CT whole lung radiomic nomogram: a potential biomarker for lung function evaluation and identification of COPD.

BACKGROUND: Computed tomography (CT) plays a great role in characterizing and quantifying changes in lung structure and function of chronic obstructive pulmonary disease (COPD). This study aimed to explore the performance of CT-based whole lung radiomic in discriminating COPD patients and non-COPD patients. METHODS: This retrospective study was performed on 2785 patients who underwent pulmonary function examination in 5 hospitals and were divided into non-COPD group and COPD group. The radiomic features of the whole lung volume were extracted. Least absolute shrinkage and selection operator (LASSO) logistic regression was applied for feature selection and radiomic signature construction. A radiomic nomogram was established by combining the radiomic score and clinical factors. Receiver operating characteristic (ROC) curve analysis and decision curve analysis (DCA) were used to evaluate the predictive performance of the radiomic nomogram in the training, internal validation, and independent external validation cohorts. RESULTS: Eighteen radiomic features were collected from the whole lung volume to construct a radiomic model. The area under the curve (AUC) of the radiomic model in the training, internal, and independent external validation cohorts were 0.888 [95% confidence interval (CI) 0.869-0.906], 0.874 (95%CI 0.844-0.904) and 0.846 (95%CI 0.822-0.870), respectively. All were higher than the clinical model (AUC were 0.732, 0.714, and 0.777, respectively, P < 0.001). DCA demonstrated that the nomogram constructed by combining radiomic score, age, sex, height, and smoking status was superior to the clinical factor model. CONCLUSIONS: The intuitive nomogram constructed by CT-based whole-lung radiomic has shown good performance and high accuracy in identifying COPD in this multicenter study.

MicroRNA-27a-3p targeting Vangl1 and Vangl2 inhibits cell proliferation in mouse granulosa cells.

BACKGROUND: Mammalian folliculogenesis is the complex process through which primordial follicles develop into preovulatory follicles. The chief function of ovarian follicle granulosa cells is to play a vital role in the growth, development and atresia of ovarian follicles via gap junctions. Increasing evidence suggests that microRNAs (miRNAs) are essential regulators of granulosa cell apoptosis or proliferation. METHODS: The expression level of miR-27a-3p, myogenic differentiation (MyoD), Vangl1 and Vangl2 was investigated by Real-time quantitative PCR (RT-qPCR) and Western blot. Luciferase reporter assay, bioinformatics analysis and ChIP-PCR was used to detect the binding sites between miR-27a-3p, transcription factor and target genes. KEGG pathway analyses were performed to reveal the predicted targets of miR-27a-3p. Ethynyl deoxyuridine (EdU) proliferation assay was used to measure cell proliferation. RESULTS: To explore the underlying mechanisms of the miR-27a-3p function in the development of mouse granulosa cells (mGCs), we screened for the target genes of miR-27a-3p, confirmed its interaction with Vangl1 and Vangl2 and elucidated their roles in mGCs. MiR-27a-3p inhibited the proliferation of mGCs, whereas target genes Vangl1 and Vangl2 had the opposite effect. In addition, the transcription factor MYOD bound to and activated the promoter of miR-27a-3p. MiR-27a-3p suppressed Vangl1 and Vangl2 expression by targeting their 3'-untranslated region (3'-UTR). Furthermore, Vangl1 and Vangl2 suppressed the Wnt pathway by reducing the expression of ß-catenin and B-cell lymphoma/leukemia-2 (Bcl-2). CONCLUSION: These findings indicate a pro-survival mechanism of the MyoD/miR-27a-3p/Vangl1/Vangl2 axis for granulosa cell proliferation and suggest a novel target for the improvement of female fertility.

Streptococcus mutans dexA affects exopolysaccharides production and biofilm homeostasis.

OBJECTIVES: The study aimed to evaluate the role of Streptococcus mutans (S. mutans) dexA gene on biofilm structure and microecological distribution in multispecies biofilms. MATERIALS AND METHODS: A multispecies biofilm model consisting of S. mutans and its dexA mutants, Streptococcus gordonii (S. gordonii) and Streptococcus sanguinis (S. sanguinis) was constructed, and bacterial growth, biofilm architecture and microbiota composition were determined to study the effect of the S. mutans dexA on multispecies biofilms. RESULTS: Our results showed that either deletion or overexpression of S. mutans dexA had no effect on the planktonic growth of bacterium, while S. mutans dominated in the multispecies biofilms to form cariogenic biofilms. Furthermore, we revealed that the SmudexA+ group showed structural abnormality in the form of more fractures and blank areas. The morphology of the SmudexA group was sparser and more porous, with reduced and less agglomerated exopolysaccharides scaffold. Interestingly, the microbiota composition analysis provided new insights that the inhibition of S. gordonii and S. sanguinis was alleviated in the SmudexA group compared to the significantly suppressed condition in the other groups. CONCLUSION: In conclusion, deletion of S. mutans dexA gene re-modules biofilm structure and microbiota composition, thereby leading to decreased cariogenicity. Thus, the S. mutans dexA may be an important target for regulating the cariogenicity of dental plaque biofilms, expecting to be a probiotic for caries control.

Sustainable treatment of nitrate-containing wastewater by an autotrophic hydrogen-oxidizing bacterium.

Bacteria are key denitrifiers in the reduction of nitrate (NO3 --N), which is a contaminant in wastewater treatment plants (WWTPs). They can also produce carbon dioxide (CO2) and nitrous oxide (N2O). In this study, the autotrophic hydrogen-oxidizing bacterium Rhodoblastus sp. TH20 was isolated for sustainable treatment of NO3 --N in wastewater. Efficient removal of NO3 --N and recovery of biomass nitrogen were achieved. Up to 99% of NO3 --N was removed without accumulation of nitrite and N2O, consuming CO2 of 3.25 mol for each mole of NO3 --N removed. The overall removal rate of NO3 --N reached 1.1 mg L-1 h-1 with a biomass content of approximately 0.71 g L-1 within 72 h. TH20 participated in NO3 --N assimilation and aerobic denitrification. Results from 15N-labeled-nitrate test indicated that removed NO3 --N was assimilated into organic nitrogen, showing an assimilation efficiency of 58%. Seventeen amino acids were detected, accounting for 43% of the biomass. Nitrogen loss through aerobic denitrification was only approximately 42% of total nitrogen. This study suggests that TH20 can be applied in WWTP facilities for water purification and production of valuable biomass to mitigate CO2 and N2O emissions.

A terahertz near-field nanoscopy revealing edge fringes with a fast and highly sensitive quantum-well photodetector.

We demonstrate the successful implementation of a terahertz (THz) quantum-well photodetector (QWP) for effective signal collection in a scattering-type scanning near-field optical microscope (s-SNOM) system. The light source is an electrically pumped THz quantum cascade laser (QCL) at 4.2 THz, which spectrally matches with the peak photoresponse of THz QWP. The sensitive THz QWP has a low noise equivalent power (NEP) of about 1.1 pW/Hz0.5 and a spectral response range from 2 to 7 THz. The fast-responding capability of the THz QWP is vital for detecting the rapidly tip-modulated THz light which can effectively suppress the background noise. The THz images of the nanostructure demonstrate a spatial resolution of about 95 nm, corresponding to ∼λ/752 at 4.2 THz. We experimentally investigate and theoretically interpret the formation of the fringes which appear at the edge position of a gold stripe in the THz near-field image.

The Impact of Consumer Purchase Behavior Changes on the Business Model Design of Consumer Services Companies Over the Course of COVID-19.

The COVID-19 pandemic has had a profound psychological and behavioral impact on people around the world. Consumer purchase behaviors have thus changed greatly, and consumer services companies need to adjust their business models to adapt to this change. From the perspective of consumer psychology, this paper explores the impact of consumer purchase behavior changes over the course of the pandemic on the business model design of consumer services companies using a representative survey of 1,742 individuals. Our results show that changes in consumer purchase behavior have a significant impact on the design of consumer services firms' business models. Specifically, changes in consumers' purchase object, motive, and timeframe are more likely to spark a novelty-centered business model design, whereas changes in purchase method tend to inspire an efficiency-centered one. Our findings provide a theoretical reference for consumer services companies in designing business models when faced with unexpected crises.

Small peptide targeting ANP32A as a novel strategy for acute myeloid leukemia therapy.

Clinic therapy of acute myeloid leukemia (AML) remains unsatisfactory that urges for development of novel strategies. Recent studies identified ANP32A as a novel biomarker of unfavorable outcome of leukemia, which promoted leukemogenesis by increasing H3 acetylation and the expression of lipid metabolism genes. It is of great significance to investigate whether targeting ANP32A is a novel strategy for leukemia therapy. To target ANP32A, we identified a peptide that competed with ANP32A to bind to histone 3 (termed as H3-binding peptide, H3BP). Disrupting ANP32A and H3 interaction by the overexpression of H3BP-GFP fusion protein mimicked the effect of ANP32A knockdown, impaired H3 acetylation on multiple locus of target genes, reduced proliferation, and caused apoptosis in leukemia cells. Furthermore, a synthesized membrane-penetrating peptide TAT-H3BP effectively entered into leukemia cells and phenocopied such effect. In vivo, TAT-H3BP showed potent efficacy against leukemia: Intra-tumor injection of TAT-H3BP significantly reduced the volume of subcutaneous tumors in nude mice and recipient mice engrafted with TAT-H3BP-pretreated 6133/MPL W515L cells exhibited ameliorated leukemia burden and prolonged survival. Noticeably, TAT-H3BP efficiently suppressed proliferation and colony-forming unit of human primary AML cells without affecting normal cord blood cells. Our findings demonstrate that intervening the physical interaction of ANP32A with H3 impairs the oncogenicity of ANP32A and may be a promising therapeutic strategy against AML.

Lipid Accumulation Product Combined With Urine Glucose Excretion Improves the Efficiency of Diabetes Screening in Chinese Adults.

Background: To compare the efficacy of lipid accumulation product (LAP) and urine glucose excretion (UGE) in predicting diabetes and evaluate whether the combination of LAP and UGE would help to improve the efficacy of using LAP alone or UGE alone in identifying diabetes. Methods: Data from 7485 individuals without prior history of diabetes who participated in a cross-sectional survey in Jiangsu, China, were analyzed. Each participant underwent an oral glucose-tolerance test. Operating characteristic curves (ROC) and logistic regression analyses were used to evaluate the performance of LAP and UGE in identification of newly diagnosed diabetes (NDM) and prediabetes (PDM). Results: For subjects with NDM, the area under the ROC curve was 0.72 for LAP and 0.85 for UGE, whereas for PDM, these values were 0.62 and 0.61, respectively. Furthermore, LAP exhibited a comparable sensitivity with UGE in detecting NDM (76.4% vs 76.2%, p = 0.31). In predicting PDM, LAP showed a higher sensitivity than UGE (66.4% vs 42.8%, p < 0.05). The combination of LAP and UGE demonstrated a significantly higher sensitivity than that of LAP alone and UGE alone for identification of NDM (93.6%) and PDM (80.1%). Moreover, individuals with both high LAP and high UGE had significantly increased risk of NDM and PDM than those with both low LAP and low UGE. Conclusions: The combination of LAP and UGE substantially improved the efficacy of using LAP and using UGE alone in detecting diabetes, and may be a novel approach for mass screening in the general population.

Traditional Chinese medicine symptom normalization approach leveraging hierarchical semantic information and text matching with attention mechanism.

Traditional Chinese medicine (TCM) symptom normalization is difficult because the challenges of the symptoms having different literal descriptions, one-to-many symptom descriptions and different symptoms sharing a similar literal description. We propose a novel two-step approach utilizing hierarchical semantic information that represents the functional characteristics of symptoms and develop a text matching model that integrates hierarchical semantic information with an attention mechanism to solve these problems. In this study, we constructed a symptom normalization dataset and a TCM normalization symptom dictionary containing normalization symptom words, and assigned symptoms into 24 classes of functional characteristics. First, we built a multi-label text classifier to isolate the hierarchical semantic information from each symptom description and count the corresponding normalization symptoms and filter the candidate set. Then we designed a text matching model of mixed multi-granularity language features with an attention mechanism that utilizes the hierarchical semantic information to calculate the matching score between the symptom description and the normalization symptom words. We compared our approach with other baselines on real-world data. Our approach gives the best performance with a Hit@ 1, 3, and 10 of 0.821, 0.953, and 0.993, respectively, and a MeanRank of 1.596, thus outperforming significantly regarding the symptom normalization task. We developed an approach for the TCM symptom normalization task and demonstrated its superior performance compared with other baselines, indicating the promise of this research direction.

ACOX1, regulated by C/EBPα and miR-25-3p, promotes bovine preadipocyte adipogenesis.

Acyl-coenzyme A oxidase 1 (ACOX1) is the first and rate-limiting enzyme in peroxisomal fatty acid ß-oxidation of fatty acids. Previous studies have reported that ACOX1 was correlated with the meat quality of livestock, while the role of ACOX1 in intramuscular adipogenesis of beef cattle and its transcriptional and post-transcriptional regulatory mechanisms remain unclear. In the present study, gain-of-function and loss-of-function assays demonstrated that ACOX1 positively regulated the adipogenesis of bovine intramuscular preadipocytes. The C/EBPα-binding sites in the bovine ACOX1 promoter region at -1142 to -1129 bp, -831 to -826 bp, and -303 to -298 bp were identified by promoter deletion analysis and site-directed mutagenesis. Electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation (ChIP) further showed that these three regions are C/EBPα-binding sites, both in vitro and in vivo, indicating that C/EBPα directly interacts with the bovine ACOX1 promoter and inhibits its transcription. Furthermore, the results from bioinformatics analysis, dual luciferase assay, site-directed mutagenesis, qRT-PCR, and Western blotting demonstrated that miR-25-3p directly targeted the ACOX1 3'UTR (3'UTR). Taken together, our findings suggest that ACOX1, regulated by transcription factor C/EBPα and miR-25-3p, promotes adipogenesis of bovine intramuscular preadipocytes via regulating peroxisomal fatty acid ß-oxidation.

Ecological and toxicological assessments of anthropogenic contaminants based on environmental metabolomics.

There has long been a great concern with growing anthropogenic contaminants and their ecological and toxicological effects on living organisms and the surrounding environment for decades. Metabolomics, a functional readout of cellular activity, can capture organismal responses to various contaminant-related stressors, acquiring direct signatures to illustrate the environmental behaviours of anthropogenic contaminants better. This review entails the application of metabolomics to profile metabolic responses of environmental organisms, e.g. animals (rodents, fish, crustacean and earthworms) and microorganisms (bacteria, yeast and microalgae) to different anthropogenic contaminants, including heavy metals, nanomaterials, pesticides, pharmaceutical and personal products, persistent organic pollutants, and assesses their ecotoxicological impacts with regard to literature published in the recent five years. Contaminant-induced metabolism alteration and up/down-regulation of metabolic pathways are revealed in typical organisms. The obtained insights of variations in global metabolism provide a distinct understanding of how anthropogenic contaminants exert influences on specific metabolic pathways on living organisms. Thus with a novel ecotechnique of environmental metabolomics, risk assessments of anthropogenic contaminants are profoundly demonstrated.

Characterization of XR_311113.2 as a MicroRNA Sponge for Pre-ovulatory Ovarian Follicles of Goats via Long Noncoding RNA Profile and Bioinformatics Analysis.

Long noncoding RNAs (lncRNAs) were identified recently as a large class of noncoding RNAs (ncRNAs) with a length ≥200 base pairs (bp). The function and mechanism of lncRNAs have been reported in a growing number of species and tissues. In contrast, the regulatory mechanism of lncRNAs in the goat reproductive system has rarely been reported. In the present study, we sequenced and analyzed the lncRNAs using bioinformatics to identify their expression profiles. As a result, 895 lncRNAs were predicted in the pre-ovulatory ovarian follicles of goats. Eighty-eight lncRNAs were differentially expressed in the Macheng black goat when compared with Boer goat. In addition, the lncRNA XR_311113.2 acted as a sponge of chi-miR-424-5p, as assessed via a luciferase activity assay. Taken together, our findings demonstrate that lncRNAs have potential effects in the ovarian follicles of goats and may represent a promising new research field to understand follicular development.

Characteristics of HIV-1 env genes from Chinese chronically infected donors with highly broad cross-neutralizing activity.

Knowledge about the special characteristics of HIV-1 envelope (env) glycoproteins in rare individuals developing >90% neutralization breadth in Chinese subtype B' slow progressors may provide insights for vaccine design against local viruses. We performed a cross-sectional analysis on 7 samples. We tested the neutralization breadth and geometric mean ID50 titers (GMTs) of these samples, and divided them into hBCN+ and hBCN- group according to whether their neutralization breadth >90%. We obtained env sequences in these samples through single genome amplification (SGA) assay. By comparing with hBCN-, subtype B chronically infected group (B-SP), and Chinese subtype B group (B-Database), we analyzed the characteristics of the env sequences of hBCN+ group. Longer V1 and V4 regions with more glycosylation sites were found in hBCN+ samples compared to hBCN- samples. Further analysis compared to B-SP and B-Database showed that hBCN+ group exhibited unique extra-long V1 region containing higher proportion of N-glycan sites and additional cysteines.

Highly efficient ammonium removal through nitrogen assimilation by a hydrogen-oxidizing bacterium, Ideonella sp. TH17.

Ideonella sp. TH17, an autotrophic hydrogen-oxidizing bacterium (HOB), was successfully enriched and isolated from activated sludge in a domestic wastewater treatment plant (WWTP). Batch experiments were conducted to identify the cell growth and ammonium (NH4+-N) removal, and to verify the pathways of nitrogen utilization under different conditions. At a representative NH4+-N concentration of 100 mg/L in domestic wastewater, it was the first time that a HOB strain achieved a nearly 100% ammonium removal. More than 90% of NH4+-N was assimilated to biomass nitrogen by strain TH17. Only a little of N2 (<10% of initial NH4+-N) was detected without N2O emission in aerobic denitrification process. Autotrophic NH4+-N assimilation contributed predominantly to biomass nitrogen production, supplemented by assimilatory nitrate (NO3--N) reduction under aerobic conditions. A total of 17 amino acids, accounting for 54.25 ± 1.98% of the dry biomass, were detected in the bacterial biomass harvested at 72 h. These results demonstrated that the newly isolated strain TH17 was capable of removing NH4+-N and recovering nutrients from wastewater efficiently. A new solution was thus provided by this HOB strain for ammonium treatment in sustainable WWTPs of future.

Skin transcriptome profiles associated with black- and white-coated regions in Boer and Macheng black crossbred goats.

To increase the current understanding of the gene-expression profiles in different skin regions associated with different coat colors and identify key genes for the regulation of color patterns in goats, we used the Illumina RNA-Seq method to compare the skin transcriptomes of the black- and white-coated regions containing hair follicles from the Boer and Macheng Black crossbred goat, which has a black head and a white body. Six cDNA libraries derived from skin samples of the white-coated region (n = 3) and black-coated region (n = 3) were constructed from three full-sib goats. On average, we obtained approximately 76.5 and 73.5 million reads for skin samples from black- and white-coated regions, respectively, of which 75.39% and 76.05% were covered in the genome database. A total of 165 differentially expressed genes (DEGs) were detected between these two color regions, among which 110 were upregulated and 55 were downregulated in the skin samples of white- vs. black-coated regions. The results of Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses revealed that some of these DEGs may play an important role in controlling the pigmentation of skin or hair follicles. We identified three key DEGs, i.e., Agouti, DCT, and TYRP1, in the pathway related to melanogenesis in the different skin regions of the crossbred goat. DCT and TYRP1 were downregulated and Agouti was upregulated in the skin of the white-coated region, suggesting a lack of mature melanocytes in this region and that Agouti might play a key developmental role in color-pattern formation. All data sets (Gene Expression Omnibus) are available via public repositories. In addition, MC1R was genotyped in 200 crossbred goats with a black head and neck. Loss-of-function mutations in MC1R as well as homozygosity for the mutant alleles were widely found in this population. The MC1R gene did not seem to play a major role in determining the black head and neck in our crossbred goats. Our study provides insights into the transcriptional regulation of two distinct coat colors, which might serve as a key resource for understanding coat color pigmentation in goats. The region-specific expression of Agouti may be associated with the distribution of pigments across the body in Boer and Macheng Black crossbred goats.

The N-terminal 1-55 residues domain of pyruvate dehydrogenase from Escherichia coli assembles as a dimer in solution.

The pyruvate dehydrogenase complex (PDHc) from Escherichia coli is a large protein complex consisting of multiple copies of the pyruvate dehydrogenase (E1ec), dihydrolipoamide acetyltransferase (E2ec) and dihydrolipoamide dehydrogenase (E3ec). The N-terminal domain (NTD, residues 1-55) of E1ec plays a critical role in the interaction between E1ec and E2ec and the whole PDHc activity. Using circular dichroism, size-exclusion chromatography and dynamic light scattering spectroscopy, we show that the NTD of E1ec presents dimeric assembly under physiological condition. Pull-down and isothermal titration calorimetry binding assays revealed that the E2ec peripheral subunit-binding domain (PSBD) forms a very stable complex with the NTD, indicating the isolated NTD functionally interacts with PSBD and the truncated E1ec (E1ec∆NTD) does not interact with PSBD. These findings are important to understand the mechanism of PDHc and other thiamine-based multi-component enzymes.

Uranium sequestration in sediment at an iron-rich contaminated site at Oak Ridge, Tennessee via. bioreduction followed by reoxidation.

This study evaluated uranium sequestration performance in iron-rich (30 g/kg) sediment via bioreduction followed by reoxidation. Field tests (1383 days) at Oak Ridge, Tennessee demonstrated that uranium contents in sediments increased after bioreduced sediments were re-exposed to nitrate and oxygen in contaminated groundwater. Bioreduction of contaminated sediments (1200 mg/kg U) with ethanol in microcosm reduced aqueous U from 0.37 to 0.023 mg/L. Aliquots of the bioreduced sediment were reoxidized with O2, H2O2, and NaNO3, respectively, over 285 days, resulting in aqueous U of 0.024, 1.58 and 14.4 mg/L at pH 6.30, 6.63 and 7.62, respectively. The source- and the three reoxidized sediments showed different desorption and adsorption behaviors of U, but all fit a Freundlich model. The adsorption capacities increased sharply at pH 4.5 to 5.5, plateaued at pH 5.5 to 7.0, then decreased sharply as pH increased from 7.0 to 8.0. The O2-reoxidized sediment retained a lower desorption efficiency at pH over 6.0. The NO3--reoxidized sediment exhibited higher adsorption capacity at pH 5.5 to 6.0. The pH-dependent adsorption onto Fe(III) oxides and formation of U coated particles and precipitates resulted in U sequestration, and bioreduction followed by reoxidation can enhance the U sequestration in sediment.

Mercury methylation by Geobacter metallireducens GS-15 in the presence of Skeletonema costatum.

In this study, bacterial mercury (Hg) methylation was investigated under the influence of red-tide algae of Skeletonema costatum (S. costatum). The distribution and speciation of total mercury (THg) and methylmercury (MeHg) were profiled by employing Geobacter metallireducens (G. metallireducens GS-15) as the methylating bacteria. G. metallireducens GS-15 showed different capabilities in methylating different inorganic forms of Hg(II) (HgCl2) and Hg(II)-Algae (HgCl2 captured by S. costatum) to MeHg. In the absence of S. costatum, a maximum methylation efficiency of 4.31 ±â€¯0.47% was achieved with Hg(II) of 1-100 µg L-1, while accelerated MeHg formation rate was detected at a higher initial Hg(II) concentration. In the presence of S. costatum, there were distinct changes in the distribution of THg and MeHg by altering the bioavailability of Hg(II) and Hg(II)-Algae. A larger proportion of THg tended to be retained by a higher algal biomass, resulting in decreased methylation efficiencies. The methylation efficiency of Hg(II) decreased from 3.01 ±â€¯0.10% to 1.01 ±â€¯0.01% with 10-mL and 250-mL algal media, and that of Hg(II)-Algae decreased from 0.83 ±â€¯0.13% to 0.22 ±â€¯0.01% with 10-mL and 250-mL Hg(II)-Algae media. Fourier transform infrared spectrometry, surface charge properties and elemental compositions of S. costatum were used to infer that amine, carboxyl and sulfonate functional groups were most likely to interact with Hg(II) through complexation and/or electrostatic attraction. These results suggest that red-tide algae may be an influencing factor on bacterial Hg methylation in eutrophic water bodies.

The Inhibitory Effect of Celangulin V on the ATP Hydrolytic Activity of the Complex of V-ATPase Subunits A and B in the Midgut of Mythimna separata.

Celangulin V (CV) is a compound isolated from Celastrus angulatus Max that has a toxic activity against agricultural insect pests. CV can bind to subunits a, H, and B of the vacuolar ATPase (V-ATPase) in the midgut epithelial cells of insects. However, the mechanism of action of CV is still unclear. In this study, the soluble complex of the V-ATPase A subunit mutant TSCA which avoids the feedback inhibition by the hydrolysate ADP and V-ATPase B subunit were obtained and then purified using affinity chromatography. The H⁺K⁺-ATPase activity of the complex and the inhibitory activity of CV on ATP hydrolysis were determined. The results suggest that CV inhibits the ATP hydrolysis, resulting in an insecticidal effect. Additionally, the homology modeling of the AB complex and molecular docking results indicate that CV can competitively bind to the AB complex at the ATP binding site, which inhibits ATP hydrolysis. These findings suggest that the AB subunits complex is one of the potential targets for CV and is important for understanding the mechanism of interaction between CV and V-ATPase.

NFAT5 is Regulated by p53/miR-27a Signal Axis and Promotes Mouse Ovarian Granulosa Cells Proliferation.

MicroRNAs (miRNAs) play key roles in mammalian folliculogenesis (a complex process in which primordial follicles develop into mature oocytes) by inhibiting mRNA translation or by inducing its degradation, while the role of miRNA in folliculogenesis and regulation mechanism remain unclear. In this study, we explored the role of the p53/miR-27a/nuclear factor of activated T-cells 5 (NFAT5) signaling axis in mouse ovarian granulosa cell proliferation. Luciferase reporter assay, overexpression, site-directed mutagenesis, and chromatin immunoprecipitation (ChIP) assay results showed that the transcription factor p53 significantly decreased the expression level of miR-27a by binding to sites 4 (-646 to -637 bp) and 10 (-50 to -41 bp) of the miR-27a promoter. Moreover, miR-27a directly targeted the 3'-untranslated region of the target gene, NFAT5, to regulate its expression levels. p53 also upregulated the expression of NFAT5. Meanwhile, overexpression of NFAT5 strongly upregulated the mRNA and protein levels of the Wnt signaling genes, ß-catenin and B-Cell CLL/Lymphoma 2 (Bcl-2). In addition, NFAT5 promoted mouse granulosa cell proliferation; this was confirmed by EdU/Hoechst immunostaining. Taken together, our findings define a novel pathway p53/miR-27a/NFAT5, and NFAT5 regulates mouse granulosa cell functions via activating Wnt signaling pathway.