miR-317-3p and miR-283-5p Play a Crucial Role in Regulating the Resistance to Indoxacarb in Spodoptera frugiperda by Targeting GSTs4.

Spodoptera frugiperda is primarily controlled through chemical insecticides. Our RNA-seq data highlight the overexpression of GSTs4 in indoxacarb-resistant S. frugiperda. However, the exact role of GSTs4 in indoxacarb resistance and its regulatory mechanisms remains elusive. Therefore, we investigated the functional role of GSTs4 in S. frugiperda and explored the underlying post-transcriptional regulatory mechanisms. GSTs4 was highly overexpressed (27.6-fold) in the indoxacarb-resistant strain, and GSTs4 silencing significantly increases the susceptibility of S. frugiperda to indoxacarb, increasing mortality by 27.3%. miR-317-3p and miR-283-5p can bind to the 3'UTR of GSTs4, and the targeting relationship was confirmed by dual-luciferase reporter assays. Injecting miR-317-3p and miR-283-5p agomirs reduces GSTs4 levels by 64.8 and 42.3%, respectively, resulting in an increased susceptibility of S. frugiperda to indoxacarb. Conversely, the administration of miR-317-3p and miR-283-5pantagomirs increases GSTs4 expression and reduces larval susceptibility to indoxacarb. These findings demonstrate that miR-317-3p and miR-283-5p contribute to indoxacarb resistance in S. frugiperda by regulating the overexpression of GSTs4.

Sublethal effects of acetamiprid and afidopyropen on Harmonia axyridis: insights from transcriptomics analysis.

Evaluating the sublethal effects of insecticide is crucial for protecting and utilizing natural enemies. In this study, we determined the sublethal effects of acetamiprid and afidopyropen on Harmonia axyridis (Pallas) and explored the potential molecular mechanisms underlying these effects through transcriptomics analysis. The results showed that sublethal concentrations of acetamiprid significantly reduced the adult fecundity and longevity of F0H. axyridis and decreased the survival time and survival rate of the F1 generation. Sublethal concentrations of afidopyropen prolonged the developmental time of 4th instar larvae in the F0 generation. Additionally, acetamiprid and afidopyropen treatments significantly decreased the predation of H. axyridis. Furthermore, transcriptome sequencing analysis revealed that several P450 and UGT genes expressed differently when H. axyridis were exposed to sublethal concentrations of acetamiprid and afidopyropen, suggesting that the differential expression of detoxifying genes might be involved in the response and detoxification metabolism of acetamiprid and afidopyropen in H. axyridis. Our findings demonstrate that sublethal concentrations of acetamiprid adversely influences the development and predation of H. axyridis, while afidopyropen has limited effects on H. axyridis. These results are helpful for protecting and utilizing natural enemies and guiding the scientific use of pesticides in the field.

Characterization, expression, and functional analysis of TRPV genes in cotton aphid, Aphis gossypii Glover.

Transient receptor potential vanilloid (TRPV) channels have been found to be the molecular target of afidopyropen, a novel insecticide that is highly effective in controlling Aphis gossypii Glover in the field. However, the TRPV genes of A. gossypii has not yet been characterized. In this study, two TRPV genes of A. gossypii (AgNan and AgIav) were cloned and their expression levels were determined by quantitative real-time PCR (RT-qPCR). The deduced amino acids of AgNan and AgIav contain all conserved domains of TRPV and share very high amino acid identity with other insect TRPVs. AgNan and AgIav expressed in all developmental stages and their expression can be induced by afidopyropen in a dose- and time-dependent manner. Moreover, we found that silencing of AgNan and AgIav by RNA interference resulted in a significant mortality increase of adult A. gossypii compared to the control, which was even higher than 93 % at five days after feeding with dsAgIav, suggesting that knockdown of AgNan and AgIav have great effects on the survival of A. gossypii. The results of this study would be helpful for determining the reasonable use of afidopyropen in the integrated pest management programs of A. gossypii and provide useful information for further functional study of TRPVs in insects.

Identification and expression analysis of G protein-coupled receptors in the cotton aphid, Aphis gossypii Glover.

G protein-coupled receptors play important roles in mediating signal transformation and physiological processes. As a new type of insecticide target, GPCRs have attracted much attention in recent years. However, GPCRs have not yet been identified in Aphis gossypii. In the present study, a total of 87 GPCRs were identified from A. gossypii, including 65 Family A, 12 Family B, 7 Family C, and 3 Family F receptors. Most of the GPCRs in A. gossypii showed considerable sequence identity, and all of them have conserved transformmembrane domains. Newly identified GPCR genes were differentially expressed in different developmental stages and tissues. Moreover, we found that 34 GPCR genes were highly overexpressed in a sulfoxaflor-resistant strain, 4 and 10 of them were highly induced by LC15 and LC50 of sulfoxaflor, respectively. Furthermore, silencing of two highly overexpressed GPCRs by RNAi indicated that suppression the expression of AgoGPCR48 and AgoGPCR53 significantly increased the susceptibility of A. gossypii to sulfoxaflor, suggesting that these GPCR genes may be associated with sulfoxaflor resistance in A. gossypii. Our results imply that the overexpression of GPCR genes contribute to the sulfoxaflor resistance development in A. gossypii and provide useful targets for developing novel insecticides to manage this pest.

Effects of lufenuron treatments on the growth and development of Spodoptera frugiperda (Lepidoptera: Noctuidae).

Lufenuron is an effective benzoylurea insecticide that inhibits the synthesis of chitin and regulates the growth of insects. However, little is known about the effects of lufenuron treatment on the development of Spodoptera frugiperda (J. E. Smith). In this study, we assessed the toxicity of lufenuron on S. frugiperda and evaluated the effects of lufenuron treatment on the growth and development of S. frugiperda. The results showed that lufenuron exhibits high insecticidal activity against S. frugiperda, with the LC50 value of 0.99 mg L-1. Lufenuron treatments can significantly prolong the larval developmental duration and reduce the rates of pupation and emergence. To further explore the underlying mechanism of this observation, the expression profiles of the chitin synthase gene (SfCHS) and chitinase gene (SfCHT), two key enzyme genes involved in the molting of S. frugiperda, were determined after exposure to lufenuron for 96 h. The results of qRT-PCR demonstrated that lufenuron treatments can significantly reduce the expression of SfCHT, while the expression of SfCHS remained relatively stable. Furthermore, we found that lufenuron strongly interacted with chitinase (SfCHT) (-10.8 kcal/mol) and chitin synthase (SfCHS) (R1: -9.7 kcal/mol; R2: -10.2 kcal/mol). Our results indicated that lufenuron has significant effects on the development of S. frugiperda that might be attributed to the differential expression of SfCHT and SfCHS.

Regulation of COL1A2, AKT3 genes, and related signaling pathway in the pathology of congenital talipes equinovarus.

Congenital talipes equinovarus (CTEV) is one of the most common congenital limb defects in children, which is a multifactorial and complex disease that associates with many unknown genetic, social-demographic, and environmental risk factors. Emerging evidence proved that gene expression or mutation might play an important role in the occurrence and development of CTEV. However, the underlying reasons and involved mechanisms are still not clear. Herein, to probe the potential genes and related signaling pathways involved in CTEV, we first identified the differentially expressed genes (DEGs) by mRNA sequencing in pediatric patients with CTEV compared with normal children. The gene of COL1A2 was upregulated, and AKT3 was downregulated at the transcriptional level. Western blot and quantitative polymerase chain reaction (qRT-PCR) results also showed that the expression of COL1A2 in CTEV was enhanced, and the AKT3 was decreased. Furthermore, the COL1A2 Knock-in (+COL1A2) and AKT3 Knock-out (-AKT3) transgenic mice were used to verify the effects of these two genes in the CTEV, and the results of which showed that both COL1A2 and AKT3 were closely related to the CTEV. We also investigated the effect of the PI3K-AKT3 signaling pathway in CTEV by measuring the relative expression of several key genes using Western blot and qRT-PCR. In line with the Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis data, the PI3K-AKT3 signaling pathway might play a potentially important role in the regulation of pathological changes of CTEV. This study will provide new ideas for the mechanism investigation and prenatal diagnosis of CTEV.

Magnetic Resonance Imaging of Clubfoot Treated With the Ponseti Method: A Short-Term Outcome Study.

Objective: To quantitatively evaluate the effectiveness of the Ponseti method for the correction of clubfoot, we decided to use magnetic resonance imaging (MRI) to evaluate changes in the tarsal bone relationship. Methods: This is a retrospective study of fifteen children with clubfeet who were treated with the Ponseti method. MRI studies were obtained using a 3.0T Machine (GE Healthcare, United States). T1-weighted and T2-weighted images were acquired in the standard anatomic sagittal, transverse, and coronal planes. For the measurement, the best slice that clearly demonstrated the anatomy was chosen. Sagittal talocalcaneal angle, sagittal tibiocalcaneal angle, coronal tibiocalcaneal angle, transverse talar neck angle, transverse talonavicular angle, and transverse talocalcaneal angle were measured. The eighteen corrected clubfeet were compared with the twelve unilateral normal feet at clinical and radiological levels using a Pirani scoring system and MRI, respectively. Results: In total, 15 cases (twelve boys and three girls) with clubfeet were examined by using MRI. Twelve cases had unilateral and three had bilateral involvement (eleven left clubfeet and seven right clubfeet), giving a total of eighteen clubfeet when compared with twelve normal feet. The mean age of patients at examination was 47.7 months (8-96 months). The recovery of the corrected clubfoot in these patients met the goals of Ponseti treatment (functional, normal looking, pain-free, and plantigrade foot). Before Ponseti treatment, the mean Pirani score of clubfoot was 5.5 (5-6). During this follow-up, the Pirani score was 0.07 (0-0.05). The results of the MRI indicated that only the transverse talonavicular angle showed a significant difference between the treated clubfeet and the normal feet (p < 0.001). One case had dorsal talonavicular subluxation in the sagittal plane and had the lateral subluxation of the navicular in the transverse plane, which has never been reported in previous studies. Conclusion: Although the appearance and function of clubfoot were recovered well after the Ponseti method, the results of MRI indicated that the Ponseti method successfully corrected the varus, cavus, and equinus deformities and incompletely corrected the adduction deformity regarding transverse talonavicular angle. At the same time, the Ponseti method may cause dorsal talonavicular subluxation in the sagittal plane and lateral subluxation of the navicular in the transverse plane on MRI.

Expression of PAPP-A2 and IGF Pathway-Related Proteins in the Hip Joint of Normal Rat and Those with Developmental Dysplasia of the Hip.

Developmental dysplasia of the hip (DDH) is one of the major causes of child disability and early osteoarthritis. Genetic factors play an important role, but which still remain unclear. Pregnancy-associated plasma protein-A2 (PAPP-A2), a special hydrolase of insulin-like growth factor binding protein-5 (IGFBP-5), has been confirmed to be associated with DDH by previous studies. The aim of this study was firstly, to investigate the expression of PAPP-A2 and insulin-like growth factor (IGF) pathway-related proteins in normal rat's hip joints; secondly, to compare the variations of those proteins between DDH model rats and normal ones. The DDH model was established by swaddling the rat's hind legs in hip adduction and extension position. The hip joints were collected for expression study of fetal rats, normal newborn rats, and DDH model rats. Positive expression of PAPP-A2 and IGF pathway-related proteins was observed in all the hip joints of growing-stage rats. Ultimately, IGF1 was downregulated; insulin-like growth factor 1 receptor (IGF1R) showed an opposite trend in DDH rats when compared with normal group. The PAPP-A2 and IGF pathway-associated proteins may also be involved in the development of the rat's hip joint, which bring the foundation for further revealing the pathogenic mechanism of DDH.