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Neonicotinoids have recently been banned in the European Union due to concerns about their environmental safety. Thiacloprid is a neonicotinoid, and is frequently used in agricultural pest control. However, the lack of quantitative methods for assessing its multiple metabolites has made it difficult to perform a comprehensive safety assessment. Therefore, we established an analytical method for effectively determining thiacloprid and its five metabolites in vegetables and flowers using QuEChERS-HPLC-MS/MS. The samples were extracted with acidified acetonitrile (containing 0.5% acetic acid) and were cleaned with graphitised carbon black (GCB) and octadecylsilane (C18) before the residue analysis. This method provided linearity ranging from 2–2000 μg kg−1, and its linear regression coefficients exceeded 0.995. Limits of Quantitation (LOQs) are achieved at 2 μg kg−1， and Limits of Detection (LODs) ranged from 0.09–0.80 μg kg−1. The method was validated on seven matrices spiked at 2, 200, and 2000 μg kg−1. Average recoveries ranged from 77–119%, with an intra-day Relative Standard Deviation (RSDr) of 1.5–13.4% and an inter-day Relative Standard Deviation (RSDR) of 0.9–14.4%. This method was used on cucumbers sprayed with thiacloprid and 19 types of flower samples collected from the field, after which thiacloprid and several of its metabolites were detected.

Seeds play a critical role in the production of American ginseng. Seeds are also one of the most important media for the long-distant dissemination and the crucial way for pathogen survival. Figuring out the pathogens carried by seeds is the basis for effective management of seed-borne diseases. In this paper, we tested the fungi carried by the seeds of American ginseng from main production areas of China using incubation and highly throughput sequencing methods. The seed-carried rates of fungi in Liuba, Fusong, Rongcheng, and Wendeng were 100%, 93.8%, 75.2%, and 45.7%, respectively. Sixty-seven fungal species, which belonged to twenty-eight genera, were isolated from the seeds. Eleven pathogens were identified from the seed samples. Among the pathogens, Fusarium spp., were found in all of the seed samples. The relative abundance of Fusarium spp. in the kernel was higher than that in the shell. Alpha index showed that the fungal diversity between seed shell and kernel differed significantly. Non-metric multidimensional scaling analysis revealed that the samples from different provinces and between seed shell and kernel were distinctly separated. The inhibition rates of four fungicides to seed-carried fungi of American ginseng were 71.83% for Tebuconazole SC, 46.67% for Azoxystrobin SC, 46.08% for Fludioxonil WP, and 11.11% for Phenamacril SC. Fludioxonil, a conventional seed treatment agent, showed a low inhibitory effect on seed-carried fungi of American ginseng.

Plants have evolved complex signaling networks that enable them to adapt to adverse environmental conditions. The dehydration-responsive element-binding (DREB) transcription factors are important for plant responses to abiotic stresses. In this study, a new member of the AP2/ERF transcription factor gene family, PcDREB2A, was cloned and characterized from Polygonum cuspidatum, a traditional Chinese medicinal herb. PcDREB2A, which includes a typical AP2 domain, was clustered in the A-2 subgroup of the DREB subfamily. At the seedling stage, PcDREB2A expression was induced by cold, salt, and drought stresses. A yeast one-hybrid assay and an analysis of transiently transformed tobacco revealed that PcDREB2A can specifically bind to the DRE motif and transactivate reporter gene expression. Following 200 and 250 mM mannitol treatments, the PcDREB2A-overexpressing Arabidopsis thaliana lines had longer roots and a significantly higher fresh weight than the wild-type plants. Furthermore, under drought stress conditions, the PcDREB2A-overexpressing A. thaliana plants accumulated less malondialdehyde than the control plants. These results indicate that PcDREB2A encodes a novel DREB transcription factor in P. cuspidatum. Furthermore, the data generated in this study may be useful for researchers and breeders interested in genetically engineering plants to increase drought tolerance without inhibiting growth.

Agricultural soils and microplastics (MPs) are hotspots for antibiotic resistance genes (ARGs). Plastic mulch is the most important source of MPs in agricultural soil. ARGs, mobile genetic elements (MGEs), and their host profiles in long-term mulch MP-exposed soils remain unclear. In the present study, metagenomics was used to investigate the distribution patterns of ARGs and MGEs in eight Chinese provinces with a long history of plastic mulch use. A total of 204 subtypes of ARGs and thousands of MGEs (14 integrons, 28 insertions, and 2993 plasmids) were identified. A similar diversity of ARGs was found among MPs film-contaminated sites. The types of ARGs with a high abundance were more concentrated, and multidrug resistance genes were the dominant ARGs. Soils from regions with a longer history of plastic film use (such as Xinjiang province) had a higher abundance of ARGs and MGEs. The distribution of ARGs and MGEs exhibited a modular network distribution pattern. A total of 27 ARG subtypes and 29 MGEs showed co-occurrence network relationships. More than 10 common hosts of ARGs and MGEs, such as Pseudomonas, were found, and their abundances were highest in three provinces, including Xinjiang. This study may help elucidate the impact mechanism of long-term MP residues on the occurrence and spread of ARGs in soil.

Lufenuron (LUF) and Methoxyfenozide (MET) as Insect Growth Regulators (IGRs) contribute to the current control of the catastrophic crop pest, Spodoptera exigua (Lepidoptera, Noctuidae). Yet S. exigua has evolved resistance to LUF and MET, which is possibly mediated by cytochrome P450 monooxygenases (P450s), particularly from the CYP3 clade family, as it plays a key role in the detoxification of insecticides. However, a mixture of LUF and MET (MML) (optimal ratio: 6:4) remains highly insecticidal. Here, we analysed the response of S. exigua to sublethal concentrations of LUF, MET, and MML via transcriptomics. Twelve differentially expressed genes (DEGs) encoding CYP3 clade members were observed in transcriptomes and CYP9A9 was significantly upregulated after treatment with LUF, MET, and MML. Further, CYP9A9 was most highly expressed in the midgut of L4 S. exigua larvae. RNAi-mediated knockdown of CYP9A9 reduced the activity of CYP450 and increased the susceptibility of S. exigua larvae to LUF, MET, and MML. Thus, CYP9A9 plays a key role in the detoxification of LUF, MET, and MML in S. exigua. These findings provide new insights into insecticidal actions of IGRs, which can be applied to the establishment of novel pest management strategies.

Gonadal hormone is essential for the health of postmenopausal women, however, few studies have focused on the epidemiological distribution of gonadal hormones in postmenopausal women in very late postmenopausal women. This study aims to investigate and analyze the differences of serum gonadal hormone content and its influential factors among female centenarians in Hainan, China.The questionnaire and physical examination data of 741 female centenarians and 401 elderly females in Hainan Province were collected, and venous blood samples were taken to detect the indexes of lipid metabolism, bone metabolism, and gonadal hormone. The differences of gonadal hormones and relavant factors in female centenarians were analyzed and compared.The serum levels of estradiol and progesterone of female centenarians were significantly higher than those of the elderly females (bothFor the extremely late postmenopausal women (such as centenarians), there may be characteristic expressions of gonadal hormones, especially estradiol. There is an unprotective correlation of serum estradiol with lipid metabolism index and bone metabolism index in female centenarians, so it is necessary to evaluate the estrogen content and the use of estrogen therapy in postmenopausal women.

Spodoptera exigua (Lepidoptera, Noctuidae) has been responsible for causing considerable and widespread agricultural losses worldwide. Owing to strong selective pressure, S. exigua showed increased resistance to Lufenuron (LUF). Consequently, RNA interference (RNAi)-based insecticides had more benefits than chemical insecticides. Therefore, to enhance the insecticidal activity of LUF to S. exigua, in the present study, we aimed to elucidate the impact of double-stranded RNAs (dsRNAs) on S. exigua larval susceptibility to LUF. First, the transcriptome of S. exigua was sequenced following the treatment with LUF. By comparing the upregulated and downregulated GO enrichment, chitin binding and chitin metabolic processes were the significantly enriched pathways. According to transcriptome sequencing, 8 genes associated with chitin biosynthesis, 8 chitin degradation genes, and 17 cuticle protein genes were obtained. UDP-N-acetylglucosamine pyrophosphorylase (UAP) and Chitin synthase A (CHSA) showed significantly downregulated expression after treatment with different sublethal doses of LUF. Downregulation of UAP increased mortality from 31.97% to 47.91% when the larvae were exposed to LUF. A significant increase in the mortality of S. exigua from 30.63% to 50.19% was observed following LUF administration after dsCHSA. In addition, the expression analysis of genes associated with chitin biosynthesis was significantly changed after LUF treatment, dsRNAs-RNAi, and their combination (LUF-dsRNAs). Significant differences were observed in the chitin content between the control group at 72 h after treatments. Results of the present study can help further elucidate the understanding of the combined effects of RNAi and LUF on S. exigua. Additionally, this research provides a suitable foundation for future studies with the aim to develop an efficient method of delivery for large-scale pest control in the fields.

Correction for ‘Distinctive organized molecular assemble of MoS2, MOF and Co3O4, for efficient dye-sensitized photocatalytic H2 evolution’ by Kai Fan et al., Catal. Sci. Technol., 2018, 8, 2352–2363, https://doi.org/10.1039/C8CY00380G.

Unmanned aerial vehicles (UAVs) are widely used as the sprayers for low-volume pesticide application in recent years. Droplet distribution characteristics of UAV spraying in the cotton canopy have notable effect on the biological control efficacy of the targets and the defoliation efficiency of the harvest aids. In this work, the influences on droplet distribution in the cotton canopy with respect to the flight height, forward mode, and spraying volume were evaluated by conducting the field trials during two cotton growth stages in 2020, respectively. The first field trial was performed in the cotton flowering stage and the second one was conducted in the early boll development stage. Two typical UAVs equipped with a single-rotor and four-rotor, respectively, were adopted as the spraying platforms in this work. Droplet deposition obtained by water sensitive papers (WSPs) clipped on the cotton leaves was considered as the observing metric. All cotton leaves in the canopy were divided into three groups (i.e., upper, middle, and bottom layers) in both trials. Furthermore, the cotton canopy was divided as eight directions to assess the droplet distribution in the canopy from different directions. The results showed that the droplet deposition varied remarkable between the treatments and in the same canopy within a treatment. The upper layer obtained higher droplet deposition than those of the middle and bottom layers and plants P4 to P8 accessed more droplets than those of the remaining sampling plants in most treatments of both trials for the two UAVs. The upper layer droplet deposition of the four-rotor UAV treatments outperformed that of the single-rotor treatments under the same operating parameters. The forward modes rarely affected the droplet distribution of the four-rotor UAV treatments but significantly influenced that of the single-rotor UAV treatments. For the single-rotor UAV spraying with “head forward”, the droplet distribution of the treatment with a flight height of 2 m was more even than that of the 1 and 3 m in the first trial. Under the same flight height, droplet deposition of the treatments with a spraying volume of 22.5 L ha−1 was remarkably higher than that of the 12 L ha−1 for both forward modes in the second trial. “Tail forward” of the single-rotor UAV treatment had better penetration at a flight height of 2 m in both trials. Therefore, for the single-rotor UAV, under a flight height of 2 m and a spraying volume of 22.5 L ha−1, “tail forward” was recommended for applying pesticides to control targets at the lower canopy and “head forward” was a better choice for harvest aid application. Four-rotor UAV was a suitable adoption for the harvest aid application and controlling the targets of the upper canopy. The results also indicate that the systemic pesticides are recommended for UAV spraying due to its uneven droplet distribution uniformity in the whole cotton canopy.

Burkholderia pseudomallei is a Gram-negative soil-dwelling bacillus that causes melioidosis, a frequently fatal infectious disease, in tropical and subtropical regions. Previous studies have identified the overall genetic and evolutionary characteristics of B. pseudomallei on a global scale, including its origin and transmission routes. However, beyond its known hyperendemicity foci in northern Australia and Southeast Asia, the distribution and genetic characteristics of B. pseudomallei in most tropical regions remain poorly understood, including in southern China. Here, we sequenced the genomes of 122 B. pseudomallei strains collected from Hainan, an island in southern China, in 2002–2018, to investigate the population structure, relationships with global strains, local epidemiology, and virulence and antimicrobial-resistance factors. A phylogenetic analysis and hierarchical clustering divided the Hainan strains into nine phylogenic groups (PGs), 80 % of which were concentrated within five major groups (group 1: corresponding to minor sequence types [STs], 12.3 %; group 3: ST46 and ST50, 31.1 %; group 9: ST58, 13.1 %; group 11: ST55, 8.2 %; group 15: mainly ST658, 15.6%). A phylogenetic analysis that included global strains suggested that B. pseudomallei in Hainan originated from Southeast Asian countries, transmitted in multiple historical importation events. We also identified several mutual transmission events between Hainan and Southeast Asian countries in recent years, including three importation events from Thailand and Singapore to Hainan and three exportation events from Hainan to Singapore, Malaysia, and Taiwan island. A statistical analysis of the temporal distribution showed that the Hainan strains of groups 3, 9, and 15 have dominated the disease epidemic locally in the last 5 years. The spatial distribution of the Hainan strains demonstrated that some PGs are distributed in different cities on Hainan island, and by combining phylogenic and geographic distribution information, we detected 21 between-city transmission events, indicating its frequent local transmission. The detection of virulence factor genes showed that 56 % of the Hainan strains in group 1 encode a B. pseudomallei -specific adherence factor, boaB, confirming the specific pathogenic characteristics of the Hainan strains in group 1. An analysis of the antimicrobial-resistance potential of B. pseudomallei showed that various kinds of alterations were identified in clinically relevant antibiotic resistance factors, such as AmrR, PenA and PBP3, etc. Our results clarify the population structure, local epidemiology, and pathogenic characteristics of B. pseudomallei in Hainan, providing further insight into its regional and global transmission networks and improving our knowledge of its global phylogeography.

As microplastics became the focus of global attention, the hazards of plastic plasticizers (PAEs) have gradually attracted people's attention. Agricultural soil is one of its hardest hit areas. However, current research of its impact on soil ecology only stops at the microorganism itself, and there is still lack of conclusion on the impact of soil metabolism. To this end, three most common PAEs (Dimethyl phthalate: DMP, Dibutyl phthalate: DBP and Bis (2-ethylhexyl) phthalate: DEHP) were selected and based on high-throughput sequencing and metabolomics platforms, the influence of PAEs residues on soil metabolic functions were revealed for the first time. PAEs did not significantly changed the alpha diversity of soil bacteria in the short term, but changed their community structure and interfered with the complexity of community symbiosis network. Metabolomics indicated that exposure to DBP can significantly change the soil metabolite profile. A total of 172 differential metabolites were found, of which 100 were up-regulated and 72 were down-regulated. DBP treatment interfered with 43 metabolic pathways including basic metabolic processes. In particular, it interfered with the metabolism of residual steroids and promoted the metabolism of various plasticizers. In addition, through differential labeling and collinear analysis, some bacteria with the degradation potential of PAEs, such as Gordonia, were excavated.

Intracoronary application of nicorandil can effectively reduce the myocardial no‐reflow (MNR) after percutaneous coronary intervention (PCI). We sought to investigate the mechanisms of nicorandil in preventing MNR, besides that of dilating the coronary microvasculature. A total of 60 patients undergoing PCI were enrolled and randomly divided into a nicorandil group and a control group. Before PCI, 2 mg of nicorandil or an equal volume of normal saline was injected into the coronary artery. Blood samples were collected before, 24 hours and 1 week after PCI and inflammatory cytokines were tested. In the control group, the expression of pro‐inflammatory cytokines was significantly increased, while the anti‐inflammatory cytokines were decreased 24 hours after PCI. In contrast, these changes were reversed in the nicorandil group, indicating that nicorandil regulated the inflammatory response induced by PCI. Then, proteomic analysis was performed to further elucidate the potential mechanisms. A total of 53 differentially expressed proteins (DEPs) were found 24 hours after PCI in the control group, and the changes of these relevant genes were reversed in the nicorandil group. These DEPs were significantly enriched in the inflammatory pathways. In conclusion, the intracoronary application of nicorandil before PCI can regulate the inflammatory responses induced by PCI, which might be an important mechanism of nicorandil in preventing MNR.

Lung cancer patients with lymph node metastasis usually had short overall survival and occurred distant metastases at the early stage. However, some of these people did have more prolonged survival. The underlying reason is still unclear. In this study, we found a novel molecule, family with sequence similarity 136, member A gene (FAM136A). First, we performed immunohistochemistry for FAM136A in 177 lung carcinoma tissues. Second, we carried out in vitro studies by using A549 and PC-9. We detected FAM136A immunoreactivity in 79 out of 177 (44.6%) lung carcinoma tissues, and the FAM136A status was significantly associated with tumor T stage, lymph node metastasis, and the Tumor-Node-Metastasis (TNM) staging system in these cases. Importantly, it was significantly associated with the overall survival of the patients with lymph node metastasis, especially FAM136A positive patients, who had worse outcomes. Subsequent in vitro experiments revealed that the proliferation activity and migration property decreased both A549 and PC-9 lung carcinoma cells transfected with siRNA-FAM136A, and apoptosis reduced. Meanwhile, the expression of CDK4 and CDK6 decreased. FAM136A status would be a potent, worse prognostic factor in lung cancer patients with lymph node metastasis. It would play a vital role in the proliferation, apoptosis, and migration properties of A549 and PC-9. In the future, We will focus on the uncovered signal mechanism between FAM136A and lung cancer., Graphical Abstract Abbreviation: FAM136A: family with sequence similarity 136 member A; NSCLC: non-small-cell lung cancer; SCC: squamous cell carcinoma; IHC: immunohistochemistry; PBS: phosphate-buffered saline; CDK: cyclin-dependent kinase.

Background Cancer is a leading cause of death worldwide; folk anticancer medicinal plants have applied for cancer treatment. The Maytenus compound tablet as traditional Chinese compound medicine has been approved for alleviating hyperplasia of mammary glands, whether it can inhibit cancer still unknown. The study was to evaluate the anticancer activity of the Maytenus compound tablet. Methods MTS assay evaluated the anti-proliferation effect of the Maytenus compound on H226, A2058, 786O and HeLa cancer cells and two normal epithelial cell lines, namely, 16HBE and Hecate. Nude mouse xenograft tumor model using H226 and HeLa cells examined the drug’s anticancer effect in vivo. Western blot assay studied the possible mechanism. Results The Maytenus compound indicated obvious ability to against proliferation in four strains of cancer cells, particularly against H226 cells by an IC50 of 85.47±10.06 µg/mL and against HeLa cells by an IC50 of 128.74±17.46 µg/mL. However, it had a low cytotoxicity in human normal epithelial cell lines 16HBE with an IC50 of 4,555.86±25.21 µg/mL and Hecate with an IC50 of 833.56±181.88 µg/mL. The Maytenus compound at the 2.45 g/kg oral dosages inhibited the proliferation of H226 cells and HeLa cells in nude mouse with inhibitory rates of 36.06% and 26.45%, respectively, and no organ toxicity. The Maytenus compound could significantly downregulate the expression of pEGFR, pPI3K, pAKT, pGSK3β, β-catenin, and c-MYC and upregulate the protein expression of GSK3β. Conclusions The Maytenus compound has significant anticancer activities against human cancer H226 and HeLa cells both in vitro and in vivo, highlighting it may be an anticancer medicine.

Background: A rare member of metallo-β-lactamases genes, blaSIM-1, carried by a 316-kb plasmid designated pSIM-1-BJ01 was isolated from a clinical cephalosporins- and carbapenem-resistant Klebsiella pneumoniae 13624. This is the first sequence report of a transferable blaSIM-1-carrying conjugative plasmid isolated from K. pneumoniae. Purpose: The sequence analysis of pSIM-1-BJ01 will help us to identify genes responsible for conjugation, plasmid maintenance and drug resistance, to understand the evolution and control the dissemination of resistance plasmids. Patients and methods: K. pneumoniae 13624 was isolated from the urine specimen of a patient. Bacterial genomic DNA was sequenced with PacBio RSII platform. Results: Most of the pSIM-1-BJ01 backbone matches that of pRJA166a, which was isolated from a clinical hypervirulent K. pneumoniae ST23 strain at Shanghai, China, recently. The highly homologous backbones between the two plasmids imply the close relationship of evolution. Two different multidrug-resistant regions both carrying the class 1 integrons with different resistance genes have been assembled into the pSIM-1-BJ01. Besides, the other two resistance plasmids, pKP13624-1 carrying blaTEM-1 and blaCTX-M-15 and pKP13624-2 carrying blaCTX-M-14 and blaLAP-2 were also identified. Conclusion: The emergence of the blaSIM-1-carrying IncHI1B pSIM-1-BJ01 suggests the spread of blaSIM among Enterobacteriaceae is possible. We should pay more attention to supervise and control the dissemination of hypervirulent carbapenem-resistant K. pneumonia in public hospitals.

China has been piloting the catastrophic medical insurance (CMI) program since 2012 and rolled it out nationally in 2016 to reduce the incidence of catastrophic health expenditure among Chinese residents. Few studies have been conducted to determine its effect on healthcare expenditures, particularly among the elderly. The purpose of this study is to examine the effect of CMI on healthcare expenditures among China’s elderly population. The data for this study were derived from 4 waves of the Chinese Longitudinal Healthy Longevity Survey, which included 344 and 1199 individuals in the treatment and control groups, respectively. To examine the effect of CMI on healthcare expenditures among the elderly, we used difference-in-differences and fixed-effects models. Additionally, a heterogeneity analysis was used to examine the differences in the impact of CMI on different groups. Finally, we confirmed the robustness of the results using robustness and placebo tests. CMI increased total health and out-of-pocket expenditures significantly, as well as inpatient and corresponding out-of-pocket expenditures. The reassults of the heterogeneity analysis indicated that CMI had a greater impact on elderly residents of rural areas. Economic burden protection has been enhanced for low-income groups and patients with serious diseases over the last two years. Our research indicated that CMI can promote the use of inpatient medical services for the elderly to a certain extent. Targeted measures such as expanding the CMI compensation list, establishing a more precise compensation scheme, and specific diseases associated with high healthcare expenditures can be considered in the practice of CMI implementation.

Plastic pollution in the soil ecosystem is currently receiving worldwide attention. However, little is known whether the presence of microplastics (MPs) in soil will affect the environmental behavior of pesticide residues in soil. Here, the effect of the addition of new mulch MPs (New-MPs), aged mulch MPs (Aged-MPs) and biodegradable mulch MPs (BioD-MPs) on the adsorption and degradation behaviors of two pesticides (imidacloprid and flumioxazin) in soil was investigated. Three MPs slowed down rapid adsorption stage of pesticides in soil and delayed the time to reach adsorption equilibrium. Adsorption rates: Soil Soil + New-MPs Soil + Aged-MPs Soil + BioD-MPs. Three MPs enhanced the adsorption strength of the soil system for the two pesticides, and the aging treatment of the MPs enhanced this effect. Three MPs affected the degradation process of the two pesticides. New-MPs promoted the degradation of two pesticides imidacloprid and flumioxazin, and the degradation half-lives were shortened to 0.93 and 0.85 times, respectively; while Aged-Mps and BioD-MPs delayed the degradation process of two pesticides, and the degradation half-lives were extended to 1.64 times and 1.21 times, respectively. The effect was more significant with the increase of MPs and pesticides concentration. Pesticide polarity, surface structure and functional groups of MPs are potentially important reasons for the differences in adsorption and degradation of MPs-soil systems. Our findings provide a deep insight into understanding the mechanism of interaction between MPs and pesticide residues in soil environment.

Spodoptera exigua is a worldwide pest afflicting edible vegetables and has developed varying levels of resistance to insecticides. Methoxyfenozide (MET), an ecdysteroid agonist, is effective against lepidopteran pests such as S. exigua. However, the mechanism of MET to S. exigua remains unclear. In this study, we analyzed the expression patterns of genes related to the ecdysone signaling pathway in transcriptome data treated with sublethal doses of MET and analyzed how expression levels of key genes affect the toxicity of MET on S. exigua. Our results demonstrated that 2639 genes were up-regulated and 2512 genes were down-regulated in S. exigua treated with LC30 of MET. Of these, 15 genes were involved in the ecdysone signaling pathway. qPCR results demonstrated that ecdysone receptor A (EcRA) expression levels significantly increased in S. exigua when treated with different doses of MET, and that the RNAi-mediated silencing of EcRA significantly increased mortality to 55.43% at 72 h when L3 S. exigua larvae were exposed to MET at the LC30 dose. Additionally, knocking down EcRA suppressed the most genes expressed in the ecdysone signaling pathway. The combination of MET and dsEcRA affected the expression of E74 and enhanced the expression of TREA. These results demonstrate that the adverse effects of sublethal MET disturb the ecdysone signaling pathway in S. exigua, and EcRA is closely related to MET toxic effect. This study increases our collective understanding of the mechanisms of MET in insect pests.

Additional file 3: Figure S1 Length distribution of the assembled unigenes of Polygonum cuspidatum. Figure S2 Assessment of assemblies using Benchmarking Universal Single-Copy Orthologs (BUSCO). Figure S3 Overview of gene expression in Polygonum cuspidatum. (A) Boxplot analysis of gene expression profiles in the nine libraries. Boxes represent interquartile ranges; the line across the box represents the median; the plus sign in the box shows mean values; and hyphens over and under the boxes represent the maximum and minimum, respectively. (B) Principal component analysis (PCA) showing clustering pattern among different tissues based on global gene expression profiles. Group1: Root-1, Root-2, and Root-3; Group2: Stem-1, Stem-2, and Stem-3; Group3: Leaf-1, Leaf-2, and Leaf-3. Figure S4 Phylogenetic tree analysis of polyketide synthase gene families. CHS and STS identified in this study are indicated by red dots, and the previously reported polyketide synthase genes in Polygonum cuspidatum are indicated by blue dots. Figure S5 The types of differentially expressed TFs. Figure S6 Heat map analysis of MYB and WRKY expression in different tissues. Figure S7 HPLC spectrum of standard substances of resveratrol and polydatin. Figure S8 Standard curves of (A) polydatin, (B) resveratrol, and (C) anthraquinones.

Objective: To investigate the effects of Maytenus compound on the proliferation of hepatocellular carcinoma (HCC) cells in vitro and in vivo and to explore the underlying mechanism. Methods: The half maximal inhibitory concentration (IC50) values of Maytenus compound in HepG2 and BEL-7402 cells were determined by the MTS assay. HepG2 and BEL-7402 cells were treated with different concentrations of Maytenus compound. MTS assays, colony formation assays and cell cycle analyses were performed to clarify the inhibitory effect of Maytenus compound on the proliferation of HepG2 and BEL-7402 cells in vitro. After subcutaneous injection of HepG2 cells, nude mice were randomly divided into a vehicle control group and a drug intervention group, which were intragastrically administered ddH2O or Maytenus compound, respectively. The inhibitory effect of Maytenus compound on the proliferation of HepG2 cells in vivo was analyzed using subcutaneous tumor growth curves, tumor weight, the tumor growth inhibition rate and the immunohistochemical detection of BrdU-labeled cells in S phase. The organ toxicity of Maytenus compound was initially evaluated by comparing the weight difference and organ index of the two groups of nude mice. The main proteins in the EGFR-PI3K-AKT signaling pathway were detected by Western blot after Maytenus compound intervention in vivo and in vitro. Results: Maytenus compound showed favorable antiproliferation activity against HepG2 and BEL-7402 cells with IC50 values of 79.42±11.71 µg/mL and 78.48±8.87 µg/mL, respectively. MTS assays, colony formation assays and cell cycle analyses showed that Maytenus compound at different concentration gradients within the IC50 concentration range significantly suppressed the proliferation of HepG2 and BEL-7402 cells in vitro and inhibited cell cycle progression from G1 to S phase. Additionally, Maytenus compound, at an oral dose of 2.45 g/kg, dramatically inhibited, without obvious organ toxicity, the proliferation of subcutaneous tumors formed by HepG2 cells in nude mice. In addition, the tumor growth inhibition rate for Maytenus compound was 66.94%. Furthermore, Maytenus compound inhibited the proliferation of liver orthotopic transplantation tumors in nude mice. Western blot analysis showed that Maytenus compound significantly downregulated the expression of p-EGFR, p-PI3K, and p-AKT and upregulated the expression of p-FOXO3a, p27, and p21 in vivo and in vitro. Conclusion: Maytenus compound significantly inhibited the proliferation of HCC cells in vitro and in vivo. The downregulation of the EGFR-PI3K-AKT signaling pathway and subsequent inhibition of cell cycle progression from G1 to S phase is one of the possible mechanisms. Maytenus compound has a high tumor growth inhibition rate and has no obvious organ toxicity, which may make it a potential anti-HCC drug, but the results from this study need to be confirmed by further clinical trials in HCC patients.

A new alkaloid, tournesibirin (

Melanoma is characterized by high mortality and poor prognosis due to metastasis. AFF4 (AF4/FMR2 family member 4), as a scaffold protein, is a component of the super elongation complex (SEC), and is involved in the progression of tumors, e.g., leukemia, head and neck squamous cell carcinoma (HNSCC). However, few studies on AFF4 have focused on melanoma. Here, AFF4 expression levels and clinicopathological features were evaluated in melanoma tissue samples. Then, we performed cell proliferation, migration and invasion assays in A375 and A2058 cells lines in vitro to evaluate the role of AFF4 in melanoma. The effects of AFF4 knockdown in vivo were characterized via a xenograft mouse model. Finally, the correlation between c-Jun and AFF4 protein levels in melanoma was analyzed by rescue assay and immunohistochemistry (IHC). We found that AFF4 expression was upregulated in melanoma tumor tissues and that AFF4 protein expression was also closely related to the prognosis of patients with cutaneous melanoma. Moreover, AFF4 could promote the invasion and migration of melanoma cells by mediating epithelial to mesenchymal transition (EMT). AFF4 might regulate c-Jun activity to promote the invasion and migration of melanoma cells. Importantly, c-Jun was regulated by the AFF4 promoted melanoma tumorigenesis in vivo. Taken together, AFF4 may be a novel oncogene that promotes melanoma progression through regulation of c-Jun activity.

Herein, we have demonstrated that the plasma-exfoliation of CoOOH nanolayers (∼8 nm) into an ultrathin structure (∼2 nm) could significantly improve the PEC activities of BiVO4 photoanodes. A photocurrent density of 4.9 mA cm−2 and a charge separation efficiency of 89% have been achieved at 1.23 VRHE under AM 1.5G illumination, which are more than two-fold higher than those of pristine CoOOH/BiVO4 photoanodes.