and soluble sugar has increased first and then decreased under treatments compared with the normal plants. Additionally, a total of 8832, 6608 and 3649 unigenes were validated to be differentially expressed under different treatments, respectively. Besides, gene ontology enrichment and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis of the different expression levels of genes also presented processes, which involved in photosynthesis, sucrose catabolism, glycolysis, stress response and defense, phytohormone biosynthesis and signal transduction. CONCLUSIONS: The results provide a comprehensive view of the complex molecular events involved in the response to flooding stress of Nakai leaves, which also will promote the research in the development of flood-resistant crops and provide new tools for Nakai breeders.

Selection of suitable reference genes for quantitive real-time PCR normalization in Miscanthus lutarioriparia.

Miscanthus lutarioriparia, which is found widespread in China, has attracted great attention as a most potential bioenergy plant for years. The quantitative real time PCR (RT-qPCR) has appeared as a sensitive and powerful technique to measure gene expression in living organisms during different development stages. In this study, we evaluated ten candidate genes, including 25S ribosomal RNA gene (25S rRNA), actin1 gene (ACT1), carotenoid-binding protein 20 gene (CBP20), glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH), Ubiquitin gene (UBQ), eukaryotic elongation factor 1-αgene (eEF-1α), α-tubulin gene (α-TUB), ß-tubulin gene (ß-TUB), eukaryotic translation initiation factor 4α-1 gene (eIF-4α) and NAC domain protein gene(NAC) in a series of 30 M. lutarioriparia samples followed by statistical algorithms geNorm and Normfinder to analyze the gene expression stability. The results indicated that eIF-4αand UBQ were the most stable expressed genes while CBP20 showed as the least stable among all the samples. Based on above research, we recommend that at least two top-ranked reference genes should be employed for expression data normalization. The best genes selected in this study will provide a starting point to select reference genes in the future in other tissues and under other experimental conditions in this energy crop candidate.

OsPRR37 confers an expanded regulation of the diurnal rhythms of the transcriptome and photoperiodic flowering pathways in rice.

The circadian clock enables organisms to rapidly adapt to the ever-changing environmental conditions that are caused by daily light/dark cycles. Circadian clock genes universally affect key agricultural traits, particularly flowering time. Here, we show that OsPRR37, a circadian clock gene, delays rice flowering time in an expression level-dependent manner. Using high-throughput mRNA sequencing on an OsPRR37 overexpressing transgenic line (OsPRR37-OE5) and the recipient parent Guangluai4 that contains the loss-of-function Osprr37, we identify 14,992 genes that display diurnal rhythms, which account for 52.9% of the transcriptome. Overexpressing OsPRR37 weakens the transcriptomic rhythms and alters the phases of rhythmic genes. In total, 3,210 differentially expressed genes (DEGs) are identified, among which 1,863 rhythmic DEGs show a correlation between the change of absolute amplitudes and the mean expression levels. We further reveal that OsPRR37 functions as a transcriptional repressor to repress the expression levels and amplitudes of day-phased clock genes. More importantly, OsPRR37 confers expanded regulation on the evening-phased rhythmic DEGs by repressing the morning-phased rhythmic DEGs. Further study shows that OsPRR37 expands its regulation on flowering pathways by repressing Ehd1. Thus, our results demonstrate an expanded regulation mechanism of the circadian clock on the diurnal rhythms of the transcriptome.

Molecular cloning and characterization of two manganese superoxide dismutases from Miscanthus × giganteus.

KEY MESSAGE: Six MnSOD genes were isolated from five Miscanthus species. MgMnSOD1 functions in mitochondria and MgMnSOD1 seems to be the main MnSOD gene involved in stress response of M. × giganteus. Miscanthus × giganteus is a promising biomass energy crop with advantages of vigorous growth, high yield, low fertilizer and pesticide inputs. However, poor overwinter ability limits its widespread cultivation. Moreover, narrow genetic base may increase the risk of susceptibility to diseases and pests. Manganese superoxide dismutase (MnSOD), an important antioxidant enzyme involved in stress tolerance is able to protect plant cells from accumulated reactive oxygen species by converting superoxide to peroxide and oxygen. In many plants, overexpression of MnSOD has shown the ability to enhance the resistance to various stresses. This article describes the studies performed in an attempt to elucidate the molecular and enzymatic properties of MnSODs in M. × giganteus. MnSOD genes from M. × giganteus (MgMnSOD1, MgMnSOD2), M. lutarioriparia (MlMnSOD), M. sacchariflora (MsaMnSOD), M. sinensis (MsiMnSOD), and M. floridulus (MfMnSOD) were cloned and sequenced. The sequence analysis and expression patterns of MgMnSOD1 and MgMnSOD2 suggest that they were orthologous genes which were inherited from the two parents, M. sacchariflora and M. sinensis, respectively. In addition, MgMnSOD1 is predicted to be the main MnSOD gene involved in stress response of M. × giganteus. The activity of purified recombinant MgMnSOD1 was 1854.79 ± 39.98 U mg(-1) (mean ± SD). Further enzymatic assays revealed that the protein exhibited an outstanding thermal stability. MgMnSOD1 is predicted to be targeted to mitochondria and involved in removing the superoxide radical generated by respiration. The presence and sequences of other SOD isozymes transcripts were also investigated in this study.

Cloning a glutathione peroxidase gene from Nelumbo nucifera and enhanced salt tolerance by overexpressing in rice.

A full-length cDNA clone encoding an 866 bp-length glutathione peroxidase protein (NnGPX) was isolated from lotus (Nelumbo nucifera L.). The deduced amino acid sequence of the NnGPX gene had significant homology with ATGPX6. A 3D structural model of the NnGPX was constructed by homology modeling. The cloned NnGPX gene was expressed in Escherichia coli, and a fusion protein of about 40 kDa was detected after isopropyl thiogalactoside induction. Under different concentrations of Na2SeO3 treatments, NnGPX was found to be an enzyme that does not contain selenium. Real-time PCR analysis showed that the NnGPX gene was expressed in all organs of lotus, and its high expression mainly occurred in organs with active metabolisms. NnGPX transcript increased remarkably in response to cold, heat, mechanical damage, and salt treatment. Subsequently, the NnGPX gene was introduced in Oryza sativa cv. Yuetai B. PCR results verified the integration of this gene into the genome of rice and reverse transcription-PCR verified that this gene had been expressed in transgenic rice. The transgenic plants were significantly more tolerant to salt stress compared with the wild-type.

Genetic mutations and immune microenvironment: unveiling the connection to AML prognosis.

BACKGROUND: This study aims to investigate the correlation between NK and NKT cell proportion disparities and prognosis in patients with acute myeloid leukemia (AML). METHODS: Forty-four cases of acute myeloid leukemia patients were selected, and flow cytometry was utilized to evaluate the expression of bone marrow NK and NKT cells. Next-generation sequencing technology was employed to detect genetic mutations in these 44 AML patients, and the rates of first induction remission and overall survival were recorded. Comparisons were made to analyze the respective differences in NK and NKT cell proportions among AML patients with various genetic mutations and risk stratifications. RESULTS: The FLT-3-ITD+ group exhibited a significant increase in the proportion of NK cells compared to the normal control group and FLT3-ITD+/NPM1+ group, whereas the proportion of NKT cells was significantly decreased. Additionally, the CEBPA+ group showed an increased proportion of NKT cells compared to the TP53+ group and ASXL1+ group. The high-risk group had a higher proportion of NK cells than the intermediate-risk group, while the proportion of NKT cells was lower in the high-risk group compared to the intermediate-risk group.Patients achieving first induction remission displayed a higher proportion of NKT cells at initial diagnosis compared to those who did not achieve remission. The distribution of NK cells show significant differences among AML patients in different survival periods. CONCLUSION: This results implies that distinct genetic mutations may play a role not only in tumor initiation but also in shaping the tumor microenvironment, consequently impacting prognosis.

Evaluation and validation of suitable reference genes for quantitative real-time PCR analysis in lotus (Nelumbo nucifera Gaertn.).

The qRT-PCR technique has been regarded as an important tool for assessing gene expression diversity. Selection of appropriate reference genes is essential for validating deviation and obtaining reliable and accurate results. Lotus (Nelumbo nucifera Gaertn) is a common aquatic plant with important aesthetic, commercial, and cultural values. Twelve candidate genes, which are typically used as reference genes for qRT-PCR in other plants, were selected for this study. These candidate reference genes were cloned with, specific primers designed based on published sequences. In particular, the expression level of each gene was examined in different tissues and growth stages of Lotus. Notably, the expression stability of these candidate genes was assessed using the software programs geNorm and NormFinder. As a result, the most efficient reference genes for rootstock expansion were TBP and UBQ. In addition, TBP and EF-1α were the most efficient reference genes in various floral tissues, while ACT and GAPDH were the most stable genes at all developmental stages of the seed. CYP and GAPDH were the best reference genes at different stages of leaf development, but TUA was the least stable. Meanwhile, the gene expression profile of NnEXPA was analyzed to confirm the validity of the findings. It was concluded that, TBP and GAPDH were identified as the best reference genes. The results of this study may help researchers to select appropriate reference genes and thus obtain credible results for further quantitative RT-qPCR gene expression analyses in Lotus.