The m6A Reader YTHDF2 Promotes Bladder Cancer Progression by Suppressing RIG-I-Mediated Immune Response.

N6-Methyladenosine (m6A) is the most prevalent internal modification of mammalian mRNAs. Recent studies have shown that m6A methyltransferases METTL3 and METTL14 play important roles in urothelial bladder carcinoma (BLCA). To provide a more comprehensive understanding of the m6A regulatory landscape in bladder cancer, we investigated the role of YTHDF2, a crucial m6A reader, in BLCA. YTHDF2 was frequently upregulated at both the RNA and protein level in BLCA. Functionally, YTHDF2 promoted the proliferation and tumor growth of BLCA cells in vitro and in vivo, respectively. Integrative RNA sequencing and m6A sequencing analyses identified RIG-I as a downstream target of YTHDF2. Mechanistically, YTHDF2 bound to the coding sequence of DDX58 mRNA, which encodes RIG-I, and mediated its degradation in an m6A-dependent manner. Knockdown of RIG-I inhibited apoptosis and promoted the proliferation of BLCA cells. Depleting RIG-I was also able to reverse the effects of YTHDF2 deficiency. YTHDF2-deficient BLCA cells implanted orthotopically in recipient mice activated an innate immune response and promoted recruitment of CD8+ T lymphocytes into the tumor bed and the urothelium. Moreover, YTHDF2 deficiency enhanced the efficacy of Bacillus Calmette-Guérin immunotherapy treatment. This study reveals that YTHDF2 acts as an oncogene in BLCA. YTHDF2 inhibits RIG-I to facilitate immune evasion, supporting testing YTHDF2 inhibition in combination with immunotherapy. SIGNIFICANCE: YTHDF2 regulates RIG-I-mediated innate immune signaling to support bladder cancer progression, highlighting the functional importance of m6A modifications in bladder cancer and uncovering therapeutic opportunities to improve patient outcomes.

Nilaparvata lugens ERR2 regulates moulting and ovary development is related to hormone signalling.

The nuclear receptor (NR) superfamily is one of the largest groups of transcription factors in living organisms. Oestrogen related receptor (ERR) is a class of nuclear receptors closely related to oestrogen receptors (ERs). In this study, the Nilaparvata lugens (N. lugens) ERR2 (NlERR2) was cloned, and the expression of NlERR2 was detected by qRT-PCR to explore the distribution of NlERR2 during development and in different tissues. Using RNAi and qRT-PCR, the interaction between NlERR2 and related genes of the 20-hydroxyecdysone (20E) and juvenile hormone (JH) signalling pathways was studied. The results showed that topical application of 20E and juvenile hormone III (JHIII) affected the expression of NlERR2, and NlERR2 could affect the expression of genes related to 20E and JH signalling pathways. Furthermore, NlERR2 and JH/20E hormone signalling-related genes affect moulting and ovarian development. NlERR2 and NlE93/NlKr-h1 affect the transcriptional expression of Vg-related genes. In summary, NlERR2 is related to hormone signalling pathways, which is also related to the expression of Vg and Vg related genes. Brown planthopper is one of the most important rice pests. This study provides an important basis for mining new targets for pest control.

The Function of Nilaparvata lugens (Hemiptera: Delphacidae) E74 and Its Interaction With ßFtz-F1.

Drosophila E74 is an early gene located in the polytene chromosome 74EF puff position. E74 controls the production of late genes, indicating that it plays a crucial role in this cascade model. Nilaparvata lugens E74 is closely related to Diaphorina citri, Bemisia tabaci, and Laodelphax striatellus. After downregulating E74, molting, and nymphal mortality were increased, and ovarian development was delayed. Moreover, the expression of Vg was reduced at the transcriptional level, as measured by qRT-PCR, and the content of Vg protein was reduced, as detected by Western blotting. After downregulating E74, the expression of hormone-related genes, including Tai, ßFtz-F1, Met, Kr-h1, UspA, UspB, E93, and Br, was changed. The expression of E74 was significantly decreased after downregulating hormone-related genes. When the expression of E74 and ßFtz-F1 was downregulated together, nymph mortality and molting mortality were higher than those when E74 or ßFtz-F1 was downregulated alone. Thus, E74 probably interacts with ßFtz-F1 at the genetic level. In summary, this study showed that E74 plays a crucial role in the development, metamorphosis and reproduction of N. lugens, possibly via the interaction with ßFtz-F1 at the genetic level. This study provides a basis for the development of new target-based pesticides and new methods for the effective control of N. lugens.

Applicability of 14 Formulas for Estimating Glomerular Filtration Rate in the Evaluation of Renal Function before and after Nephron-Sparing Surgery in Patients with Renal Tumors.

To compare the applicability of 14 equations of estimating glomerular filtration rate (eGFR) before and after nephron-sparing surgery (NSS) for renal function assessment of patients with renal tumors. Preoperative and postoperative GFR is measured by emission computed tomography (ECT) with 99mTc-DTPA as an imaging agent as reference GFR (rGFR) to compare with all formulas. Spearman correlation analysis and Bland-Altman agreement analysis were used to evaluate the correlation between rGFR and eGFR1 to 14 before and after surgery. A total of 50 cases including 22 males and 28 females were included. The results of preoperative eGFR1-14 correlated with rGFR (P < 0.05). The calculation results of all estimation formulas have a significant correlation with preoperative GFR. Preoperative MDRD-I, CKD-EPI SCysC, and FAS Scr-SCysC have good consistency. The CG formula has the highest precision and FAS Scr-SCysC has the highest accuracy. A total of 30 patients followed up after surgery, and postoperative rGFR correlated with CG, CKD-EPI, FAS, and BIS formulas (P < 0.05). But postoperative rGFR has no significant correlation with MDRD and Schwartz (P > 0.05). Postoperative CKD-EPI Scr-SCysC has best consistency, and FAS Scr-SCysC has the highest accuracy and precision. Our data suggest that eGFR equations evaluated by both serum creatinine (Scr) and cystatin C (SCysC) is not necessarily better than those evaluated by one of them alone. Among all enrolled equations, FAS Scr-SCysC is the best one to evaluate postoperative GFR in patients with renal tumors.

Clinical Performance Study of a New Fully Automated Red Blood Cell Permeability Fragility Analyzer.

In order to verify the applicability of the erythrocyte fragility test (EFT) carried out by the new fully automatic erythrocyte permeability fragility analyzer RA-800 for thalassemia screening, a total of 100 cases of suspected thalassemia patients who underwent pregnancy examinations at Luohu District People's Hospital are included. The results of a new automatic erythrocyte permeability fragility analyzer RA-800 are compared with the results of the detection system composing of the KOFA erythrocyte fragility test kit currently used in clinical laboratories. The diagnosis confirmed by genetic testing is used as the gold standard to evaluate the applicability of RA-800. The sensitivity, specificity, and accuracy of the new automatic erythrocyte permeability fragility analyzer RA-800 screening for thalassemia were 66.67%, 92.86%, and 85.00%. The KOFA direct colorimetries are 76.67%, 81.43%, and 80.00%. The kappa value for the screening of thalassemia was 0.558, which concludes that the consistency was moderate. The ROC curve indicates that both two methods had diagnostic significance for the diagnostic value of thalassemia. The new automatic erythrocyte permeability fragility analyzer RA-800 is suitable for thalassemia screening, and the performance indexes meet the clinical requirements.

Role of Groucho and Groucho1-like in Regulating Metamorphosis and Ovary Development in Nilaparvata lugens (Stål).

Juvenile hormone and ecdysone are key regulators in the metamorphosis and development. Grocho (Gro) is a highly conserved protein required for metamorphosis and development. Brown planthopper (Nilaparvata lugens) is a major pest affecting rice production in China and many Asian countries. Although the molecular function of Gro has been investigated in holometabolous insects such as Aedes aegypti and Drosophila melanogaster, their role in the hemimetabolous insect, brown planthopper, and the relationship between NlGro/NlGro1-L and JH/ecdysone signaling pathway, remained unknown. In this study, NlGroucho (NlGro) and NlGroucho1-like (NlGro1-L) were cloned. An analysis of the predicted protein sequence showed that NlGro has highly conserved Q domain and WD40 domain, and NlGro1-L has a highly conserved WD40 domain. The expression profiles of both genes were studied by quantitative real-time PCR (qRT-PCR). Their relative expressions were high in egg, head, wing, ovary, and testis. NlGro and NlGro1-L were found to interact genetically with juvenile hormone and ecdysone signaling by hormone treatment and RNAi of JH/ecdysone signaling-related genes. Moreover, when NlGro or NlGro1-L was down-regulated alone, the survival rate was decreased, the ovarian development was delayed, and the oviposition was also affected. All defects were aggravated when NlGro and NlGro1-L were down-regulated together. This study will help to develop new pesticides on the basis of the function of NlGro and NlGro1-L, and provide new possibilities for the control of Nilaparvata lugens.

Multi-Omics Characterization of Tumor Microenvironment Heterogeneity and Immunotherapy Resistance Through Cell States-Based Subtyping in Bladder Cancer.

Due to the strong heterogeneity of bladder cancer (BC), there is often substantial variation in the prognosis and efficiency of immunotherapy among BC patients. For the precision treatment and assessment of prognosis, the subtyping of BC plays a critical role. Despite various subtyping methods proposed previously, most of them are based on a limited number of molecules, and none of them is developed on the basis of cell states. In this study, we construct a single-cell atlas by integrating single cell RNA-seq, RNA microarray, and bulk RNA-seq data to identify the absolute proportion of 22 different cell states in BC, including immune and nonimmune cell states derived from tumor tissues. To explore the heterogeneity of BC, BC was identified into four different subtypes in multiple cohorts using an improved consensus clustering algorithm based on cell states. Among the four subtypes, C1 had median prognosis and best overall response rate (ORR), which characterized an immunosuppressive tumor microenvironment. C2 was enriched in epithelial-mesenchymal transition/invasion, angiogenesis, immunosuppression, and immune exhaustion. Surely, C2 performed the worst in prognosis and ORR. C3 with worse ORR than C2 was enriched in angiogenesis and almost nonimmune exhaustion. Displaying an immune effective environment, C4 performed the best in prognosis and ORR. We found that patients with just an immunosuppressive environment are suitable for immunotherapy, but patients with an immunosuppressive environment accompanied by immune exhaustion or angiogenesis may resist immunotherapy. Furthermore, we conducted exploration into the heterogeneity of the transcriptome, mutational profiles, and somatic copy-number alterations in four subtypes, which could explain the significant differences related to cell states in prognosis and ORR. We also found that PD-1 in immune and tumor cells could both influence ORR in BC. The level of TGFß in a cell state can be opposite to the overall level in the tissues, and the level in a specific cell state could predict ORR more accurately. Thus, our work furthers the understanding of heterogeneity and immunotherapy resistance in BC, which is expected to assist clinical practice and serve as a supplement to the current subtyping method from a novel perspective of cell states.

Budget and fate of sedimentary trace metals in the Eastern China marginal seas.

To achieve a better understanding of global biogeochemical cycle and budget of trace metals (TM) in the world's ocean, a comprehensive study of the level, fate and burial flux of TM in surface sediment from the Eastern China Marginal Seas (ECMS) was conducted. The results indicated that Pb, Zn, Cu, and Cr were highly concentrated in mud deposits and primarily controlled by the natural processes of sediment composition and regional hydrodynamics, whereas As had a close association with Mn and was mainly derived from anthropogenic activity. The sediment mass inventories of Pb, Zn, Cu, Cr, and As in the ECMS (~220,780 km2) were estimated to be 28,324, 92,192, 23,434, 94,560 and 11,968 t/yr, respectively. A mass budget model revealed that riverine runoff, coastal erosion input and atmospheric deposition (dry and wet) constituted 62-76%, 15-37% and 2-9% of the total Pb, Zn and Cr influxes, respectively, while more than 4,690 t of Cu annually outflowed to the open seas to balance the budget. More importantly, we found that the sum of the estimates of these contributions tended to fall short of the calculated depositional fluxes of As, implying that anthropogenic activities probably have altered the natural geochemical cycle of As. Our results suggest that the ECMS constitutes a major final repository of TM at the Asia scale; however, the burial fluxes of trace metals are expected to decrease due to enhanced environmental investments by the Chinese government and decreased suspended particulate TM loads from the Chinese major rivers.

Predicting Errors, Violations, and Safety Participation Behavior at Nuclear Power Plants.

Commissioning workers at nuclear power plants have long been ignored in previous studies, although their performance is closely related to the overall safety of plants. This study aimed to explain and predict three types of behavior, i.e., errors, violations, and safety participation, of commissioning workers, under the general framework of the theory of planned behavior (TPB) and by considering organization and planning factors. The validity of the model was evaluated with a sample of 167 commissioning workers who completed a self-reported questionnaire. The results showed that perceived behavioral control, along with organization and planning, significantly affected all types of behavior. It was also found that violations and errors were a direct result of attitude. Besides, errors were predicted by subjective norm; unexpectedly, this occurred in a positive way. These findings revealed the underlying mechanisms for the development of errors, violations, and safety participation among commissioning workers and provided practical implications for safety improvement at the commissioning workplace.

[Electricity production from surplus sludge using microbial fuel cells].

A single-chamber and membrane-less microbial fuel cells were successfully started up using anaerobic sludge as inoculums without any chemical substance for 20 d. The electricity generation of the microbial fuel cell using surplus sludge as fuel and the change of substrate were investigated. The results showed that the obtained maximum voltage and power density were 495 mV and 44 mW x m(-2) (fixed 1,000 Omega), and the internal resistance was about 300 Omega during steady state. In a cycle, the removal efficiency of SS and VSS were 27.3% and 28.7%, pH was 6.5-8.0. In addition, the COD increased from 617 mg x L(-1) to 1,150 mg x L(-1) and decreased afterwards with time. The change of glucose was similar to that of COD, glucose increased from 47 mg x L(-1) to 60 mg x L(-1) and decreased afterwards with time. Consequently, the microbial fuel cell can transform chemical energy of surplus sludge into the cleanest electrical energy, and it provides a new way of sludge recycling.