Targeted gene sequencing and transcriptome sequencing reveal characteristics of NUP98 rearrangement in pediatric acute myeloid leukemia.

BACKGROUND: NUP98 rearrangements (NUP98-r) are rare but overrepresented mutations in pediatric acute myeloid leukemia (AML) patients. NUP98-r is often associated with chemotherapy resistance and a particularly poor prognosis. Therefore, characterizing pediatric AML with NUP98-r to identify aberrations is critically important. METHODS: Here, we retrospectively analyzed the clinicopathological features, genomic and transcriptomic landscapes, treatments, and outcomes of pediatric patients with AML. RESULTS: Nine patients with NUP98-r mutations were identified in our cohort of 142 patients. Ten mutated genes were detected in patients with NUP98-r. The frequency of FLT3-ITD mutations differed significantly between the groups harboring NUP98-r and those without NUP98-r (P = 0.035). Unsupervised hierarchical clustering via RNA sequencing data from 21 AML patients revealed that NUP98-r samples clustered together, strongly suggesting a distinct subtype. Compared with that in the non-NUP98-r fusion and no fusion groups, CMAHP expression was significantly upregulated in the NUP98-r samples (P < 0.001 and P = 0.001, respectively). Multivariate Cox regression analyses demonstrated that patients harboring NUP98-r (P < 0.001) and WT1 mutations (P = 0.030) had worse relapse-free survival, and patients harboring NUP98-r (P < 0.008) presented lower overall survival. CONCLUSIONS: These investigations contribute to the understanding of the molecular characteristics, risk stratification, and prognostic evaluation of pediatric AML patients.

Trivalent nanobody-based ligands mediate powerful activation of GPVI, CLEC-2, and PEAR1 in human platelets whereas FcγRIIA requires a tetravalent ligand.

BACKGROUND: Clustering of the receptors glycoprotein receptor VI (GPVI), C-type lectin-like receptor 2 (CLEC-2), low-affinity immunoglobulin γ Fc region receptor II-a (FcγRIIA), and platelet endothelial aggregation receptor 1 (PEAR1) leads to powerful activation of platelets through phosphorylation of tyrosine in their cytosolic tails and initiation of downstream signaling cascades. GPVI, CLEC-2, and FcγRIIA signal through YxxL motifs that activate Syk. PEAR1 signals through a YxxM motif that activates phosphoinositide 3-kinase. Current ligands for these receptors have an undefined valency and show significant batch variation and, for some, uncertain specificity. OBJECTIVES: We have raised nanobodies against each of these receptors and multimerized them to identify the minimum number of epitopes to achieve robust activation of human platelets. METHODS: Divalent and trivalent nanobodies were generated using a flexible glycine-serine linker. Tetravalent nanobodies utilize a mouse Fc domain (IgG2a, which does not bind to FcγRIIA) to dimerize the divalent nanobody. Ligand affinity measurements were determined by surface plasmon resonance. Platelet aggregation, adenosine triphosphate secretion, and protein phosphorylation were analyzed using standardized methods. RESULTS: Multimerization of the nanobodies led to a stepwise increase in affinity with divalent and higher-order nanobody oligomers having sub-nanomolar affinity. The trivalent nanobodies to GPVI, CLEC-2, and PEAR1 stimulated powerful and robust platelet aggregation, secretion, and protein phosphorylation at low nanomolar concentrations. A tetravalent nanobody was required to activate FcγRIIA with the concentration-response relationship showing a greater variability and reduced sensitivity compared with the other nanobody-based ligands, despite a sub-nanomolar binding affinity. CONCLUSION: The multivalent nanobodies represent a series of standardized, potent agonists for platelet glycoprotein receptors. They have applications as research tools and in clinical assays.

The effect of the plasma methotrexate concentration during high-dose methotrexate therapy in childhood acute lymphoblastic leukemia.

Two hundred and thirty-one acute lymphoblastic leukemia (ALL) children with 1376 high-dose methotrexate (HD-MTX) courses (3-5 g/m2) were enrolled to analyze the influence of the plasma MTX concentration (CMTX) in ALL. The 24-h target peak CMTX (C24h) was set at 33 µmol/l for low-risk (LR) and 65 µmol/l for intermediate/high-risk (IR/HR) groups. The median C24h was 42.0 µmol/l and 69.7 µmol/l for LR and IR/HR groups, respectively. MTX excretion delay was observed in 14.6% of courses, which was more frequent in IR/HR groups (56.9% vs. LR group 40.2%, p = .014) and T-ALL patients (82.6% vs. B-ALL 47.1%, p = .001). MTX-related toxicities were more common in courses with MTX excretion delay. However, survival between the patients who failed to reach the target C24h or not, with or without MTX excretion delay, was comparable. These findings suggest that, owing to the effectiveness of risk stratification chemotherapy, CMTX does not exert an independent influence on the prognosis of childhood ALL.

The predictive utility of cytokines, procalcitonin and C-reactive protein among febrile pediatric hematology and oncology patients with severe sepsis or septic shock.

Severe sepsis and septic shock are life-threatening for pediatric hematology and oncology patient receiving chemotherapy. Th1/Th2 cytokines, C-reactive protein (CRP), and procalcitonin (PCT) are all thought to be associated with disease severity. The aim of this study was to prospectively verify the utility of Th1/Th2 cytokines and compare them with PCT and CRP in the prediction of adverse outcomes. Data on patients were collected from January 1, 2011, to December 31, 2020. Blood samples were taken for Th1/Th2 cytokine, CRP, and PCT measurements at the initial onset of infection. Severe infection (SI) was defined as severe sepsis or septic shock. Th1/Th2 cytokine levels were determined by using flow cytometric bead array technology. In total, 7,735 febrile episodes were included in this study. For SI prediction, the AUCs of IL-6, IL-10 and TNF-α were 0.814, 0.805 and 0.624, respectively, while IL-6 and IL-10 had high sensitivity and specificity. IL-6 > 220.85 pg/ml and IL-10 > 29.95 pg/ml had high odds ratio (OR) values of approximately 3.5 in the logistic regression. Within the subgroup analysis, for bloodstream infection (BSI) prediction, the AUCs of IL-10 and TNF-α were 0.757 and 0.694, respectively. For multiorgan dysfunction syndrome (MODS) prediction, the AUC of CRP was 0.606. The AUC of PCT for mortality prediction was 0.620. In conclusion, IL-6 and IL-10 provide good predictive value for the diagnosis of SI. For children with SI, IL-10 and TNF-α are associated with BSI, while CRP and PCT are associated with MODS and death, respectively.

Terpenoids from Euphorbia helioscopia and Their Cytotoxic Activities against H1975 Cells.

Three previously undescribed diterpenoids, helioscopnoids A-C, and eight known compounds were isolated from the whole plants of Euphorbia helioscopia. Their structures were established by extensive analysis of spectra and data comparison with previous literatures. Among them, compound 4 was identified as 24,24-dimethoxy-25,26,27-trinoreuphan-3ß-ol with revised configurations of C-13, C-14, and C-17 (13R*, 14R*, 17R*). Cytotoxicity assays revealed that all compounds exhibited varying levels of cytotoxicity against H1975 cells, with compound 9 displaying the most potent activity, as indicated by cell viability rates of 18.13 % and 20.76 % at concentrations of 20 µM and 5 µM, respectively. This study expands the understanding of E. helioscopia terpenoids' structural diversity and biological activities, contributing to the exploration of potential therapeutic applications.

Thromboelastography parameters in chronic viral liver disease and liver resection: A retrospective study.

Introduction: Thromboelastography (TEG) provides a global assessment of haemostasis and is potentially applicable to liver disease. The present study aimed to explore the utility of TEG for the evaluation of patients with chronic viral liver disease, which has previously not been investigated. Methods: Demographic characteristics and TEG parameters were collected before surgery. Child-Turcotte-Pugh (CTP) and model for end-stage liver disease (MELD) scores were used to categorise stages of liver cirrhosis. Liver resections were classified as low, medium and high complexity. Results: A total of 344 patients were included. Results showed significantly longer K-time, smaller α-angle and lower maximum amplitude (MA) with increasing liver disease severity as measured by the CTP and MELD scores (P < 0.05 for all). After multivariable adjustment (including age, sex, liver disease aetiology, alanine aminotransferase [ALT], aspartate aminotransferase [AST], albumin, total bilirubin, haemoglobin and platelet count), TEG parameters (except R-times) were either weakly or inversely related to the severity of liver disease as defined by the MELD score (absolute r < 0.2 and P < 0.05 for all except R-times). R-times obtained before surgery were weakly correlated with perioperative blood loss (r < 0.2 and P < 0.05 for all). Conclusions: The correlation between TEG parameters and severity of liver disease was weak. In addition, R-times obtained before liver resection were weakly associated with perioperative blood loss after multivariable adjustments. TEG utility for haemostasis assessment and prediction of blood loss during liver resection should be further explored in high-quality studies.

Atypical 'long-tailed' cockroaches arose during Cretaceous in response to angiosperm terrestrial revolution.

Typical cockroaches are flat, broad, with large pronotum and wings covering the body. This conserved morphotype dates back to the Carboniferous, during which the ancestral cockroaches, or roachoids, originated. On the other hand, the ovipositor of cockroaches gradually reduced during the Mesozoic, coupled with a major shift of reproductive strategy. By the Cretaceous, long external ovipositors became rare, most cockroaches used very short or even hidden internal ovipositors to fabricate egg cases (oothecae), which is an innovation for egg protection. Here, we describe two cockroaches from mid-Cretaceous Myanmar amber: Ensiferoblatta oecanthoides gen. et sp. nov. (Ensiferoblattidae fam. nov.) and Proceroblatta colossea gen. et sp. nov. They are slim, elongate, fusiform, with longitudinal pronotum, and have long external ovipositors. The combination of these traits represents a unique morphotype, which resembles crickets and katydids (Ensifera) more than general cockroaches. Ensiferoblatta and Proceroblatta may be arboreal, feeding on and/or laying eggs into certain angiosperms that newly emerged. Their open habit causes latent impairment to viability, and may contribute to their extinction. These new taxa are the youngest members of the ancient, extinct group of cockroaches, namely Eoblattodea, which are characterized by long ovipositors. We speculate that the extinction of certain gymnosperm hosts almost ended the 200-My triumph of Eoblattodea. Despite an attempt to adapt to angiosperm hosts, Ensiferoblatta, Proceroblatta and suchlike cockroaches as an evolutionary dead end failed to save Eoblattodea from extinction. The lack of protection for eggs (maternal care in particular) might accelerate the extinction of Eoblattodea as a whole.

A new genus of japygids (Diplura: Japygidae) in mid-Cretaceous amber from northern Myanmar.

Diplura is a cryptic and edaphic group of hexapod animals characterized by the presence of filiform or pincer-like cerci located at the end of the abdomen. As one of the early diverging lineages of Hexapoda, diplurans play a crucial role in unraveling the origin of the hyperdiverse insects. The fossil record of diplurans, however, is exceptionally limited due to their delicate bodies and habitat requirements. Here we report the discovery of a new japygid, Cretojapyx huangi gen. et sp. nov., entombed in mid-Cretaceous Kachin amber (northern Myanmar). This specimen exhibits distinctive features, including the well-developed carinae on abdominal segment X and cerci, left cercus lacking a prominent tooth and denticles, and right cercus with a postmedian tooth followed by uniseriate round denticles. Our finding provides a unique opportunity to gain valuable insights into the evolutionary history of this ancient lineage and deepen our understanding of terrestrial ecosystems during the Mesozoic.

A new marsh beetle from mid-Cretaceous amber of northern Myanmar (Coleoptera: Scirtidae).

As one of the earliest-diverging lineage of the megadiverse beetle suborder Polyphaga, marsh beetles (Scirtidae) are crucial for reconstructing the ancestor of all polyphagan beetles and the ecomorphological underpinnings of their remarkable evolutionary success. The phylogeny of marsh beetles has nonetheless remained challenging to infer, not least because of their fragmentary Mesozoic fossil record. Here we describe a new scirtid beetle genus and species, Varcalium lawrencei gen. et sp. nov., preserving internal tissue, from Albian-Cenomanian Kachin amber (ca 99 Ma), representing the second member of this family known from the deposit. Based on a formal morphological phylogenetic analysis, Varcalium is recovered within the crown-group of Scirtinae, forming a clade with other genera that possess subocular carinae. The finding suggests that the crown-group of Scirtinae has already diversified by the mid-Cretaceous.

[Value of red blood cell distribution width in evaluating the severity of illness of novel coronavirus Delta variant].

OBJECTIVE: To explore the value of red blood cell distribution width (RDW) in evaluating the severity of patients infected with novel coronavirus Delta variant. METHODS: A total of 28 patients infected with novel coronavirus Delta variant in designated hospital treated by the First Affiliated Hospital of Xi'an Jiaotong University medical team from December 2021 to January 2022 were enrolled (23 cases of common type, 4 severe and 1 critical cases). The detailed clinical data of patients was collected. Then, Pearson's correlation analysis was used to identify the blood examination indexes which affected the arterial partial pressure of oxygen (PaO2) and arterial partial pressure of carbon dioxide (PaCO2). According to the median standard deviation of red blood cell distribution width (RDW-SD, 42.5 fL), 28 patients were divided into low RDW-SD group (≤ 42.5 fL, 16 cases) and high RDW-SD group (> 42.5 fL, 12 cases), and the immune related indexes of the two groups were compared. Receiver operator characteristic curve (ROC) was drawn to evaluate the predictive value of RDW-SD on the severity of illness of coronavirus disease 2019 (COVID-19). RESULTS: Correlation analysis showed that RDW-SD was the only index related to PaO2 and PaCO2 on the first day of admission, which was negative correlation with PaO2 (r = -0.379, P = 0.047) and positive correlation with PaCO2 (r = 0.509, P = 0.006). The results of effects of different clinical characteristics on RDW-SD level showed that there was no statistically significant difference in RDW-SD between groups with different clinical characteristics (including male/female, ≥ 65 years old/< 65 years old, having/without hypertension, having/without diabetes, smoking/not smoking, having/without hyperpyrexia, with/without fever for 3 days, with/without respiratory symptoms, with/without digestive symptoms). It was suggested that RDW-SD be relatively stable and not affected by the patient's baseline level. The percentage of B cells in low RDW-SD group was higher than that in high RDW-SD group (23.01±3.01 vs. 15.34±5.34, P < 0.05), immunoglobulin G (IgG) level in low RDW-SD group was lower than that in high RDW-SD group (g/L: 11.43±3.20 vs. 15.42±1.54, P < 0.05). The area under ROC curve (AUC) of RDW-SD in evaluating severe cases was 0.83 [95% confidence interval (95%CI) was 0.59-1.06], which was close to multilobularinltration, hypo-lymphocytosis, bacterial coinfection, smoking history, hyper-tension and age (MuL BSTA score; AUC = 0.82, 95%CI was 0.51-1.12) and better than British Thoracic Society's modified pneumonia score (CURB-65 score; AUC = 0.70, 95%CI was 0.50-0.91). CONCLUSIONS: RDW-SD has significant evaluative effect on the severity of COVID-19 patients with Delta variants.

Study of the changes in immune indexes, pathogenic characteristics and related risk factors in children with viral diarrhea.

BACKGROUND: Infectious diarrhea is a serious hazard to children under 5 years old. The causative microorganisms are mostly viruses and bacteria, with different treatment required for each. Currently, early clinical differential diagnosis is difficult with the available testing methods. Therefore, new and more sensitive indicators of viral infection reflect the early stage of infection are needed. METHODS: We collected blood samples and fresh fecal samples from 100 children diagnosed with viral diarrhea who were treated in the outpatient clinic at Changzhou Tumor Hospital Affiliated to Soochow University from January 2018 to December 2020. The levels of interleukin-2 (IL-2), IL-6, tumor necrosis factor-α (TNF-α), and C-reactive protein (CRP) in serum, the erythrocyte sedimentation rate (ESR) and enterovirus were measured and compared with those in a matched healthy control group. Patients' demographic and risk factor data were collected by interviewing parents. RESULTS: The mean levels of IL-2, IL-6, TNF-α, ESR, and CRP in the viral diarrhea group were higher than those in the healthy control group. Except for IL-6, the differences in inflammatory factors between groups were statistically significant (P<0.05). Human rotavirus (HRV) infection was the highest. Children's hand-washing habits, weekly disinfection of toys, and vaccination were protective factors for viral diarrhea, and indoor flies were an important risk factor (P<0.05). CONCLUSIONS: The serum levels of markers were significantly increased in the children with viral diarrhea. HRV was the main pathogen, and mixed infections existed. Infection control requires hand washing, regular disinfection of toys, vaccination and preventing indoor flies.

An unusual elateroid lineage from mid-Cretaceous Burmese amber (Coleoptera: Elateroidea).

We here report a new elateroid, Anoeuma lawrencei Li, Kundrata and Cai gen. et sp. nov., from mid-Cretaceous Burmese amber. Though superficially similar to some soft-bodied archostematans, Anoeuma could be firmly placed in the polyphagan superfamily Elateroidea based on the hind wing venation. Detailed morphological comparisons between extant elateroids and the Cretaceous fossils suggest that the unique character combination does not fit with confidence into any existing soft-bodied elateroid group, although some characters indicate possible relationships between Anoeuma and Omalisinae. Our discovery of this new lineage further demonstrates the past diversity and morphological disparity of soft-bodied elateroids.

Novel role of LINC01013/miR-6795-5p/FMNL3 axis in the regulation of hepatocellular carcinoma stem cell features.

Cancer stem cells (CSCs) are major contributors to tumor initiation, recurrence, and metastasis of hepatocellular carcinoma (HCC). Some long non-coding RNAs have been reported as modulators of stem-like properties in cancer cells. However, the role of LINC01013 in liver CSCs has not yet been clarified. In this study, we aimed to elucidate the expression pattern and functions of LINC01013 in HCC. HCC tissues and normal controls were collected, and the expression pattern of LINC01013 and miR-6795-5p was identified by quick real-time polymerase chain reaction. Cell counting kit-8 assay, colony formation, and spheroid formation were performed to measure cell viability, proliferation, and self-renewal of HCC cell lines. The expression of stem markers was detected by western blot analysis. The effect of LINC01013 on viability, proliferation, and stem-like properties was detected through gain-of-function and loss-of-function experiments. The direct interaction among LINC01013, miR-6795-5p, and FMNL3 was testified by dual-luciferase reporter gene assay. Tumor-bearing mice were constructed to ascertain the functions of LINC01013 in vivo. HCC tissues showed increased LINC01013 and FMNL3 expression, while it showed a decreased miR-6795-5p expression as compared to the relative controls. Moreover, the high level of LINC01013 was closely related to the poor prognosis of HCC patients. LINC01013 directly binds to miR-6795-5p and subsequently relieves FMNL3. Silencing LINC01013, FMNL3, or overexpression of miR-6795-5p could suppress spheroid and colony formation, proliferation, as well as expression of stemness markers in HepG2 and SNU-182 cells. LINC01013 knockdown suppressed growth and stem-like traits of HCC cells in vivo by reducing FMNL3 expression. LINC01013/miR-6795-5p/FMNL3 axis may be a novel therapeutic target for HCC.

Effect of Walnut Meal Peptides on Hyperlipidemia and Hepatic Lipid Metabolism in Rats Fed a High-Fat Diet.

As a natural active substance that can effectively improve blood lipid balance in the body, hypolipidemic active peptides have attracted the attention of scholars. In this study, the effect of walnut meal peptides (WMP) on lipid metabolism was investigated in rats fed a high-fat diet (HFD). The experimental results show that feeding walnut meal peptides counteracted the high-fat diet-induced increase in body, liver and epididymal fat weight, and reduce the serum concentrations of total cholesterol, triglycerides, and LDL-cholesterol and hepatic cholesterol and triglyceride content. Walnut meal peptides also resulted in increased HDL-cholesterol while reducing the atherosclerosis index (AI). Additionally, the stained pathological sections of the liver showed that the walnut meal peptides reduced hepatic steatosis and damage caused by HFD. Furthermore, walnut meal peptide supplementation was associated with normalization of elevated apolipoprotein (Apo)-B and reduced Apo-A1 induced by the high-fat diet and with favorable changes in the expression of genes related to lipid metabolism (LCAT, CYP7A1, HMGR, FAS). The results indicate that walnut meal peptides can effectively prevent the harmful effects of a high-fat diet on body weight, lipid metabolism and liver fat content in rats, and provide, and provide a reference for the further development of walnut meal functional foods.

Pediatric blastic plasmacytoid dendritic cell neoplasm: report of four cases and review of literature.

Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare and aggressive hematological malignancy with poor outcome. Four children with BPDCN treated at our hospital were enrolled. All the four cases presented with cutaneous lesions. Bone marrow and central nervous system was involved in 50% and 25% of patients, respectively. The whole exome sequencing analysis revealed that KMT2 family genes were the most frequently mutated (4/4, 100%), followed by IKZF2 (2/4, 50%). The point mutation p.D348N was found in three patients and one patient had p.C394Y mutation in the KMT2C gene. Translocation of KMT2A-MLLT3 was found in Case 2. Case 1 had complex karyotype, who was induced by acute myeloid leukemia-like regimens. Although he received allogeneic hematopoietic stem cell transplantation twice as well as CD123 chimeric antigen receptor T cell therapy, the disease still progressed and he died 37 months after diagnosis. The other three patients were treated with Interfant-99 protocol. They tolerated the therapy well without significant toxicities and now in complete remission so far with a median follow up time of 9 months. More studies are needed to address the question whether the complex karyotype and KMT2 family genes are the causes of the relapse and refractory in BPDCN.

On the Nosodendridae from mid-Cretaceous amber of northern Myanmar (Coleoptera: Nosodendroidea).

Nosodendridae is a small polyphagan beetle family with a sparse fossil record. Herein, the fossil Nosodendridae from mid-Cretaceous Burmese amber (ca. 99 Ma) are systematically reviewed. Nosodendron cretaceum Deng et al. is transferred into Archaenosodendron Li Cai gen. nov., as A. cretaceum (Deng et al.) comb. nov., primarily based on the morphology of prosternum. Three new species of Archaenosodendron from Burmese amber, A. explanatum Li Cai sp. nov., A. remotidens Li Cai sp. nov., and A. angulare Li Cai sp. nov., are also described and illustrated. A key to nosodendrid genera and species from Burmese amber is provided.

Lymphatic blood filling in CLEC-2-deficient mouse models.

C-type lectin-like receptor 2 (CLEC-2) is considered as a potential drug target in settings of wound healing, inflammation, and infection. A potential barrier to this is evidence that CLEC-2 and its ligand podoplanin play a critical role in preventing lymphatic vessel blood filling in mice throughout life. In this study, this aspect of CLEC-2/podoplanin function is investigated in more detail using new and established mouse models of CLEC-2 and podoplanin deficiency, and models of acute and chronic vascular remodeling. We report that CLEC-2 expression on platelets is not required to maintain a barrier between the blood and lymphatic systems in unchallenged mice, post-development. However, under certain conditions of chronic vascular remodeling, such as during tumorigenesis, deficiency in CLEC-2 can lead to lymphatic vessel blood filling. These data provide a new understanding of the function of CLEC-2 in adult mice and confirm the essential nature of CLEC-2-driven platelet activation in vascular developmental programs. This work expands our understanding of how lymphatic blood filling is prevented by CLEC-2-dependent platelet function and provides a context for the development of safe targeting strategies for CLEC-2 and podoplanin.

Three new diterpenoids from Euphorbia peplus.

Three new diterpenoids (1-3) (two abietane type diterpenoids and a paralianone type diterpenoid), together with four known compounds (4-7) were isolated from the whole plants of Euphorbia peplus. Their structures were elucidated through spectroscopic analysis and physicochemical characteristics. The cytotoxic activities of compounds 1-7 against five human tumour cell lines were evaluated, however, they were inactive at the concentration of 40 µM. The compound 3 enhanced lysosomal biogenesis with Lyso Tracker staining intensity of 132.6%.