Daphnis nerii can severely damage pine forests worldwide. Daphnis nerii cypovirus-23 (DnCPV-23) is an important viral pathogen for controlling D. nerii. However, the mechanism underlying DnCPV-23 cell entry has not been elucidated. In this study, we determined that VP3 mediates the binding of DnCPV-23 to host brush border membrane vesicles. Far-Western blotting and mass spectrometry results revealed that a Manduca sexta alpha-glucosidase (MsAGL) can interact with VP3. The interaction between MsAGL and VP3 was verified by co-immunoprecipitation and glutathione S-transferase pull-down assays. Notably, MsAGL influenced DnCPV-23 entry into host cells, including attachment and the subsequent internalization of the virus. Furthermore, MsAGL inhibited DnCPV-23 infections of M. sexta cells and D. nerii larvae. In summary, we confirmed that VP3 of DnCPV-23 mediates cell entry, while also identifying MsAGL as an entry factor for DnCPV-23. The study findings provide useful insights relevant for further elucidating the cell entry mechanisms of cypoviruses.

Sweet potato leaf worm (Aedia leucomelas; Lepidoptera: Noctuidae) adversely affects sweet potato quality and yield. We collected infected A. leucomelas larvae with typical symptoms of baculovirus infection in Fuzhou, China in 2023 and then isolated a baculovirus, which we named Aedia leucomelas nucleopolyhedrovirus (isolate Fuzhou;AeleNPV-FZ). Helicoverpa armigera and Spodoptera exigua larvae were also susceptible to AeleNPV-FZ in laboratory assays, suggesting that AeleNPV-FZ may be useful for controlling multiple noctuid pests under field conditions. The AeleNPV-FZ genome was sequenced and de novo assembled. The genome was 135,409 bp in length with 146 open reading frames (ORFs) and 12 homologous regions (hrs) annotated for the sequence. Phylogenetic relationships inferred from concatenated core gene amino acid sequence alignment and pairwise Kimura two-parameter distances indicated that AeleNPV-FZ may be a novel member of genus Alphabaculovirus.

In the original publication [...].

Culex pipiens quinquefasciatus is a notorious vector transmitting severe diseases such as Zika virus and West Nile virus to humans worldwide. Vermistatin is a type of funicon-like compound and was first isolated from Penicillin vermiculatum in the 1970s. Vermistatin has shown promising activity against Cx. p. quinquefasciatus larvae in our previous research. Here, we conducted a transcriptomic analysis of Cx. p. quinquefasciatus larvae treated with a median lethal concentration of 28.13 mg/L vermistatin. Differential expression analysis identified 1055 vermistatin-responsive genes, with 477 downregulated and 578 upregulated. Gene Ontology annotation and enrichment analysis revealed the metabolic process to be the most significantly affected biological process, the membrane to be the most significantly affected cellular component, and catalytic activity to be the most significantly affected molecular function. Kyoto Encyclopedia of Genes and Genomes pathway analysis classified the differential expression genes into six major categories, with metabolism and organismal systems being the most enriched. Fifty-five pathways were significantly enriched, with the hematopoietic cell lineage, renin–angiotensin system, cholesterol metabolism, and peroxisome proliferator-activated receptor signaling pathways among the top altered pathways. Furthermore, 32 potential detoxification-related genes were differentially expressed, with 3 cytochrome P450s, 2 ABC transporters, and 1 UGT induced by vermistatin. This study provides insights into the molecular mechanisms of vermistatin’s action against Cx. p. quinquefasciatus larvae, highlighting potential targets for novel mosquito control strategies.

Group 2 innate lymphoid cells (ILC2s) produce the type 2 cytokines IL-5 and IL-13 and contribute to type 2 immune responses, such as allergic airway inflammation. However, specific drugs, especially traditional Chinese medicines, that target lung ILC2s have rarely been reported. Here, we demonstrate that triptolide ameliorates allergic airway inflammation by suppressing ILC2 activation. IL-33, which is produced mainly by epithelial cells, is the most powerful cytokine for activating ILC2s. Triptolide-treated ILC2s were found to be functionally impaired in response to interleukin (IL)-33 challenge. RNA-seq analysis revealed that triptolide impaired ILC2 function through inflammation-related signalling pathways. ILC2-related genes were up- and down-regulated under the treatment with TPL such as Adrb2, Nmur1, tnfsf11, IL-5, IL-13, IL-9 and so on. Interestingly, we observed not only preventive but also therapeutic effects of triptolide on allergic airway inflammation, indicating that triptolide may serve as a promising traditional Chinese medicine for the treatment of allergic airway inflammation by targeting ILC2s.

Daphnis nerii cypovirus-23 (DnCPV-23) is a new type of cypovirus that has a lethal effect on many species of Sphingidae pests. DnCPV-23 can replicate in Spodoptera frugiperda Sf9 cells, but the replication characteristics of the virus in this cell line are still unclear. To determine the replication characteristics of DnCPV-23 in Sf9 cells, uninfected Sf9 cells and Sf9 cells at 24 and 72 h after DnCPV-23 infection were collected for transcriptome analysis. Compared to uninfected Sf9 cells, a total of 188 and 595 differentially expressed genes (DEGs) were identified in Sf9 cells collected at 24 hpi and 72 h, respectively. KEGG analyses revealed that 139 common DEGs in two treatment groups were related to nutrition and energy metabolism-related processes, cell membrane integrity and function-related pathways, detoxification-related pathways, growth and development-related pathways, and so on. We speculated that these cellular processes might be manipulated by viruses to promote replication. This study provides an important basis for further in-depth research on the mechanism of interaction between viruses and hosts. It provides additional basic information for the future exploitation of DnCPV-23 as a biological insecticide.

Allergic airway inflammation is a universal airway disease induced by inhaling allergens. Published data show that RNF128, an E3 ligase, promotes Th2 activation in the OVA-induced asthma model. Recent advances have shown that group 2 innate lymphoid cells (ILC2s) produce the cytokines IL-5 and IL-13 to mediate type 2 immune response. However, whether RNF128 regulates ILC2-dependent allergic lung inflammation remains unclear. In this study, we observed greater expression of the E3 ligase RNF128 in ILC2s than in other immune cells. RNF128 deficiency caused a selective increase in the number of peripheral mature ILC2s, and mice with RNF128 deficiency were more susceptible to Alternaria alternata (A. alternata) -induced allergic lung inflammation. Furthermore, RNF128 deficiency increased recruitment of eosinophils and levels of IL-5 and IL-13 in the bronchoalveolar lavage fluid. RNF128 effectively inhibited the expansion of ILC2s and the number of IL-5- and IL-13-producing ILC2s. Specially, RNF128 deficiency promoted the expression of the interleukin-33 (IL-33) receptor ST2 in A. alternata-induced allergic lung inflammation. Above all, our study demonstrated that RNF128 played a key role in A. alternata-induced ILC2 activation and type 2 immune response, suggesting that RNF128 may be an effective therapeutic target for allergic lung inflammation initiated by ILC2s.

Macrophages play an important role in the development of life-threatening sepsis, which is characterized by multiorgan dysfunction, through their ability to produce inflammatory cytokines. Carvacrol is a phenolic compound that has been confirmed to possess strong anti‑inflammatory activity. In this study, we mainly investigated the effect of carvacrol on lipopolysaccharide (LPS)-induced macrophage proinflammatory responses and endotoxic shock. The results showed that carvacrol significantly reduced mouse body weight loss and ameliorated pathological damage to the liver, lung, and heart under LPS-induced sepsis. Carvacrol attenuated inflammatory responses by inhibiting the LPS-induced production of inflammatory cytokine interleukin-6 (IL-6) in vivo and in vitro. Mechanistically, carvacrol inhibited IL-6 production mainly through the ERK1/2 signalling pathway in macrophages. Furthermore, carvacrol improved the survival of septic mice. This study sheds light on the role of carvacrol in the pathogenesis of LPS-induced sepsis, and thus, its potential in treating sepsis patients may be considered.

Daphnis nerii(Linnaeus)is a notorious pest that has a wide geographic distribution.In this study,we employed a mechanical crushing method to establish primary cultures of Daphnis nerii eggs and determined Grace's insect medium as the most optimal culture medium.Primary culture of Daphnis nerii embryonic cells lasted up to 12 months,and various forms of primary cells were differ-entiated in the primary culture.Our result provides crucial experimental basis for establishing Daph-nis nerii cell lines in future research.

Clanis bilineata Walker, soybean hawkmoth, belongs to the subfamily Ambulicinae (Sphingidae, Lepidoptera) and is an edible insect that usually grows on soybean leaves. In this study, we isolated a new cypovirus from naturally diseased Clanis bilineata larvae (named CbCPV), scanned its structure, sequenced its genome, and studied its phylogenetic relationship to other cypoviruses. Microscopy showed that CbCPV polyhedral occlusion bodies were about 1.878 μm on average and contained many virions in the ultrathin sections. The complete genome sequence of CbCPV is 22,812 bp comprising 10 segmented double-stranded RNAs. Apart from segment 1 containing one open reading frame (ORF) and one sub-ORF, the other nine segments all contain one open reading frame and encoded one putative protein. The non-coding regions contained conserved sequences at 5’ termini (AGUCAAA) and 3’ termini (AGC), except segment 4 containing a different 5’ termini (AUGUUUA). The whole sequence of the polyhedrin gene in CbCPV contained 892 nucleotides, encoding a protein of 246 amino acids. Based on amino acid sequences of polyhedrin or RNA dependent RNA polymerase (RdRp), the phylogenetic analysis indicated that CbCPV was closely related to DnCPV-23. The putative function of all segments differed from each other, but the most closely related species of segments were DnCPV-23 with 98.2–99.8

•37 compounds were successfully identified from Origanum vulgare essential oil.•O. vulgare EO were toxic to Aedes albopictus and Culex pipiens quinquefasciatus.•O. vulgare EO’s major compounds thymol and carvacrol were toxic to mosquito.•Piperonyl butoxide improved the toxicity of O. vulgare EO, thymol and carvacrol.

Asthma is an inflammatory disease of the airway that is prone to recurring episodes. Patients have clinical symptoms such as airway hyperresponsiveness (AHR), airway remodeling, increased mucus secretion, and airflow limitation. The incidence of asthma is affected by many factors such as genetic factors and environmental factors. Asthma is mainly divided into allergic asthma and nonallergic asthma. T helper 2 (Th2) cells have long been considered to play a key role in regulating the adaptive immune response in airway inflammation. Allergic asthma is also known as Th2 asthma. With the discovery of non Th2 asthma and innate lymphocytes (ILCs), the function of ILCs in asthma has gradually become the research focus. Recent research demonstrates that ILC2s represent a crucial source of cytokines and could regulate the adaptive immune response in both allergic asthma and non-allergic asthma. In this review, we summarize the latest advances in the function of ILCs in asthma.

从自然死亡的雀纹天蛾幼虫分离到一株雀纹天蛾核型多角体病毒.通过扫描及切片透射电镜发现,该病毒为单粒包埋型核型多角体病毒,命名为ThjaSNPV(Theretra japonica single nucleopolyhedrovirus).病毒全基因组重测序后拼接显示,该病毒基因组全长134 899 bp,GC含量37.28％,与豆天蛾单粒包埋型核型多角体病毒株DZ1基因组序列相似性高达96.24％.ThjaSNPV含有131个开放阅读框(ORF)其中55个为正链基因,76个为负链基因,与宿主同为天蛾科的豆天蛾单粒包埋型核型多角体基因组相比,雀纹天蛾单粒包埋型核型多角体病毒新注释到7个基因:chitin-binding protein(ORF9),ODV-E18(ORF11),-lef-11(ORF27),hypothetical protein(ORF41),lef-10(ORF42),pif-6(ORF66),P6.9(ORF88).ThjaSNPV 的 DNA 光裂酶基因(DNA photolyase,ORF53)中不具有豆天蛾NPV中的1 bp碱基的缺失,只编码一个大的完整DNA裂合酶.38个串联的核心基因的进化分析显示雀纹天蛾NPV与Alphabaculovirus group Ⅱ类群聚类在一起,且和豆天蛾NPV最为相似.

Abstract Polycystic kidney disease (PKD) is a genetic disorder characterized by uncontrolled proliferation of renal cells, with the consequent formation of cysts and loss of renal function. Matrine has the effect of regulating autophagy, and is considered to regulate inflammatory responses and cyst formation. Therefore, in this study we focused on the pathological mechanism of matrine-regulated autophagy in polycystic kidney disease, and identified some autophagy-regulated genes. We also performed transcriptome sequencing of matrine-treated mouse renal epithelial cells (IMCD3). The pathway analysis results showed that signal transduction, including adrenergic signaling in cardiomyocytes, Hippo signaling pathway, and calcium signaling pathway, which are closely related to autophagy, comprises the main pathological changes of IMCD3 cells treated with matrine. These results indicate that exaggerated autophagy participates in the pathological process of polycystic kidney disease, and may provide new insight for further basic research on PKD.

草原毛虫(Gynaephora ruergensis Chou et yin)是我国高原牧草中的一种有害害虫.草原毛虫核型多角体病毒新分离株(EupsNPV-Gr)具有典型的杆状病毒特征,呈单粒包埋型病毒粒子,ODV颗粒呈不规则的多边形,直径为1.0μm～1.35μm.EupsNPV-Gr能有效杀灭草原毛虫幼虫,它对草原毛虫的半致死浓度LC50为4×104 PIB/mL,是一种理想的生物防治剂.本研究对第一个EupsNPV-Gr基因组进行了测序和鉴定.EupsNPV-Gr基因组全长140684bp,包含134个开放阅读框.它与茶毛虫核型多角体病毒(EupsNPV)关系最为密切,序列同源性高达99％.系统进化分析表明,EupsNPV-Gr属于杆状病毒科alpha杆状病毒属,根据公认的种属划分标准,EupsNPV-Gr是EupsNPV的一个株系.新报道的基因组为研究该物种毒力的分子机制提供了基础.

Penicillium oxalicum T1能在9 d内将天然水稻秸秆降解成汤液.从第2天开始pH值急剧下降,第4―6天pH值维持在4左右,到第7天pH值下降到3左右,降解率达64.7％.扫描电镜观察到随着降解时间的延长,稻草秸秆的蜡质和半纤维素层降解脱落,露出纤维素和硅结构,秸秆的表面裂解的情况越来越严重.采用气相色谱-四级杆质谱联用仪检测到汤液中有2-苯乙醇,采用气相色谱检测2-苯乙醇含量为1.0 g/L.

本实验主要研究20亿PIB/mL松毛虫质型多角体病毒悬浮剂防治马尾松毛虫田间药效.实验结果表明:松毛虫质型多角体病毒的致死速度与其剂量有关,在500倍液剂量处理区,在1周内的死亡率达到67.6％,各试剂量区均有明显的持效性.第15天的防效效果为83.5％ ～90.4％;第30天的防效为85.6％ ～95％;松毛虫质型多角体病毒1000倍液剂量与500倍液剂量区与Bt区防治效果差异显著.

根据时代需要,利用环介导等温扩增技术(loop-mediated isothermal amplifcation,LAMP)对新冠病毒进行快速检测,以研发出新冠病毒检测试剂盒.通过NCBI公布的新冠基因组序列分析,获得新冠病毒特异性N蛋白基因的序列;根据该序列信息,在线设计的几对环介导等温扩增引物以供筛选,并利用新冠病毒全基因的反转录质粒,同时通过大量的实验尝试,得到较为稳定的环介导等温扩增体系,再加入具有逆转录活性的Bst 3.0 DNA聚合酶,使体系可扩增RNA模板,最后通过加入适量染料,完成反应结果的可视化.通过不断的实验优化,得到了较为稳定的扩增体系如下:2×Lamp Master Mix 12.5μL,Bst 3.0 DNA聚合酶0.5μL,内引物FIP、BIP(10μM)各2μL、外引物F3、B3(10μM)各0.5μL、模板1μL、ddH2 O 7μL,该体系在65℃下水浴加热1 h即可,同时在体系中加入MnCl2溶液(15 mM)1μL、钙黄绿素(500μM)3μL,使反应结果肉眼可断,加入模板的体系变为绿色,而未加模板的体系为棕黄色.结论:本实验研究的可视化LAMP检测新冠病毒的方法可用于对广大人民群众进行新冠病毒核酸检测.

新型夹竹桃天蛾质型多角体病毒(Daphnis nerii cypovirus-23,DnCPV-23)可杀灭多种天蛾科昆虫,具备开发新型昆虫病毒杀虫剂的潜力.目前,主要利用夹竹桃天蛾(Daphnis nerii)对该病毒进行研究,DnCPV-23在细胞中的复制情况尚不清楚.本研究将DnCPV-23病毒粒子接种至草地贪夜蛾(Spodoptera frugiperda)细胞系Sf9中,分析该病毒在细胞中的复制情况.将病毒感染的细胞置于27℃条件下进行培养,利用光学显微镜观察病毒对Sf9细胞状态的影响;分别提取第1、3和7 d感染细胞的RNA,以其为模板进行反转录获得cDNA,利用qRT-PCR分析DnCPV-23 RNA水平;利用qRT-PCR方法分别对细胞培养上清液和病毒连续传代的细胞进行研究.结果显示,DnCPV-23能较好地适应Sf9细胞且成功感染Sf9细胞,并产生细胞病变效应(cytopathic effect,CPE);在细胞培养上清液中能够检测到病毒片段;病毒能够在Sf9细胞上进行传代.该研究为深入研究DnCPV-23入侵、转录机制、评价病毒毒力提供了良好的细胞模型,同时为新型天蛾科害虫病毒杀虫剂的开发提供一定的参考和依据.

Background Daphnis nerii cypovirus-23 (DnCPV-23) is a new type of cypovirus and has a lethal effect on the oleander hawk moth, Daphnis nerii which feeds on leave of Oleander and Catharanthus et al. After DnCPV-23 infection, the change of Daphnis nerii responses has not been reported. Methods To better understand the pathogenic mechanism of DnCPV-23 infection, 3rd-instar Daphnis nerii larvae were orally infected with DnCPV-23 occlusion bodies and the transcriptional responses of the Daphnis nerii midgut were analyzed 72 h post-infection using RNA-seq. Results The results showed that 1979 differentially expressed Daphnis nerii transcripts in the infected midgut had been identified. KEGG analysis showed that protein digestion and absorption, Toll and Imd signaling pathway were down-regulated. Based on the result, we speculated that food digestion and absorption in insect midgut might be impaired after virus infection. In addition, the down-regulation of the immune response may make D. nerii more susceptible to bacterial infections. Glycerophospholipid metabolism and xenobiotics metabolism were up-regulated. These two types of pathways may affect the viral replication and xenobiotic detoxification of insect, respectively. Conclusion These results may facilitate a better understanding of the changes in Daphnis nerii metabolism during cypovirus infection and serve as a basis for future research on the molecular mechanism of DnCPV-23 invasion.