LMD and LC-MS-based chemical constituents and pharmacological effects assessment for two different processing methods of the root of Paeonia lactiflora Pall.

The plant of Paeonia lactiflora Pall. belongs to Ranunculaceae, and its root can be divided into two categories according to different processing methods, which included that one was directly dried without peeling the root of the P. lactiflora (PR), and the other was peeled the root of the P. lactiflora (PPR) after boiled and dried. To evaluate the difference of chemical components, UPLC-ESI-Q-Exactive Focus-MS/MS and UPLC-QQQ-MS were applied. The distribution of chemical components in different tissues was located by laser microdissection (LMD), especially the different ingredients. A total of 86 compounds were identified from PR and PPR. Four kind of tissues were isolated from the fresh root of the P. lactiflora (FPR), and 54 compounds were identified. Especially the content of gallic acid, albiflorin, and paeoniflorin with high biological activities were the highest in the cork, but they were lower in PR than that in PPR, which probably related to the process. To illustrate the difference in pharmacological effects of PR and PPR, the tonifying blood and analgesic effects on mice were investigated, and it was found that the tonifying blood and analgesic effects of PPR was superior to that of PR, even though PR had more constituents. The material basis for tonifying blood and analgesic effect of the root of P. lactiflora is likely to be associated with an increase in constituents such as paeoniflorin and paeoniflorin lactone after boiled and peeled. The study was likely to provide some theoretical support for the standard and clinical application.

Novel Perspectives in Chronic Kidney Disease-Specific Cardiovascular Disease.

Chronic kidney disease (CKD) affects > 10% of the global adult population and significantly increases the risk of cardiovascular disease (CVD), which remains the leading cause of death in this population. The development and progression of CVD-compared to the general population-is premature and accelerated, manifesting as coronary artery disease, heart failure, arrhythmias, and sudden cardiac death. CKD and CV disease combine to cause multimorbid cardiorenal syndrome (CRS) due to contributions from shared risk factors, including systolic hypertension, diabetes mellitus, obesity, and dyslipidemia. Additional neurohormonal activation, innate immunity, and inflammation contribute to progressive cardiac and renal deterioration, reflecting the strong bidirectional interaction between these organ systems. A shared molecular pathophysiology-including inflammation, oxidative stress, senescence, and hemodynamic fluctuations characterise all types of CRS. This review highlights the evolving paradigm and recent advances in our understanding of the molecular biology of CRS, outlining the potential for disease-specific therapies and biomarker disease detection.

Identification and validation of cuproptosis-related genes for diagnosis and therapy in nonalcoholic fatty liver disease.

In recent years, nonalcoholic fatty liver disease (NAFLD) has become a more serious public health issue worldwide. This study strived to investigate the molecular mechanism of pathogenesis of NAFLD and explore promising diagnostic and therapeutic targets for NAFLD. Raw data from GSE130970 were downloaded from the Gene Expression Omnibus database. We used the dataset to analyze the expression levels of cuproptosis-related genes in NAFLD patients and healthy controls to identify the differentially expressed cuproptosis-related genes (DECRGs). The relationship and potential mechanism between DECRGs and clinicopathological factors were examined by enrichment analysis and two consensus clustering methods. We screened key DECRGs based on Random Forest (RF), and then verified the key DECRGs in NAFLD patients, high-fat diet (HFD)-fed mice, and palmitic acid-induced AML12 cells. ROC analysis showed good diagnostic function of DECRGs in normal and NAFLD liver tissue. Two consensus clusters indicated the important role of cuproptosis in the development of NAFLD. We screened for key DECRGs (DLD, DLAT) based on RF and found a close relationship between the DECRGs and clinicopathological factors. We collected clinical blood samples to verify the differences in gene expression levels by qPCR. In addition, we further verified the expression levels of DLD and DLAT in HFD mice and AML12 cells, which showed the same results. This study provides a novel perspective on the pathogenesis of NAFLD. We identified two cuproptosis-related genes that are closely related to NAFLD. These genes may play a significant role in the molecular pathogenesis of NAFLD, which may be useful to make progress in the diagnosis and treatment of NAFLD.

CD276 promotes epithelial-mesenchymal transition in esophageal squamous cell carcinoma through the TGF-ß/SMAD signaling.

OBJECTIVE: Aberrant expression of CD276 has been reported in malignant tumors. However, the exact role and mechanisms of CD276 influence the progression of esophageal squamous cell carcinoma (ESCC) still need to be understood. METHODS: Bioinformatics analysis of data from The Cancer Genome Atlas and Gene Expression Omnibus databases, along with immunohistochemistry staining, was used to explore the expression patterns of CD276 in ESCC. Cell counting kit-8 and Transwell assays were employed to evaluate the effects of CD276 expression on tumor cell proliferation and motility. Western blotting and Transwell assays were used to explore the potential pathways through which CD276 mediates the progression of ESCC. Moreover, the in vivo role of CD276 in tumor progression was investigated by establishing a lung metastasis mouse model. RESULTS: A significant upregulation of CD276 was observed in ESCC tissues compared to adjacent tissues. The inhibition of CD276 had no evident impact on ESCC cell proliferation but notably hindered their migratory and invasive properties and the expression of epithelial-mesenchymal transition (EMT) markers. Inversely, overexpressing CD276 led to an upregulation of EMT markers, underscoring the capacity of CD276 to amplify the motility of ESCC cells. Furthermore, CD276 was found to enhance the migratory and invasive abilities of ESCC cells by activating the TGF-ß/SMAD signaling but not the PI3K/AKT pathway. In vivo studies demonstrated that CD276 facilitates pulmonary metastasis. CONCLUSION: CD276 is significant upregulation in ESCC tissues and facilitates the EMT process in ESCC cells via the TGF-ß/SMAD signaling, thus promoting the progression of ESCC.

Establishment and validation of a long-term prognosis prediction model for patients with non-small cell lung cancer.

Background: The incidence of non-small cell lung cancer ranks second among malignant tumors, while the mortality rate ranks first. We established a prediction model for the long-term prognosis of lung cancer patients to accurately identify patients with a high risk of postoperative death and provide a theoretical basis for improving the prognosis of patients with non-small cell lung cancer. Methods: The data of 277 non-small cell lung cancer patients who underwent radical lung cancer resection at Shanghai Fengxian District Central Hospital between January 2016 and December 2017 were retrospectively collected. The patients, who were followed up for 5 years, were divided into a deceased group (n=127) and survival group (n=150) according to whether the patients had died 5 years after surgery or not. The clinical characteristics of the two groups were observed, and the risk factors of death within 5 years of surgery in lung cancer patients were analyzed. A nomogram predictive model was then established to analyze the value of the model in predicting the death within 5 years of surgery in patients with non-small cell lung cancer. Results: Multivariate logistics regression analysis showed that carcinoembryonic antigen (CEA) >193.5 ng/mL, stage III lung cancer, peritumor invasion, and vascular tumor thrombus were independent risk factors of tumor-specific death after surgery in patients with non-small cell lung cancer (P<0.05). R 4.0.3 statistical software was used to randomly divide the dataset into a training set and validation set. The sample size of the training set was 194, and the sample size of the validation set was 83. The area under the receiver operating characteristic (ROC) curve was 0.850 [95% confidence interval (CI): 0.796-0.905] in the training set, and it was 0.779 (95% CI: 0.678-0.880) in the validation set. In the validation set, the model was assessed using the Hosmer-Lemeshow goodness-of-fit test, with a chi-square value of 9.270 and a P value of 0.320. Conclusions: Our model could accurately identify high risk of death within 5 years of surgery in non-small cell lung cancer patients. Strengthening the management of high-risk patients may help improve the prognosis of these patients.

Multi-dimensional safety risk assessment on coal mines under the profitability dilemma.

China is a major coal producer, with huge differences in coal production and safety situations between the South and the North. Taking province A as an example, its coal enterprises have low output, poor efficiency, backward equipment, and low-quality personnel. The output accounts for 0.08% of the country, and the number of deaths accounts for 2.2% of the country, the safety situation of coal enterprises in province A is severe. In order to study the safety risk situation of coal mines under difficult conditions, this paper screens 98 factor indexes including multiple subjects such as enterprise managers, front-line workers, government supervisors, external environment, work quality, stress factors, economic factors, and other dimensions. For different data, the indicator weights were calculated using triangular fuzzy number, entropy weight method, CRITIC method, and three rough set methods in a total of six methods. The comprehensive weights of the indicators were obtained using the proposed new comprehensive weight method. The current situation of safety work of four coal mining enterprises and three levels of government supervision departments was evaluated, and the evaluation results were compared with other existing data to verify the reliability of the evaluation system. The horizontal comparison of the evaluation results indicates the main problems of each subject; the longitudinal comparison points out the problems that need to be solved with the assistance of higher-level enterprises and the central government, and corresponding suggestions for coal mining enterprises and government departments are put forward to reduce the safety risks of troubled coal mining enterprises.

Experimental research on atomization process and dust reduction performance of swirl pressure nozzle.

In this study, experimental studies on atomization process and dust reduction performance of four swirl nozzles with different inlet/outlet diameter ratio (D) were performed. The results of the atomization process study of the nozzle show that with the increase of D, the droplet breakup range of the spray field is gradually increasing, but the droplet breakup intensity of the spray field is gradually decreasing. At D = 3.33 and 3.63, droplet breakup occurs mainly in the range of 0-4 mm in the strong turbulent region. At D = 3.75, droplet breakup occurs mainly in the range of 0-2 mm in the strong turbulent region. At D = 3.96, droplet breakup occurs mainly in the range of 0-1 mm in the strong turbulent region. Droplet breakup in the spray field at D = 3.33 and D = 3.67 was better than that at D = 3.75 and D = 3.96. From the dust reduction experimental results, the dust reduction efficiency increases and then decreases with the increase of D. The dust reduction efficiency is highest among the four nozzles at D = 3.67. Based on the dust reduction curves of four different D of nozzles, it is predicted that the optimal dust reduction condition will be achieved at D of 3.60, which provides a reference for the design and optimization of nozzles.

Research on the wetting mechanism of coal dust by different surfactants: combination of experimental characterization and molecular dynamics simulation.

Surfactants can reduce the surface tension of water and improve the efficiency of spray dust reduction, but the synergistic mechanism of composite surfactant solutions wetting coal dust remains unclear. In this study, sodium dodecyl sulfonate (SDDS)/sodium dodecylbenzene sulfonate (SDBS) solution and SDDS/primary alcohol ethoxylate (AEO-9) solution were prepared to wet three types of coal with different deterioration degrees. The surface tension, contact angle, and functional group composition were measured. The results show that SDDS/AEO-9 solution had lower surface tension and critical micelle concentration than SDDS/SDBS solution at the same mixing ratio. When the ratio of SDDS: SDBS was 2:1, it had the best wetting effect on coal dust. It is found that for SDDS/SDBS solutions, aliphatic hydrocarbons and surface tension have a positive correlation with the contact angle, and hydroxyl groups have a negative correlation with the contact angle. For SDDS/AEO-9 solution, only the surface tension and contact angle show a significant linear relationship. The main factors that affect the wetting change with the combination of surfactants. AEO-9 has an electrostatic shielding effect on SDDS, while SDBS increases the electrostatic repulsion between SDDS solution and electronegative functional groups of coal. The synergism among these surfactants plays an important role in the process of wetting coal.

Preparation and characterization of modified dual network dust suppression gel based on sodium alginate and soluble starch.

Aiming at the problems of complex environment and serious dust pollution in large open-pit coal yards, a dust suppression gel with a dual network structure was prepared by modifying the soluble starch and sodium alginate with iron ions. The changes of functional groups, thermal stability, and morphology structure before and after the reaction were analyzed by FTIR, TG-DSC, and SEM, and the formation mechanism of the dual network was revealed by XPS. Furthermore, the water absorption and water retention experiments proved that the dual network structure is more conducive to water retention than the single-layer network. According to molecular dynamics simulations and contact angle experiments, gel and adsorbed water molecules can approach coal dust molecules on their own to contact, wet, and combine with coal dust. The adhesion test proved that the dust suppression gel with iron ions had better adhesion to dust. The anti-freezing test shows that the dust suppression gel has good anti-freezing performance. The antifreeze test shows that the dust suppression gel still has excellent freeze-thaw resistance at the test temperature of -20℃. The mechanical property test shows that the dust suppressant gel can prevent the product from being damaged by external force. The acid and alkali resistance experiments showed that the acid and alkali resistance of the gel was improved under the condition of iron ion modification, and the flying of coal powder was effectively prevented. This research provides a new theoretical idea for coal dust control in complex environment.

Mechanical Properties and Tensile Model of Hemp-Fiber-Reinforced Poly(butylene adipate-co-terephthalate) Composite.

The preparation of a high-strength biodegradable plastic has always been the focus of academia. Here, we prepared two biodegradable composites using silane coupling-agent-modified hemp fibers (Si-HF) and unmodified hemp fibers (HF) with butylene adipate-co-terephthalate (PBAT), respectively. We compared the differences of Si-HF/PBAT and HF/PBAT in terms of micromorphology, density, mechanical properties, thermal stability and biodegradability. The Si-HF has better interface interaction between the hemp and the PBAT matrix than the HF, which makes Si-HF/PBAT have better tensile properties. Moreover, Si-HF/PBAT has stronger tensile strength and modulus than HF/PBAT. Our results also show that the two composites have good biodegradability. This study provides an important reference for the subsequent development and utilization of hemp fibers.

A covalent organic framework as a photocatalyst for window ledge cross-dehydrogenative coupling reactions.

A benzothiadiazole-involving donor-acceptor (D-A) covalent organic framework (COF), which has high crystallinity and strong light-harvesting capability (ranging from 300 to 800 nm), can serve as a highly effective photocatalyst for window ledge aerobic cross-dehydrogenative coupling (CDC) reactions (such as Mannich and aza-Henry reactions) even at a gram level.

High nitrogen in maize enriches gut microbiota conferring insecticide tolerance in lepidopteran pest Spodoptera litura.

Abuse of chemical fertilizers and insecticides has created many environmental and human health hazards. We hypothesized that high nitrogen (N) in crops changes insect gut microbiota leading to enhanced insecticide tolerance. We investigated the effect of high N in maize on gut microbiota and insecticide tolerance of the polyphagous pest Spodoptera litura. Bioassays showed that high N applied in both maize plants and artificial diets significantly enhanced larval growth but reduced larval sensitivity to the insecticide methomyl. High N promoted the gut bacterial abundance in the genus Enterococcus. Inoculation with two strains (E. mundtii and E. casseliflavus) isolated from the larval guts increased larval tolerance to methomyl. Incorporation of antibiotics in a high-N diet increased the larval sensitivity to methomyl. These findings suggest that excessive application of N fertilizer to crops can increase insecticide tolerance of insect pests via changing gut microbiota, leading to increased use of insecticides worldwide.

Quantitative analysis of size and regional distribution of corpora amylacea in the hippocampal formation of obstructive sleep apnoea patients.

Corpora amylacea (CoA) are spherical aggregates of glucose polymers and proteins within the periventricular, perivascular and subpial regions of the cerebral cortex and the hippocampal cornu ammonis (CA) subfields. The present study quantified the distribution of CoA in autopsied hippocampi of patients with obstructive sleep apnoea (OSA) using ethanolamine-induced fluorescence. CoA were observed in 29 of 30 patients (96.7%). They were most abundant in periventricular regions (wall of lateral ventricle, alveus, fimbria and CA4), rarely found in the CA3 and CA1, and undetectable in the CA2 or subiculum. A spatiotemporal sequence of CoA deposition was postulated, beginning in the fimbria and progressively spreading around the subpial layer until they extended medially to the wall of the lateral ventricle and laterally to the collateral sulcus. This ranked CoA sequence was positively correlated with CoA packing density (count and area fraction) and negatively correlated with CoA minimum diameters (p < 0.05). Although this sequence was not correlated with age or body mass index (BMI), age was positively correlated with the mean and maximum diameters of CoA. These findings support the view that the spatiotemporal sequence of CoA deposition is independent of age, and that CoA become larger due to the accretion of new material over time.

Novel Self-Calibrating Amperometric and Ratiometric Electrochemical Nanotip Microsensor for pH Measurement in Rat Brain.

Brain pH has been proven to be a key factor in maintaining normal brain function. The relationship between local pH fluctuation and brain disease has not been extensively studied due to lack of the accurate in situ analysis technology. Herein, we have for the first time proposed a voltammetric pH sensor by measuring the ratio of current signals instead of the previously reported potential based on the Nernst equation. Single-walled carbon nanotubes (CNT) were first self-assembled on the electrode surface of a carbon-fiber nanotip electrode (CFNE). Then, poly-o-phenylenediamine (PoPD) molecules were deposited as pH-responsive molecules through in situ electrochemical polymerization. The compact CFNE/CNT/PoPD exhibited a good redox process with the on-off-on ratiometric electrochemical response to pH ranging from 4.5 to 8.2, providing self-correction for in situ pH detection. Thus, the proposed sensor enabled the accurate measurement of pH with excellent selectivity even in the presence of proteins or electroactive species. In addition, the sensor showed high repeatability, reproducibility, and reversibility in measuring pH and even demonstrated good stability when it was exposed to air for 5 months. Finally, we successfully detected the fluctuation of pH in rat brains with cerebral ischemia and rat whole blood. Overall, this research not only provides a good tool for the detection of rat brain pH but also provides a new strategy for further designing nanosensors for intracellular or subcellular pH.

Synthesis and characteristics of a novel dust suppressant with good weatherability for controlling dust in open coal yards.

This study aims to synthesize a dust suppressant for controlling coal dust pollution in open yards using natural polymers. Guided by graft copolymerization theory, potassium persulfate acts as an initiator to excite the free radicals of collagen and sodium alginate, allowing them to combine with acrylic acid and acrylamide to form a new polymer. The TG curve indicates that the thermal stability of the polymer is superior to that of the raw material. Scanning electron microscope (SEM) images show that the product can reduce the generation of dust by bonding the pulverized coal. With CCD method, when the monomer concentration increases in a certain range, the cohesive ability of the product to coal increases first and then decreases. Initiators and crosslinkers showed the same pattern. A series of performance experiments show that the product has a dust suppression rate of 98.7% at a wind speed of 14 m/s, and maintain one of 94.5% at a wind speed of 8 m/s after a rainstorm. In addition, there was no significant loss in dust suppression performance and compressive strength of the solidified layer after wind and rain. Sunlight, low temperature, and high temperature have little influence on the dust suppression effect of the product, which indicates that the product has better weather resistance and helps to suppress dust for a longer time in open air conditions.

RNA-Seq Analyses of Midgut and Fat Body Tissues Reveal the Molecular Mechanism Underlying Spodoptera litura Resistance to Tomatine.

Plants produce secondary metabolites to provide chemical defense against herbivorous insects, whereas insects can induce the expression of detoxification metabolism-related unigenes in counter defense to plant xenobiotics. Tomatine is an important secondary metabolite in tomato (Lycopersicon esculentum L.) that can protect the plant from bacteria and insects. However, the mechanism underlying the adaptation of Spodoptera litura, a major tomato pest, to tomatine in tomato is largely unclear. In this study, we first found that the levels of tomatine in tomatoes subjected to S. litura treatment were significantly increased. Second, we confirmed the inhibitory effect of tomatine on S. litura by adding moderate amounts of commercial tomatine to an artificial diet. Then, we utilized RNA-Seq to compare the differentially expressed genes (DEGs) in the midgut and fat body tissues of S. litura exposed to an artificial diet supplemented with tomatine. In total, upon exposure to tomatine, 134 and 666 genes were upregulated in the S. litura midgut and fat body, respectively. These DEGs comprise a significant number of detoxification-related genes, including 7 P450 family genes, 8 glutathione S-transferases (GSTs) genes, 6 ABC transport enzyme genes, 9 UDP-glucosyltransferases genes and 3 carboxylesterases genes. Moreover, KEGG analysis demonstrated that the upregulated genes were enriched in xenobiotic metabolism by cytochrome P450s, ABC transporters and drug metabolism by other enzymes. Furthermore, as numerous GSTs were induced by tomatine in S. litura, we chose one gene, namely GSTS1, to confirm the detoxification function on tomatine. Expression profiling revealed that GSTS1 transcripts were mainly expressed in larvae, and the levels were the highest in the midgut. Finally, when larvae were injected with double-stranded RNA specific to GSTS1, the transcript levels in the midgut and fat body decreased, and the negative effect of the plant xenobiotic tomatine on larval growth was magnified. These results preliminarily clarified the molecular mechanism underlying the resistance of S. litura to tomatine, establishing a foundation for subsequent pest control.

Identification of a cytochrome P450 CYP6AB60 gene associated with tolerance to multi-plant allelochemicals from a polyphagous caterpillar tobacco cutworm (Spodoptera litura).

Generalist phytophagous insects adapt to adventurous chemical environment in a wide variety of host plants by extraordinary detoxifying metabolic abilities. However, how polyphagous insect cope with the diversity of plant defenses remains largely unknown and only a few counter-defense genes detoxifying a wide range of toxic secondary metabolites have been well characterized. Here, we identify a cytochrome P450 gene (CYP6AB60) from tobacco cutworm (Spodoptera litura) in response to three different plant's defense metabolites. After being exposed to artificial diet supplemented with coumarin (COU), xanthotoxin (XAN) or tomatine (TOM), activities of P450 and CYP6AB60 transcript levels in both midgut and fat body tissues were significantly increased. Developmental expression analysis revealed that CYP6AB60 was expressed highly during the larval stages, and tissue distribution analysis showed that CYP6AB60 was expressed extremely high in the midgut, which correspond to the physiological role of CYP6AB60 from S. litura larvae in response to plant allelochemicals. Furthermore, when larvae are injected with double-stranded RNA (dsRNA) specific to CYP6AB60, levels of this transcript in the midgut and fatbody decrease and the negative effect of plant's defense metabolites on larval growth is magnified. These data demonstrate that the generalist insect S. litura might take advantage of an individual detoxificative gene CYP6AB60 to toxic secondary metabolites from different host plants. The CYP6AB60 can be a potential gene to carry out RNAi-mediated crop protection against the major polyphagous pest S. litura in the future.

Exposure to Herbicides Prime P450-Mediated Detoxification of Helicoverpa armigera against Insecticide and Fungal Toxin.

With the long-term and large-scale use, herbicides have been well known to influence tritrophic interactions, particularly natural enemies of pests in agro-ecosystems. On the other hand, herbivorous insects, especially the generalist pests, have developed antagonistic interaction to different insecticides, toxic plant secondary metabolites, and even heavy metals. However, whether exposure to herbicides would affect resistance of insects against insecticides is largely unknown, especially in agricultural pests. Here, we first reported that pre-exposure to two widely used herbicides butachlor and haloxyfop-methyl for 48 h can prime the resistance of a generalist agricultural pest Helicoverpa armigera Hübner against insecticide methomyl and fungal toxin aflatoxin B1. In addition, there were no significant differences between control and herbicides-treated caterpillars on weight gain, pupal weight, and pupation rates, suggesting that exposure to herbicides induces resistance of H. armigera accompanied with no fitness cost. Moreover, by determining detoxifying enzyme activities and toxicity bioassay with additional inhibitor of cytochrome P450 piperonyl butoxide (PBO), we showed that exposure to herbicides might prime P450-mediated detoxification of H. armigera against insecticide. Based on these results, we propose that exposure to herbicides prime resistance of H. armigera against insecticide and fungal toxin by eliciting a clear elevation of predominantly P450 monooxygenase activities in the midgut and fat body.

Identification of Energy Metabolism Changes in Diabetic Cardiomyopathy Rats Using a Metabonomic Approach.

BACKGROUND/AIMS: Diabetic cardiomyopathy (DCM) is a serious complication of diabetes. It is therefore crucial to elucidate the characteristic metabolic changes that occur during the development of diabetes to gain an understanding the pathogenesis of this disease and identify potential drug targets involved. METHODS: 1H nuclear magnetic resonance-based metabonomics combined with HPLC measurements were used to determine the metabolic changes in isolated cardiac tissues after 5 weeks, 9 weeks, and 15 weeks in rats treated with streptozotocin. RESULTS: Pattern recognition analysis clearly discriminated the diabetic rats from time-matched control rats, suggesting that the metabolic profile of the diabetic group was markedly different from that of the controls. Quantitative analysis showed that the levels of energy metabolites, such as the high-energy phosphate pool (ATP and creatine), significantly decreased in a time-dependent manner. Correlation analysis revealed the inhibition of glycolysis and the tricarboxylic acid (TCA) cycle, enhanced lipid metabolism, and changes in some amino acids, which may have led to the decline in energy production in the DCM rats. CONCLUSIONS: The results indicated that the administration of energy substances or the manipulation of myocardial energy synthesis induced by increased glucose oxidation may contribute to the amelioration of cardiac dysfunction in diabetes.

Regulation of NlE74A on vitellogenin may be mediated by angiotensin converting enzyme through a fecundity-related SNP in the brown planthopper, Nilaparvata lugens.

The major yolk protein precursors (YPP) gene, vitellogenin (Vg), usually considered as a reproductive indicator and molecular marker for evaluating insect fecundity, is controlled by insect hormone (mainly ecdysteroids and juvenile hormone), transcription factors and many other fecundity-related genes. To better understand the underlying molecular regulation mechanisms of the NlVg in the brown planthopper Nilaparvata lugens (N. lugens), the correlation between one early ecdysone response gene E74 and one important fecundity-related gene angiotensin converting enzyme (ACE) on the regulation of Vg gene expression, was investigated. We first showed that the mRNA expression level of NlACE were significantly higher in a high-fecundity population (HFP) than a low-fecundity population (LFP) at different development stages, and knockdown of NlACE expression by RNA interference (RNAi) results in a reduced level of NlVg expression and N. lugens fecundity. Subsequently, we analyzed the promoter of NlACE and found an E74A binding site, which was also differentially expressed in HFP and LFP. Then a gene putatively encoding E74A, namely NlE74A, predominant in the ovary and fat body was cloned and characterized. Furthermore, the developmental profile during female adult and the tissue-specific expression pattern of NlACE and NlE74A were similar to the expression pattern of NlVg gene, implying that both NlACE and NlE74A may be involved in regulating the expression of NlVg. Finally, after injecting the dsRNA of NlE74A, the NlACE expression levels were significantly reduced simultaneously at 24 h and 48 h post-injection, and the NlVg expression level was significant reduced at 24 h post-injection and the downswing was more significant at 48 h post-injection. These results imply that regulation of NlE74A on NlVg transcription might be mediated by NlACE through the E74 binding site at the NlACE promoter region in N. lugens.