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Rationale: Because of the lack of an antidote or effective treatment, patients with severe acute diquat and glyphosate poisoning always died within a few hours. Extracorporeal membrane pulmonary oxygenation (ECMO), as an artificial heart-lung supporting system, can be applied to support lung that is expected to recover from reversible pathological damage. However, to our knowledge, the application of ECMO for patients with diquat and glyphosate poisoning has not been reported. Patient concerns: A 40-year-old man ingested in 100 ml of diquat (20 g/100 ml) and 400 ml glyphosate (41 g/100 ml) was admitted to the intensive care unit (ICU), immediately complicated by the development of ventricular fibrillation, respiratory failure, renal failure, and multi-organ failure. Diagnosis: Diquat and glyphosate poisoning were diagnosed by stated ingestion history, and the diagnostic criteria for acute respiratory distress syndrome (ARDS) and multi-organ dysfunction syndrome were also met. Interventions: He was treated with veno-venous ECMO. Outcomes: He was successfully transferred out of the ICU on day 46 and discharged on day 67. The computed tomography scan showed no obvious pulmonary fibrosis 2 months after poisoning. Lessons: ECMO may be effective in the treatment of patients with severe ARDS caused by diquat and glyphosate poisoning when conventional management does not work.

Probiotic supplementation in early life may be effective in preventing atopic dermatitis (AD); however, results regarding efficacy have been controversial. The aim of our study was to investigate the effect of probiotic supplementation on the risk of AD. We systematically searched PubMed, EBSCO, Embase and Web of Science databases up to 8 March 2018 for potentially relevant studies regarding probiotic supplementation and AD. Included infants and children were those with probiotic exposure in utero and/or after birth who were not previously diagnosed with AD. We calculated the odds ratios (ORs) and 95% confidence intervals (CIs) and used the Jadad and Newcastle–Ottawa scales to assess methodologic quality. A total of 28 studies met the inclusion criteria. Compared with controls, probiotic treatment was associated with a reduced risk of AD (OR 0.69; 95% CI 0.58–0.82, P

The silk gland (SG) is characterized by the synthesis and secretion of silk protein in the economically important silkworm, Bombyx mori L. (Lepidoptera: Bombycidae). Nd and Nd-s are two fibroin-secretion-deficient silkworm mutants. MicroRNA (miRNA) plays an important role in many biological processes, such as cell proliferation, differentiation, and apoptosis. Using the Dazao silkworm as a control, we explored the miRNA expression profiles in the SGs of u02 (Nd) and u05 (Nd-s) to reveal the potential functions of miRNAs in silk protein expression and SG development. Here, we sequenced small RNA libraries made from the whole SGs of three strains. There are 260, 236, and 233 known miRNAs and 20, 18, and 18 potential new miRNAs identified from Dazao, u02, and u05, respectively. Fifty-three miRNAs are differentially expressed between Dazao and u02, 51 between Dazao and u05, and 16 between u02 and u05. Gene ontology/KEGG analyses show that most of the predicted target genes of differentially expressed miRNAs were assigned to functional categories involved in cell proliferation, organ development, and cellular compartment structures. The miRNA expression profile of naked silkworms will pave the way for the understanding of SG development and the regulation of silk protein expression.

Laryngeal squamous cell carcinoma (LSCC) is a common malignant tumor of the otolaryngeal region and accounts for 1-2% of all malignancies diagnosed worldwide. miR-340 down-regulation and EZH2 up-regulation have been frequently identified in multiple cancers, but the role of miR-340 and EZH2 in LSCC has not been explored. In this study, we investigated the regulative role of miR-340 in EZH2 expression and LSCC progression. The results showed that EZH2 was up-regulated and miR-340 was down-regulated in both Hep-2 cells and LSCC tissues. Molecularly, our results confirmed that miR-340 directly targeted EZH2 gene and inhibited EZH2 expression. MTT assay and BrdU assay showed that miR-340 transfection reduced the cell proliferation ability of Hep-2 cells. The transwell assay indicated that the invasion and migration ability of Hep-2 cells was dramatically inhibited by miR-340 transfection. In addition, miR-340 transfection induced cell apoptosis with concomitant enhancement of Bax, increase of Caspase-3 expression and activity, and reduction of Bcl-2 expression in Hep-2 cells. Both miR-340 transfection and EZH2 knockdown induced p27 expression and suppressed PI3K/Akt activation in Hep-2 cells. Strikingly, EZH2 knockdown reduced cell proliferation, and EZH2 overexpression significantly rescued the miR-340-mediated suppressive effect on cell proliferation. Moreover, miR-340 could obviously induce the inhibition of Hep-2 cell-derived tumor growth and EZH2/p27 expression ratio in vivo. Taken together, these data suggest that miR-340 impedes LSCC progression by targeting EZH2 with the possible mechanism to enhance the expression of anti-oncogene p27 and suppress PI3K/Akt activation, providing a novel target and a potential therapeutic pathway against LSCC.

Common cutworm (CCW; Spodoptera litura Fabricius) is a major leaf-eating insect that affects soybean crops in central and southern China. Understanding CCW-associated traits in soybean will provide insights for improved breeding strategies for resistance. The purpose of this study was to explore the genetic mechanisms underlying resistance to CCW. In this study, one natural population consisting of 219 soybean landraces from a wide geographic range was employed. Three traits, namely larval weight (LW), duration of larval stage (DLS) and pupal weight (PW) were evaluated as indicators for CCW resistance. Using 1536 single nucleotide polymorphisms (SNPs), a genome-wide association analysis was conducted to identify genomic regions associated with LW, DLS and PW in this natural population. Seven SNP-trait associations were identified using a mixed linear model (MLM). Four SNPs were co-associated with two traits. Three loci were significantly associated with CCW-resistance. Most of the detected loci were located within genomic regions previously identified as quantitative trait loci (QTLs) for these traits. The region on chromosome 6, which co-associated with LW and DLS, included several QTLs for reaction to disease or insect, such as Fusarium virguliforme, corn earworm and bean pyralid. On chromosome 7, two SNPs have been reported to associate with CCW, corn earworms, soybean aphid and bean pyralid moth infestations. Two putative genes (MYC2-like and JAM2-like) on chromosome 7 that may be associated with resistance to CCW were identified. These markers, located in previously reported QTLs, are consistent with previous findings and may represent important targets for the identification of CCW resistance-related genes. These results may provide a profound understanding of the genetic basis for resistance to CCW in soybean and the foundation for resistance to biotic stress through marker-assisted selection breeding strategies for CCW.

Plant architecture and yield components are critical for the determination of seed yield in soybean. In this study, we performed genetic association analysis to dissect the relationships between plant architecture and yield component traits. Two hundred and nineteen accessions were employed, and eight agronomic traits were evaluated in six environments. Our results revealed strong positive correlations of plant architecture traits with yield components and the significant association of 4 SNPs with plant architecture traits and of 7 SNPs with yield component traits in two or more environments. Eight SNPs were co-associated with two traits. Based on the phenotypic effects of the alleles of the detected SNPs, the best alleles were mined for twenty-three distinct SNPs. Fifteen typical carrier materials harbouring the best allele effects were also mined. Twenty parental combinations were proposed by pyramiding possible alleles per SNP in one individual (excluding possible epistatic effects). These proposed combinations with the best alleles and carrier materials will aid in the improvement of targeted traits and marker-assisted selection (MAS) efficiency in soybean breeding.

Soybean mosaic virus (SMV) disease is one of the most serious and broadly distributed soybean (Glycine max (L.) Merr.) diseases. Here, we combine the advantages of association and linkage analysis to identify and fine-map the soybean genes associated with resistance to SMV strain SC7. A set of 191 soybean accessions from different geographic origins and 184 recombinant inbred lines (RILs) derived from Kefeng No.1 (resistant) × Nannong 1138-2 (susceptible) were used in this study. The SC7 resistance genes were previously mapped to a 2.65 Mb region on chromosome 2 and a 380 kb region on chromosome 13. Among 19 single nucleotide polymorphisms (SNPs) detected via association analysis in the study, the SNP BARC-021625-04157 was located in the 2.65 Mb region, and the SNP BARC-041671-08065 was located near the 380 kb region; three genes harboring the SNPs were probably related to SC7 resistance. The resistance gene associated with BARC-021625-04157 was then fine-mapped to a region of approximately 158 kb on chromosome 2 using 184 RILs. Among the 15 genes within this region, one NBS-LRR type gene, one HSP40 gene and one serine carboxypeptidase-type gene might be candidate SC7 resistance genes. These results will be useful for map-based cloning and marker-assisted selection in soybean breeding programs.

Table S1. Hazard ratios associated with ACR and age subgroup for major adverse cardiovascular event (MACE). Table S2. Hazard ratios associated with ACR and age subgroup for cardiovascular death. Table S3. Hazard ratios associated with ACR and age subgroup for stroke. Table S4. Hazard ratios associated with ACR and age subgroup for myocardial infarction. (DOCX 38 kb)

Maternal genes present in mature oocytes play a crucial role in the early development of silkworm. Although maternal genes have been widely studied in many other species, there has been limited research in Bombyx mori. High-throughput next generation sequencing provides a practical method for gene discovery on a genome-wide level. Herein, a transcriptome study was used to identify maternal-related genes from silkworm eggs. Unfertilized eggs from five different stages of early development were used to detect the changing situation of gene expression. The expressed genes showed different patterns over time. Seventy-six maternal genes were annotated according to homology analysis with Drosophila melanogaster. More than half of the differentially expressed maternal genes fell into four expression patterns, while the expression patterns showed a downward trend over time. The functional annotation of these material genes was mainly related to transcription factor activity, growth factor activity, nucleic acid binding, RNA binding, ATP binding, and ion binding. Additionally, twenty-two gene clusters including maternal genes were identified from 18 scaffolds. Altogether, we plotted a profile for the maternal genes of Bombyx mori using a digital gene expression profiling method. This will provide the basis for maternal-specific signature research and improve the understanding of the early development of silkworm.

Objective: Head and neck squamous cell carcinoma (HNSCC) represents as a common malignancy with increasing incidence in the worldwide. The fact of its poor survival rate urgently requires developing efficient predictive biomarkers for clinical use. MicroRNAs (miRNAs) recently represent as a novel direction for early diagnosis and prognosis prediction in HNSCC therapy. In this study, we comprehensively investigated the function and putative target of miRNA-451 in vitro. Methods: The expression of miRNA-451 was detected in HNSCC tissues and cell lines by real-time PCR. Forced expression or inhibition of miRNA-451 was done by transient transfection of mimics or inhibitor of miRNA-451 into indicated cells, respectively. Cell proliferation was evaluated by cell counting and crystal staining. Afterwards, we perform western blot to verify the expression of the miRNA-451 predicted target, c-myc, after miRNA-451 was overexpressed. Results: We showed that miRNA-451 was downregulated in paired HNSCC tissues as well as in cell lines. And overexpression of miRNA-451 in cells with low endogenous expression of miRNA-451 accelerated proliferation. To the contrast, knockdown of miRNA-451 in cells with high levels of miRNA-451 significantly reduced cell growth rate. Furthermore, we used bioinformatics and cellular methods to predict and prove that c-myc was targeted by miRNA-451, since forced expression of miRNA-451 leaded to decreased c-myc protein expression in HNSCC cells. Conclusion: Our findings identify miRNA-451 as a potential biomarker and suggest a key role of miRNA-451-c-myc pathway in HNSCC cell transformation, which could represent a novel therapeutic strategy in HNSCC treatment.

A wafer‐scale high resolution patterning of bio‐microstructures is achieved using precise protein photolithography. In article number 1700191, the application of these micropatterned structures as cellular substrate for the successful spatial guidance of fetal neural stems cells using well‐defined silk fibroin light chain as the resist material is demonstrated by Tiger H. Tao and co‐workers.

Background & Aims Recent several studies suggested that nonsteroidal anti-inflammation drugs (NSAIDs) could prevent the pancreatitis after endoscopic retrograde cholangiopancreatography (ERCP). However, the routes of administration, the dosages of NSAIDs and the potential efficacy in reducing the severity of pancreatitis remain controversial. The aim of this meta-analysis was to evaluate the efficacy of NSAIDs for post-ERCP pancreatitis (PEP) prophylaxis. Methods We systematically searched PubMed, Embase, EBSCO, elsevier and web of science databases up to October 1, 2016 for relevant studies. Results A total of 24 studies were met the inclusion criteria. Compared to the controls, the risk of pancreatitis was much lower in the NSAIDs group (OR= 0.57, 95%CI: 0.48-0.67, P< 0.0001). However, NSAIDs were not effective in reducing the risk of moderate to severe pancreatitis compared with placebo (OR=0.75, 95%CI: 0.57-1.00). In the subanalyses, rectal administration was the only effective route (OR=0.51, 95%CI: 0.42-0.62), and the risk of PEP was reduced in both randomized controlled trials (RCTs) (OR=0.63, 95%CI: 0.52-0.76) and case-control articles (C-Cs) (OR=0.40, 95%CI: 0.28-0.58). Conclusions Prophylactic administration of NSAIDs reduced the incidence of PEP in both RCTs and C-Cs, especially when rectally administered, but was not effective in reducing the risk of moderate to severe pancreatitis. This article is protected by copyright. All rights reserved.

The traditional hybrid breeding techniques can no longer meet the increasing demands for silk production by the silkworm, Bombyx mori, and further improvement of the silk yield will depend on modern molecular breeding techniques. Here, we report improved silk yield in transgenic silkworms overexpressing the oncogene Yorkie

Precise patterning of biomaterials has widespread applications, including drug release, degradable implants, tissue engineering, and regenerative medicine. Patterning of protein‐based microstructures using UV‐photolithography has been demonstrated using protein as the resist material. The Achilles heel of existing protein‐based biophotoresists is the inevitable wide molecular weight distribution during the protein extraction/regeneration process, hindering their practical uses in the semiconductor industry where reliability and repeatability are paramount. A wafer‐scale high resolution patterning of bio‐microstructures using well‐defined silk fibroin light chain as the resist material is presented showing unprecedent performances. The lithographic and etching performance of silk fibroin light chain resists are evaluated systematically and the underlying mechanisms are thoroughly discussed. The micropatterned silk structures are tested as cellular substrates for the successful spatial guidance of fetal neural stems cells seeded on the patterned substrates. The enhanced patterning resolution, the improved etch resistance, and the inherent biocompatibility of such protein‐based photoresist provide new opportunities in fabricating large scale biocompatible functional microstructures.

Seed mineral elements are essential not only for seed germination and seedling morphological formation but also for human health. The genetic dissection of seed mineral accumulation is important for mineral biofortification in soybean. However, the molecular mechanisms controlling mineral element accumulation are genetically complex because a number of genetic loci are involved in the metabolic pathways of mineral accumulation in seeds. The objective of this study was to detect the genetic loci for mineral concentrations in soybean seeds, including calcium (Ca), magnesium (Mg), iron (Fe), zinc (Zn) and phosphorus (P). Quantitative trait loci (QTL) mapping for the corresponding traits was performed in 184 recombinant inbred lines (RILs) and 219 cultivated soybean accessions. The data for each year and the average across 2 years were used for identification and mapping of QTL controlling seed mineral concentrations. Linkage mapping in the RILs identified totally 35 significant QTL for the five seed mineral concentrations in three cases, some of which were co-localized. Association mapping in the 219 accessions detected 28 single-nucleotide polymorphisms associated with the seed mineral concentrations. Among these, BARC-018099-02516, which was associated with seed Zn concentration, was located close to qZn-11-2. There were 20 putative mineral-related genes in interesting regions of mineral QTL. Three QTL for seed weight were mapped in the RILs, two QTL for seed weight were co-localized with seed Ca, Zn and P concentration QTL which anchored to the same region on chromosome 11. These results will provide a profound understanding of the genetic basis for seed mineral accumulation in soybean seeds and the foundation for mineral biofortification through marker-assisted selection breeding.

ZnO nanorod array coatings were successfully synthesized on wood surfaces using a conventional hydrothermal (CHT) and a microwave-assisted hydrothermal (MWHT) method. The reaction time of the MWHT method at high temperature was only 1/12th that of the CHT method. The MWHT-ZnO nanorods exhibited better crystallinity than those produced by the CHT method. The ZnO nanorod had a wurtzite crystal structure oriented along the c-axis. The MWHT nanorods with their higher (002)/(101) ratio were more conducive and improved the orientation degree of the ZnO nanorods. Field emission scanning electron microscopy (FE-SEM) images showed that the MWHT-ZnO nanorods had smaller diameters and narrower size distributions than those produced by the CHT method. However, both methods formed well-aligned array coatings that covered the wood surfaces, and the two coatings had similar UV protecting effects and inhibited discoloration.

G protein-coupled receptor 40 (GPR40) is believed to be an attractive target to enhance insulin secretion in patients with type 2 diabetes. GPR40 has been found to couple to Gq protein, leading to the activation of phospholipase C and subsequent increases in the intracellular Ca(2+) level. However, the underlying mechanisms that regulate the internalization and desensitization of GPR40 remain to be elucidated. In the present study, a construct of GPR40 fused with enhanced green fluorescent protein (EGFP) at its C-terminus was constructed for direct imaging of the localization and internalization of GPR40 by confocal microscopy. In stably transfected HEK-293 cells, GPR40 receptors underwent rapid agonist-induced internalization and constitutive ligand-independent internalization. Our data demonstrated that the agonist-mediated internalization of GPR40 was significantly blocked by hypertonic sucrose treatment and by siRNA mediated depletion of the heavy chain of clathrin. In contrast, constitutive GPR40 internalization was not affected by hypertonic sucrose or by knock-down of clathrin expression, but it was affected by treatment with methyl-β-cyclodextrin (MβCD) and nystatin. Furthermore, our results using an arrestin-3-EGFP redistribution assay and siRNA-mediated knock-down of arrestin-3 and GRK2 expression revealed that arrestin-3 and GRK2 play an essential role in the regulation of agonist-mediated GPR40 internalization, but are not involved in the regulation of constitutive GPR40 internalization. Additionally, our observation showed that upon activation by agonist, the internalized GPR40 receptors were rapidly recycled back to the plasma membrane via Rab4/Rab5 positive endosomes, whereas the constitutively internalized GPR40 receptors were recycled back to the cell surface through Rab5 positive endosomes. Because FFA receptors exhibit a high level of homology, our observations could be applicable to other members of this family.

Tachykinins constitute one of the largest peptide families in the animal kingdom and exert their diverse actions via G protein-coupled receptors (GPCRs). In this study, the Bombyx tachykinin-related peptides (TKRPs) were identified as specific endogenous ligands for the Bombyx neuropeptide GPCR A24 (BNGR-A24) and thus designated BNGR-A24 as BmTKRPR. Using both mammalian cell line HEK293 and insect cell line Sf21, further characterization demonstrated that BmTKRPR was activated, thus resulting in intracellular accumulation of cAMP, Ca(2+) mobilization, and ERK1/2 phosphorylation in a Gs and Gq inhibitor-sensitive manner. Moreover, quantitative reverse transcriptase polymerase chain reaction analysis and dsRNA-mediated knockdown experiments suggested a possible role for BmTKRPR in the regulation of feeding and growth. Our findings enhance the understanding of the Bombyx TKRP system in the regulation of fundamental physiological processes.

To evaluate the genetic diversity (GD) of wild and cultivated soybeans and determine the genetic relationships between them, in this study, 127 wild soybean accessions and 219 cultivated soybean accessions were genotyped using 74 simple sequence repeat (SSR) markers. The results of the study revealed that the GD of the wild soybeans exceeded that of the cultivated soybeans. In all, 924 alleles were detected in the 346 soybean accessions using 74 SSRs, with an average of 12.49 alleles per locus. In the 219 cultivated soybean accessions, 687 alleles were detected, with an average of 9.28 alleles per locus; in the 127 wild soybean accessions, 835 alleles were detected, with an average of 11.28 alleles per locus. We identified 237 wild-soybean-specific alleles and 89 cultivated-soybean-specific alleles in the 346 soybean accessions, and these alleles accounted for 35.28% of all the alleles in the sample. Principal coordinates analysis and phylogenetic analysis based on Nei's genetic distance indicated that all the accessions could be classified into two major clusters, corresponding to wild and cultivated soybeans. These results will increase our understanding of the genetic differences and relationships between wild and cultivated soybeans and provide information to develop future breeding strategies to improve soybean yield.

DNJ, an inhibitor of α-glucosidase, is used to suppress the elevation of postprandial hyperglycemia. In this study, we focus on screening an appropriate microorganism for performing fermentation to improve DNJ content in mulberry leaf. Results showed that Ganoderma lucidum was selected from 8 species and shown to be the most effective in improvement of DNJ production from mulberry leaves through fermentation. Based on single factor and three factor influence level tests by following the Plackett-Burman design, the optimum extraction yield was analyzed by response surface methodology (RSM). The extracted DNJ was determined by reverse-phase high performance liquid chromatograph equipped with fluorescence detector (HPLC-FD). The results of RSM showed that the optimal condition for mulberry fermentation was defined as pH 6.97, potassium nitrate content 0.81% and inoculums volume 2 mL. The extraction efficiency reached to 0.548% in maximum which is 2.74 fold of those in mulberry leaf.

Bombyx mori cathepsin B (BmCatB) is involved in the programmed cell death of the fat body during B. mori metamorphosis. For a better understanding of the functional regulatory mechanism, the promoter region of BmCatB in the transcriptional regulation has been identified and analyzed in the present study. BmCatB promoter region performed by the 5' truncation or mutagenesis of EcREs was inserted in the pFA3Luc-A3RL double fluorescence expression vector to activate the fireflies luciferase (FLuc) gene. The results indicated that the dual-luciferase activity of BmCatB gene in the silkworm larval fat body is regulated by the length of promoter. Site-directed mutagenesis of EcRE experiment has shown that the EcREs are up-regulated significantly in the regulation of the BmCatB promoter. A 142bp region (-1165 to -1023) and EcREs are the mainly fat-body tissue-specificity related region and could function as a core promoter element.

In materials science and engineering, designed and well-ordered self-assembled hydrogel with excellent biocompatibility and nontoxic properties is an inviting issue. In this study, a novel self-assembled sericin hydrogel was prepared by a simple-two-step process containing milling and high temperature and high pressure. It is worthwhile pointing out that the sericin with high extraction rate (95%) for fabricating sericin hydrogel was isolated from fibroin-deficient mutant silk cocoons. The hydrogel was formed under a mild condition without introducing any other toxic or harmful substances and its gelling time could be controlled by sericin concentration, temperature or pH values. Moreover, as long as the sericin solution is kept in a bioclean circumstance, the sterile sericin hydrogel can be achieved and no need to sterilize. Furthermore, this hydrogel possesses high porosity, degradation. In addition, the hydrogel system is suitable as a carrier of cells or bioactive molecules contribute from its excellent cell-adhesive capability, effectively promoting cell attachment, proliferation, and a sustained manner for drug release. In summary, our study indicates that the self-assembly sericin hydrogel could be promising candidates in soft tissue engineering applications.

Aimed at reducing the switching loss and common-mode voltage amplitude of high-power medium-voltage three-level inverter under low modulation index conditions, an improved synchronous space vector PWM strategy is proposed in this paper. The switching times in each fundamental period are reduced by the re-division of small regions and the full use of the redundant switching state. The sum of switching algebra is introduced as an evaluation index and the switching state with the minimum value of the sum of switching algebra are adopted. Then, the common mode voltage amplitude is reduced. The theoretical analysis and experimental results show that the improved modulation strategy proposed in this paper can effectively reduce the switching loss and common-mode voltage amplitude of the inverter under the condition of the low modulation index. Moreover, the neutral-point voltage ripple is also reduced simultaneously.

A high optically transparent pure sericin hydrogel without any harmful or toxic ingredients is attractive for cell culture system and drug delivery. Here we report a novel 3D pure sericin hydrogel fabricated using ultrasonication method. Gelation time of the hydrogel could be controlled from minutes to days by regulated sonication parameters, including power output and time, as well as sericin concentration. This hydrogel possesses high optically transparence, injectability, eximious elasticity, pH-dependent degradation as well as high porosity. In addition, the hydrogel system is compatible for hosting cell and bioactive molecules owing to its excellent cell-adhesive capability, effectively promoting cell attachment, proliferation and long-term survival, along with delivering bioactive reagent in a sustained manner. Together, our study demonstrates that the sericin hydrogel may serve as a multifunctional platform for cell culture and drug delivery.

The "Ming" lethal egg mutant (l-em) is a vitelline membrane mutant in silkworm, Bombyx mori. The eggs laid by the l-em mutant lose water, ultimately causing death within an hour. Previous studies have shown that the deletion of BmEP80 is responsible for the l-em mutation in silkworm, B. mori. In the current study, digital gene expression (DGE) was performed to investigate the difference of gene expression in ovaries between wild type and l-em mutant on the sixth day of the pupal stage to obtain a global view of gene expression profiles using the ovaries of three l-em mutants and three wild types. The results showed a total of 3,463,495 and 3,607,936 clean tags in the wild type and the l-em mutant libraries, respectively. Compared with those of wild type, 239 differentially expressed genes were detected in the l-em mutant, wherein 181 genes are up-regulated and 58 genes are down-regulated in the mutant strain. The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis results showed that no pathway was significantly enriched and three pathways are tightly related to protein synthesis among the five leading pathways. Moreover, the expression profiles of eight important differentially expressed genes related to oogenesis changed. These results provide a comprehensive gene expression analysis of oogenesis and vitellogenesis in B. mori which facilitates understanding of both the specific molecular mechanism of the 1-em mutant and Lepidopteran oogenesis in general.

The egg stage is an important stage in the silkworm (Bombyx mori) life cycle. Normal silkworm eggs are usually short, elliptical, and laterally flattened, with a sometimes hollowed surface on the lateral side. However, the eggs laid by homozygous recessive "Ming" lethal egg mutants (l-e(m)) lose water and become concaved around 1h, ultimately exhibiting a triangular shape on the egg surfaces. We performed positional cloning, and narrowed down the region containing the gene responsible for the l-e(m) mutant to 360 kb on chromosome 10 using 2287 F(2) individuals. Using expression analysis and RNA interference, the best l-e(m) candidate gene was shown to be BmEP80. The results of the inverse polymerase chain reaction showed that an ~1.9 kb region from the 3' untranslated region of BmVMP23 to the forepart of BmEP80 was replaced by a100 kb DNA fragment in the l-e(m) mutant. Several eggs laid by the normal moths injected with BmEP80 small interfering RNAs were evidently depressed and exhibited a triangular shape on the surface. The phenotype exhibited was consistent with the eggs laid by the l-e(m) mutant. Moreover, two-dimensional gel electrophoresis showed that the BmEP80 protein was expressed in the ovary from the 9th day of the pupa stage to eclosion in the wild-type silkworm, but was absent in the l-e(m) mutant. These results indicate that BmEP80 is responsible for the l-e(m) mutation.

In this study, we report a novel cellulase [β-1,4-endoglucanase (EGase), EC 3.2.1.4] cDNA (Bh-EGase II) belonging to the glycoside hydrolase family (GHF) 45 from the beetle Batocera horsfieldi. The Bh-EGase II gene spans 720 bp and consists of a single exon coding for 239 amino acid residues. Bh-EGase II showed 93.72% protein sequence identity to Ag-EGase II from the beetle Apriona germari. The GHF 45 catalytic site is conserved in Bh-EGase II. Bh-EGase II has three putative N-glycosylation sites at 56–58 (N–K–S), 99–101 (N–S–T), and 237–239 (N–Y–S), respectively. The cDNA encoding Bh-EGase II was expressed in baculovirus-infected insect BmN cells and Bombyx mori larvae. Recombinant Bh-EGase II from BmN cells and larval hemolymph had an enzymatic activity of approximately 928 U/mg. The enzymatic catalysis of recombinant Bh-EGase II showed the highest activity at 50 °C and pH 6.0.

Eclampsia is a hypertensive disorder of pregnancy that is defined by the new onset of grand mal seizures on the basis of preeclampsia and a leading cause of maternal and fetal mortality worldwide. Presently, magnesium sulfate (MgSO4) is the most effective treatment, but the mechanism by which MgSO4 prevents eclampsia has yet to be fully elucidated. We previously showed that systemic inflammation decreases the seizure threshold in a rat eclampsia-like model, and MgSO4 treatment can decrease systemic inflammation. Here, we hypothesized that MgSO4 plays a neuroprotective role in eclampsia by reducing neuroinflammation and brain edema. Pregnant Sprague-Dawley rats were given an intraperitoneal injection of pentylenetetrazol following a tail vein injection of lipopolysaccharide to establish the eclampsia-like seizure model. Seizure activity was assessed by behavioral testing. Neuronal loss in the hippocampal CA1 region (CA1) was detected by Nissl staining. Cerebrospinal fluid levels of S100-B and ferritin, indicators of neuroinflammation, were detected by enzyme-linked immunosorbent assay, and ionized calcium binder adapter molecule 1 (Iba-1, a marker for microglia) and glial fibrillary acid protein (GFAP, a marker for astrocytes) expression in the CA1 area was determined by immunofluorescence staining. Brain edema was measured. Our results revealed that MgSO4 effectively attenuated seizure severity and CA1 neuronal loss. In addition, MgSO4 significantly reduced cerebrospinal fluid levels of S100-B and ferritin, Iba-1 and GFAP activation in the CA1 area, and brain edema. Our results indicate that MgSO4 plays a neuroprotective role against eclampsia-like seizure by reducing neuroinflammation and brain edema.

MicroRNAs (miRNAs) are small endogenous RNAs molecules, approximately 21–23 nucleotides in length, which regulate gene expression by base-pairing with 3′ untranslated regions (UTRs) of target mRNAs. However, the functions of only a few miRNAs in organisms are known. Recently, the expression vector of artificial miRNA has become a promising tool for gene function studies. Here, a method for easy and rapid construction of eukaryotic miRNA expression vector was described. The cytoplasmic actin 3 (A3) promoter and flanked sequences of miRNA-9a (miR-9a) precursor were amplified from genomic DNA of the silkworm (Bombyx mori) and was inserted into pCDNA3.0 vector to construct a recombinant plasmid. The enhanced green fluorescent protein (EGFP) gene was used as reporter gene. The Bombyx mori N (BmN) cells were transfected with recombinant miR-9a expression plasmid and were harvested 48 h post transfection. Total RNAs of BmN cells transfected with recombinant vectors were extracted and the expression of miR-9a was evaluated by reverse transcriptase polymerase chain reaction (RT-PCR) and Northern blot. Tests showed that the recombinant miR-9a vector was successfully constructed and the expression of miR-9a with EGFP was detected.

Filamentation temperature-sensitive H (FtsH) is an ATP-dependent zinc metalloprotease involved in diverse biological functions. There are 12 FtsH proteins in Arabidopsis, among which AtFtsH2 plays an important role in regulating the turnover of photosystem II (PSII) reaction center D1 protein and the development of the photosynthetic apparatus. Here, we have identified 11 FtsH genes in the soybean genome by a bioinformatics approach. These soybean FtsH genes corresponded to seven Arabidopsis FtsH genes, suggesting that the main characteristics of soybean FtsH genes were formed before the evolutionary split of soybean and Arabidopsis. Phylogenetic analyses allowed us to clone a soybean AtFtsH2-like gene designated as GmFtsH9. The predicted protein of GmFtsH9 consists of 690 amino acids and contains three typical FtsH proteins conserved domains. The expression level of GmFtsH9 was determined in a soybean recombinant inbred line population under a pot experiment conducted for measuring chlorophyll a fluorescence transient parameters, photosynthetic CO(2) fixation rate (P (N)), and seed yield. Expression quantitative trait loci (eQTL) mapping revealed two trans-acting eQTLs for GmFtsH9. The significant correlation of gene expression level with chlorophyll a fluorescence transient parameters and the presence of overlapping eQTL (QTL) between gene expression level and chlorophyll a fluorescence transient parameters indicated that GmFtsH9 could be involved in regulating PSII function. These results further lead to the understanding of the mechanism underlying FtsH gene expression, and contribute to the development of marker-assisted selection breeding programs for modulating soybean FtsH gene expression.

G protein-coupled receptors (GPCRs) are the largest and most versatile family of transmembrane receptors in the cell, occupying the highest hierarchical positions in the regulation of many physiological processes. Although they have been extensively studied in a number of model insects, there have been few investigations of GPCRs in large Lepidopterans, such as Bombyx mori, an organism that provides a means to perform detailed tissue expression analyses, which may help to characterize GPCRs and their ligands. In addition, B. mori, also known as the silkworm, is an insect of substantial economic importance, due to its use in silk production and traditional medicines. In this work, we computationally identified 90 putative GPCRs in B. mori, 33 of which represent novel proteins. These GPCRs were annotated and compared with their homologs in Drosophila melanogaster and Anopheles gambiae. Phylogenetics analyses of the GPCRs from these three insects showed that GPCRs may easily duplicate or disappear during insect evolution, especially in the neuropeptide and protein hormone receptor subfamily. Interestingly, we observed a decrease in the quantity and diversity of the stress-tolerance gene, Methuselah, in B. mori, which may be related to its long history of domestication. Moreover, the presence of many Bombyx-specific GPCRs suggests that neither Drosophila nor Anopheles is good representatives for the GPCRs in the Class Insecta.

Kinetic studies were performed for the chelate ring closing and opening process of cyclopentadienyl cobalt(III) complexes having a pendant N-functional group with an amine, piperidine or pyridine moiety in the side chain. The metal–nitrogen bond energy was measured. The rate of chelation by such pendant N-functionalized side chains in diiodomonocarbonyl cobalt(III) reaction intermediates is determined by the electronic density on the donor atom and the strength of the forming chelated bond. The steric factor around the donor atom plays a secondary role. On the basis of the enthalpies and entropies obtained from the kinetic studies, the process of chelate ring closing in diiodomonocarbonyl cobalt(III) reaction intermediates is via an associative pathway involving loss of CO, while the chelate ring opening process in the resulted chelators is via a metal–nitrogen bond cleavage, solvation then metal–phosphorus bond formation pathway during substitution of PPh 3 . The chelator with the most rigid arm of picolyl shows a smallest steric hindrance for incoming PPh 3 compared to the other two analogues.

To investigate effects of atmospheric pollutants on fog nature, a comprehensive in situ observation project was implemented in the northern suburb of Nanjing, in December of 2006. For December 24–27 there occurred a heavy fog lasting 4 d in succession. This event is of rare characteristics, namely long persistence, high concentration, tall fog top, acid fog water and explosive growth. Detailed analysis along with the causes of the fog was presented. The evidence suggests that the fog was generated by nighttime radiative cooling, maintained and developed under effects of warm, wet advection. As a result, it is an advection-radiation fog event.

In the silkworm, metamorphosis and moulting are regulated by ecdysone hormone and juvenile hormone. The subject in the present study is a silkworm mutant that does not moult in the 2nd instar (nm2). Genetic analysis indicated that the nm2 mutation is controlled by a recessive gene and is homozygous lethal. Based on positional cloning, nm2 was located in a region approximately 275 kb on the 5th linkage group by eleven SSR polymorphism markers. In this specific range, according to the transcriptional expression of thirteen genes and cloning, the relative expression level of the BmCPG10 gene that encodes a cuticle protein was lower than the expression level of the wild-type gene. Moreover, this gene's structure differs from that of the wild-type gene: there is a deletion of 217 bp in its open reading frame, which resulted in a change in the protein it encoded. The BmCPG10 mRNA was detectable throughout silkworm development from the egg to the moth. This mRNA was low in the pre-moulting and moulting stages of each instar but was high in the gluttonous stage and in newly exuviated larvae. The BmCPG10 mRNA showed high expression levels in the epidermis, head and trachea, while the expression levels were low in the midgut, Malpighian tubule, prothoracic gland, haemolymph and ventral nerve cord. The ecdysone titre was determined by ELISA, and the results demonstrated that the ecdysone titre of nm2 larvae was lower than that of the wild-type larvae. The nm2 mutant could be rescued by feeding 20-hydroxyecdysone, cholesterol and 7-dehydrocholesterol (7dC), but the rescued nm2 only developed to the 4th instar and subsequently died. The moulting time of silkworms could be delayed by BmCPG10 RNAi. Thus, we speculated that the mutation of BmCPG10 was responsible for the silkworm mutant that did not moult in the 2nd instar.

As revealed in a previous microarray study to identify genes regulated by 20-hydroxyecdysone (20E) and juvenile hormone (JH) in the silkworm, Bombyx mori, E93 expression in the fat body was markedly low prior to the wandering stage but abundant during larval-pupal metamorphosis. Induced by 20E and suppressed by JH, E93 expression follows this developmental profile in multiple silkworm alleles. The reduction of E93 expression by RNAi disrupted 20E signaling and the 20E-induced autophagy, caspase activity, and cell dissociation in the fat body. Reducing E93 expression also decreased the expression of the 20E-induced pupal-specific cuticle protein genes and prevented growth and differentiation of the wing discs. Importantly, the two HTH domains in E93 are critical for inducing the expression of a subset of 20E response genes, including EcR, USP, E74, Br-C, and Atg1. By contrast, the LLQHLL and PLDLSAK motifs in E93 inhibit its transcriptional activity. E93 binds to the EcR-USP complex via a physical association with USP through its LLQHLL motif; and this association is enhanced by 20E-induced EcR-USP interaction, which attenuates the transcriptional activity of E93. E93 acts through the two HTH domains to bind to GAGA-containing motifs present in the Atg1 promoter region for inducing gene expression. In conclusion, E93 transcriptionally modulates 20E signaling to promote Bombyx larval-pupal metamorphosis.

Tolerance to low-phosphorus soil is a desirable trait in soybean cultivars. Previous quantitative trait locus (QTL) studies for phosphorus-deficiency tolerance were mainly derived from bi-parental segregating populations and few reports from natural population. The objective of this study was to detect QTLs that regulate phosphorus-deficiency tolerance in soybean using association mapping approach. Phosphorus-deficiency tolerance was evaluated according to five traits (plant shoot height, shoot dry weight, phosphorus concentration, phosphorus acquisition efficiency and use efficiency) comprising a conditional phenotype at the seedling stage. Association mapping of the conditional phenotype detected 19 SNPs including 13 SNPs that were significantly associated with the five traits across two years. A novel cluster of SNPs, including three SNPs that consistently showed significant effects over two years, that associated with more than one trait was detected on chromosome 3. All favorable alleles, which were determined based on the mean of conditional phenotypic values of each trait over the two years, could be pyramided into one cultivar through parental cross combination. The best three cross combinations were predicted with the aim of simultaneously improving phosphorus acquisition efficiency and use efficiency. These results will provide a thorough understanding of the genetic basis of phosphorus deficiency tolerance in soybean.

To investigate the effect of patient-controlled epidural analgesia (PCEA) on uterine electromyography (EMG) activity in term pregnant women during labor.Nulliparous pregnant women in spontaneous term labor (N = 30) were enrolled (PCEA group, n = 20 and control group, n = 10). Five time periods (30 minutes each) were defined for noninvasive abdominal recordings and analysis of uterine EMG activity, that is, period I: before PCEA treatment with 2-cm cervical dilation; periods II to IV: each period successively at 30, 60, and 120 minutes after PCEA; and period V: second stage of labor with cervix at 10 cm dilation. Control patients without PCEA were monitored during the same times. The number of bursts/30 min, power density spectrum peak frequency, mean amplitude, and duration of uterine EMG bursts were measured to assess uterine EMG activity. Maternal, fetal, and labor characteristics were also recorded. Data were analyzed by analysis of variance followed by other tests.Electromyography parameters are significantly lower (P.001) after PCEA (periods II to IV) compared to controls but similar between groups by period V (P.05). Also, patients with PCEA have a slower rate of cervical dilation (P.003, period IV only) and longer labor in both stage 1 and stage 2 (P.05). All patients have similar (P.05) positive labor outcomes.Patient-controlled epidural analgesia initially suppresses uterine EMG and slows cervical dilation thereby prolonging labor. However, the EMG activity recovers with labor progress with no effects on delivery outcomes.

Wood-feeding insects depend heavily on the secretion of a combination of cellulases, mainly endoglucanases and other glucanases such as exoglucanases and xylanases, to achieve efficient digestion of the cellulose of cellulosic materials. In this paper, we report a novel cellulose Bh-EGaseI belonging to the glycoside hydrolase family 45(gh45-1) obtained from the beetle Batocera horsfieldi. The Bh-EGaseI gene spans 714 bp and consists of three exons coding 237 amino acid residues. The cDNA encoding Bh-EGaseI was expressed as 25 KDa in baculovirus-infected Bombyx mori larvae. The expression products of Bh-EGaseI from larval hemolymph showed a specific enzymatic activity of approximately 1030.87 IU per mg. The enzyme was active over a wide range of pH and temperatures; optimal activity was observed at 40 °C and pH 4.0. The effects of ions on Bh-EGaseI activity were also studied, and results indicated that activity decreased to different extents upon addition of ions. Investigations on Bh-EGaseI facilitate their potential application in the production of bioenergy and biomaterials from cellulosic biomass in the future.

Cranial fixation should be safe, reliable, ideally degradable, and produce no hazardous residues and no artifacts on neuroimaging. Protein-based fixation devices offer an exciting opportunity for this application. Here, the preclinical development and in vivo efficacy verification of a silk cranial fixation system in functional models are reported by addressing key challenges toward clinical use. A comprehensive study on this fixation system in rodent and canine animal models for up to 12 months is carried out. The silk fixation system shows a superb performance on the long-term stability of the internal structural support for cranial flap fixation and bone reconnection and has good magnetic resonance imaging compatibility, and tolerability to high dose radiotherapy, underscoring the favorable clinical application of this system for neurosurgery compared to the current gold standard.

Precise patterning of biomaterials has widespread applications, including drug release, degradable implants, tissue engineering, and regenerative medicine. Patterning of protein-based microstructures using UV-photolithography has been demonstrated using protein as the resist material. The Achilles heel of existing protein-based biophotoresists is the inevitable wide molecular weight distribution during the protein extraction/regeneration process, hindering their practical uses in the semiconductor industry where reliability and repeatability are paramount. A wafer-scale high resolution patterning of bio-microstructures using well-defined silk fibroin light chain as the resist material is presented showing unprecedent performances. The lithographic and etching performance of silk fibroin light chain resists are evaluated systematically and the underlying mechanisms are thoroughly discussed. The micropatterned silk structures are tested as cellular substrates for the successful spatial guidance of fetal neural stems cells seeded on the patterned substrates. The enhanced patterning resolution, the improved etch resistance, and the inherent biocompatibility of such protein-based photoresist provide new opportunities in fabricating large scale biocompatible functional microstructures.

To analyze the effect of delayed hemorrhage after endoscopic sphincterotomy (EST) and compare the efficacy in improving complication between medicine treatment alone and medicine combined with endoscopic treatment. 1741 patients with EST admitted in Yijishan hospital of Wannan medical college from September 2009 to May 2014 were enrolled in this study. 32 cases suffered from delayed hemorrhage. The patients with delayed hemorrhage were evaluated through incision length of duodenal papilla, clinical manifestation, stool occult blood test and the difference of hemoglobin concentration between pre and post operation. 32 patients were divided into mild bleeding group, mild serious group and serious group through the speed and amount of bleeding. All cases in mild group accepted medicine treatment. Mild serious group were divided into medicine therapy group and medicine combined with endoscopic therapy group randomly. Serious group accepted vascular intervention therapy even traditional operation. The different treatments for delayed hemorrhage were judged by efficiency. The dates were analyzed by t-test or chi-square test. Nobody endured delayed hemorrhage who accepted small incision. Delayed hemorrhage was found in 7 patients out of 627 cases who accepted medium-large incision, 25 patients of 920 cases who accepted large incision. The patients who accepted lager EST were more dangerous than small EST (χ(2)=4.718, P=0.030) concerning delayed hemorrhage. 32 cases in 1741 patients suffered from delayed hemorrhage. 14 patients only have passed black stool after EST. Among 14 cases, 13 patients stop bleeding after medical therapy, and 1 case received endoscopic hemostasis. 15 cases with hematemesis or melena after EST, 7 patients who received combination therapy stop bleeding. 3 patients from 8 cases stop bleeding after single chemical treatment, 5 cases had to receive endoscopic hemostasis after ineffectual medical therapy. There are significant difference for concerning effect between combination therapy group and medical therapy group (P=0.026). 3 patients repeatedly vomited blood and develop to peripheral circulatory failure. Those patients all received vascular intervention therapy, 2 patients stop bleeding, 1 patient failed in vascular intervention therapy and given up emergency rescue and died. Large EST has more risks than small EST in concerning delayed hemorrhage. Delayed bleeding after EST should be treated by different levels. Adapted therapy should be recommend for patients with different levels bleeding.

Sericin, a major component of silk, has a long history of being discarded as a waste during silk processing. The value of sericin for tissue engineering is underestimated and its potential application in regenerative medicine has just begun to be explored. Here we report the successful fabrication and characterization of a covalently-crosslinked 3D pure sericin hydrogel for delivery of cells and drugs. This hydrogel is injectable, permitting its implantation through minimally invasive approaches. Notably, this hydrogel is found to exhibit photoluminescence, enabling bioimaging and in vivo tracking. Moreover, this hydrogel system possesses excellent cell-adhesive capability, effectively promoting cell attachment, proliferation and long-term survival of various types of cells. Further, the sericin hydrogel releases bioactive reagents in a sustained manner. Additionally, this hydrogel demonstrates good elasticity, high porosity, and pH-dependent degradation dynamics, which are advantageous for this sericin hydrogel to serve as a delivery vehicle for cells and therapeutic drugs. With all these unique features, it is expected that this sericin hydrogel will have wide utility in the areas of tissue engineering and regenerative medicine.

Neutral-point (NP) voltage oscillation amplitude, THD of output voltage and the switching frequency of the inverter are the three key performance indices in three-level neutral-point-clamped (NPC) inverter. Every element connects with each other and restricts each other, which indicates that to keep the three key performance indices balanced is the research focus of the modulation strategy for three-level NPC inverter. Hybrid space vector PWM strategy, which is proposed in this paper, combines NV and non-NV, synthesizes the advantages of the two methods, reduces NP low-frequency oscillation and also decreases switching frequency and output voltage THD. Taking NTV (the most typical method of NV) and NTV (the most typical method of non-NV) as examples, A new hybrid SVPWM method, based on the NP current maximum minimum (MM) method is proposed in this paper. This new hybrid space vector PWM method applies NTV to uncontrolled balancing area and minimizes the switching frequency and THD of output voltage within the range of the allowable NP oscillation amplitude. Meanwhile, the impact on the output voltage produced by NP oscillation is researched in this paper. The experimental results show the validity of the proposed strategy.

BACKGROUND: Despite the tight correlation between T-type Ca2+ channels and a great variety of tumors, the roles of alpha1G subunit of T-type Ca2+ channels in laryngeal squamous cell carcinoma (LSCC) have not yet been investigated. METHODS: In the present study, we examined the expression of alpha1G subunit of T-type Ca2+ channel in human LSCC tissues and cell lines. One human laryngeal squamous cell carcinoma cell line, Hep-2, was also examined for T-type channels using voltage-clamp recordings. Cell proliferation assays were performed in the presence or absence of T-type channel blocker mibefradil and alpha1G subunit sepcific siRNA. The cell cycle was determined by flow cytometry. RESULTS: Our results indicated that the a1G subunit of T-type Ca2+ channel is highly expressed in human laryngeal squamous cell carcinoma tissues and cell lines. alpha1G siRNA significantly down-regulated the protein expression of the alpha1G subunit. Both alpha1G siRNA and mibefradil inhibited Hep-2 cell proliferation and arrested cell cycle progression. CONCLUSIONS: Together, these findings suggest a functional role of T-type channels in certain laryngeal carcinomas, and that inhibition of T-type channels reduces cell proliferation via cell cycle arrest, suggesting that the alpha1G subunit of T-type Ca2+ channel may be used as a therapeutic target for treating LSCC.

White egg 2 is one of white egg mutants in silkworm, whose molecular mechanism remains unknown so far. In order to obtain an overall view on gene expression profiles at early embryo development stages, the white egg 2 near-isogenic line was constructed and the whole-genome of silkworm microarray system containing 21375 predicted genes from the silkworm whole genome sequence was employed to investigate gene expression profiles at 0, 24 and 48 h post oviposition between white egg 2 mutant and normal black egg strain. At 24 h post oviposition, 49 genes exhibited at least 2.0 fold differences at expression level, including 24 up-regulated genes and 25 down-regulated genes while at 48 h post oviposition, 52 genes, including 23 up-regulated genes and 29 down-regulated genes were expressed differentially over 2.0 change fold. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that nine differentially expression genes were involved in nine significant (p

To evaluate the pilots' safety behaviors for airline flight, an evaluation method Based on fuzzy neural network is investigated. Firstly, an evaluation index system of airline pilots' safety behaviors under the background of Chinese aviation safety culture is established. Secondly, for the first time, on the basis of combing fuzzy theory and neural network, the improved fuzzy neural network technology is applied to evaluate airline pilots' safety behaviors. Finally, a case study is put in practice, and simulation results through testing model show that the proposed method can evaluate airline pilots' safety behaviors effectively, also it will be very helpful for airline companies' choosing and training airline pilots in the future.

Members of the mammalian neuropeptide Y (NPY) family serve as neurotransmitters and contribute to a diversity of physiological functions. Although neuropeptide F (NPF), the NPY-like orthologs from insects, have been identified, the NPF receptors and their signaling and physiological functions remain largely unknown. In this study, we established the stable and transient functional expression of a Bombyx orphan G protein-coupled receptor, BNGR-A4, in both mammalian HEK293 and insect SF21 cells. We identified Bombyx mori NPFs as specific endogenous ligands for the Bombyx Neuropeptide GPCR A4 (BNGR-A4) and, accordingly, named the receptor BomNPFR. Our results demonstrated that BomNPFR was activated by synthetic BomNPF1a and BomNPF1b at a high efficacy and by BomNPF2 at a low efficacy. This activation led to a decrease of forskolin or adipokinetic hormone peptide-stimulated adenylyl cyclase activity, an increase of intracellular Ca(2+), the activation of ERK1/2 signaling and receptor internalization. Moreover, a Rhodamine-labeled BomNPF1a peptide was found to bind specifically to BomNPFR. The results derived from quantitative RT-PCR analysis and dsRNA-mediated knockdown experiments demonstrated the possible role of BomNPFR in the regulation of food intake and growth. Our results provide the first in-depth information on BomNPFR-mediated signaling for the further elucidation of the BomNPF/BomNPFR system in the regulation of fundamental physiological processes.