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HYR-PB21 is a new sustained-release formulation of bupivacaine indicated for controlling postoperative pain. The objectives of this study were to investigate the analgesic efficacy and safety profile of HYR-PB21 in patients after haemorrhoidectomy.This was a multicentre, randomised, double-blind, positive-controlled trial. Patients were assigned randomly to receive a single dose of HYR-PB21 (150 mg or 300 mg) or bupivacaine HCl (75 mg) after surgery for prolapsing haemorrhoids. Postoperative pain was evaluated using a numeric rating scale at rest to calculate a cumulative pain score. Total rescue opioid usage and the proportion of subjects receiving rescue opioid were also assessed.We enrolled 72 patients with haemorrhoidectomy, and 71 patients completed the study. The average cumulative pain score through 72 h after surgery in the 300 mg HYR-PB21 group (87 scores) was lower than in the bupivacaine HCl group (166 scores) in an intention-to-treat analysis (P0.001). There was a dose-response effect in reducing total opioid usage and the proportion of rescue opioid use between the 150 mg and 300 mg HYR-PB21 groups, with bupivacaine HCl as a reference group. The HYR-PB21 groups did not show more adverse effects than the bupivacaine HCl group.Local infiltration of a single dose of HYR-PB21 sustained-release bupivacaine had better efficacy in controlling postoperative pain, with similar adverse effects, compared with a single dose of bupivacaine HCl in patients after haemorrhoidectomy.ChiCTR2000041318 (Chinese Clinical Trial Registry).

Cytochrome P450 monooxygenase (P450 or CYP) plays an important role in the metabolism of insecticides and plant allelochemicals by insects. CYP321B1, a novel Spodoptera litura P450 gene, was identified and characterized. CYP321B1 contains a 1488 bp open reading frame (ORF) that encodes a 495 amino acid protein. In fourth instar larvae, the highest CYP321B1 expression levels were found in the midgut and fat body. In the tannin feeding test, tannin can significantly induce the expression of CYP321B1 in the midgut and fat body of 4th instar larvae. To verify the function of CYP321B1, RNA interference and metabolome analysis were performed. The results showed that silencing CYP321B1 significantly reduced the rate of weight gain under tannin induction. Metabolome analysis showed silencing affected 47 different metabolites, mainly involved in secondary metabolite biosynthesis and amino acid metabolism, including amino acids, lipid fatty acids, organic acids and their derivatives. Henoxyacetic acid and cysteamine are the most highly regulated metabolites, respectively. These findings demonstrate that CYP321B1 plays an important role in tannin detoxification and metabolism. Functional knowledge about metabolite detoxification genes in this major herbivorous insect pest can provide new insights into this biological process and provide new targets for agricultural pest control.

Propylea japonica has been regarded as one of the most remarkable natural enemies against aphid in China. However, the mechanism of juvenile hormone (JH) regulation of reproduction in P. japonica is still unclear. In this study, we investigated the JH titers of P. japonica and the development of the ovaries. We selected the six different developmental stages of ladybeetle females for transcriptome sequencing. We identified 583 genes involved in insect reproduction regulation, including 107 insect hormone synthesis signaling pathway-related genes and 476 nutrition-sensing signaling pathway-related genes. Transcriptome analysis indicated that a large number JH synthesis- and metabolism-related enzyme genes and some potential nutrient signal sensing- and transduction-related genes were significantly differentially expressed during P. japonica development. We investigated the effects of Met gene silencing on the reproduction of female adults and found that the ovarian maturation, vitellogenesis, and follicular epithelium development in the dsMet treatment group were significantly inhibited.

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.

In insects, cytochrome P450 monooxygenases (P450s or CYPs) play an important role in the detoxification and metabolism of exogenous plant allelochemicals. In this study, a P450 gene CYP6AB12 was identified and characterized from Spodoptera litura. The cDNA contains an open reading frame (ORF) encoding 511 amino acid residues. CYP6AB12 was expressed at different ages of S. litura, with the highest levels found in the third and fourth instar larvae. Its highest expression was found in the midgut and fat body of fourth instar larvae fed with gossypol. Moreover, these expression levels were substantially increased compared with those from larvae fed with control diet. Gene silencing was then conducted by smearing dsRNA mixed with nanomaterials onto the cuticle. CYP6AB12 expression was significantly decreased in the midgut and fat body, and the net weight increase was substantially lower than that of the control group, indicating that the treatment group had more sensitivity to gossypol than the control. These results reveal that CYP6AB12 plays an important role in the detoxification and metabolism of gossypol, thus further confirming that P450s have a broad ability to detoxify and metabolize plant allelochemicals. It provides an important molecular basis for the exploration of detoxification metabolism and pest control of S. litura.

Elevated CO2 concentrations can change the multi-level nutritional relationship of the ecosystem through the cascading effect of the food chain. To date, few studies have investigated the effects of elevated CO2 concentration on the Araneae species through the tritrophic system. Hylyphantes graminicola (Araneae: Linyphiidae) is distributed widely in Asia and is a dominant predator in cotton fields. This study investigated chemical components in the food chain of cotton (Gossypium hirsutum)—cotton aphid (Aphis gossypii)—predator (H. graminicola) and compared the development, reproduction, and predation of H. graminicola under ambient (400 ppm) and elevated concentration of CO2 (800 ppm). The results showed that the elevated CO2 concentration increased the chemicals of cotton and cotton aphid, but it did not affect the nutrients, development, reproduction, and predation of the spider. However, the survival rate of the spider was significantly decreased in elevated CO2. The results will further our understanding of the role of natural enemies in an environment with elevated CO2 concentration.

Lysiphlebia japonica (Ashmead) is a predominant parasitoid of cotton-melon aphids in the fields of northern China with a proven ability to effectively control cotton aphid populations in early summer. For accurate normalization of gene expression in L. japonica using quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR), reference genes with stable gene expression patterns are essential. However, no appropriate reference genes is L. japonica have been investigated to date. In the present study, 12 selected housekeeping genes from L. japonica were cloned. We evaluated the stability of these genes under various experimental treatments by RT-qPCR using four independent (geNorm, NormFinder, BestKeeper and Delta Ct) and one comparative (RefFinder) algorithm. We identified genes showing the most stable levels of expression: DIMT, 18S rRNA, and RPL13 during different stages; AK, RPL13, and TBP among sexes; EF1A, PPI, and RPL27 in different tissues, and EF1A, RPL13, and PPI in adults fed on different diets. Moreover, the expression profile of a target gene (odorant receptor 1, OR1) studied during the developmental stages confirms the reliability of the chosen selected reference genes. This study provides for the first time a comprehensive list of suitable reference genes for gene expression studies in L. japonica and will benefit subsequent genomics and functional genomics research on this natural enemy.

Risk assessments of insecticidal Cry proteins from Bacillus thuringiensis (Bt) cotton in soils have been mainly based on microcosm and short-term experiments. There is a lack of longer-term field results, especially when Bt-cotton residues are incorporated into the soil. In this study, we used a six-year field trial to evaluate how cultivation duration and Cry1Ac-inputted modes of Bt-cotton affect the persistence of Cry1Ac proteins and soil microbe-mediated enzymatic properties. The results showed that the persistence of Cry1Ac proteins increased with cultivation duration and periodic residue incorporation of the transgenic Bt-cotton variety ZM41. Moreover, temporal residue incorporation had a relatively larger contribution to the persistence of Cry1Ac proteins in the soil than their release in the growth period. Regardless of Bt-cotton cultivation or residue incorporation, soil microbial biomass was significantly suppressed. However, the dehydrogenase activity was significantly stimulated in Bt-cotton cultivation but suppressed in residue incorporation. The activities of β -glucosidase, nitrate reductase, phosphomonoesterase and arylsulfatase were significantly stimulated in soils with Bt-cotton residue incorporation. Based on a structural equation model analysis, the change in enzymatic activity of these four enzymes was attributed to both a direct effect from Cry1Ac proteins and an indirect effect via dehydrogenase.

The extraction of Mg2+ by di-(2-ethylhexyl) phosphoric acid (P204) was carried out to separate lithium and magnesium. To facilitate the direct electrodeposition of metallic magnesium from the magnesium-rich organic phase, different diluents and ionic liquids were investigated. It was found that tributylhexylphosphonium bis (trifluoromethyl sulfonyl) imide ionic liquid ([P4446][NTf2]) can serve as an excellent supporting electrolyte to solve the electroconductibility of the weak polarity organic phase. The electroconductivity was further increased by more than 1500 times by the use of methyl isobutyl ketone (MIBK) as a diluent. The single-stage extraction rate of Mg2+ at a concentration up to 76.87 g/L reached 68.4% using 40% P204 + 55% MIBK + 5% [P4446][NTf2] at R(O/A) = 6:1, and the electroconductivity of organic phase was 254.5 μS·cm−1 after extraction. The extraction mechanism and thermokinetics indicated that both [P4446][NTf2] and MIBK do not participate in the extraction process, and the extraction is an obvious endothermic process.

Common varieties of genetically modified (GM) cotton increasingly display insect-resistant properties via expression of bacterial-derived toxins from Bacillus thuringiensis (Bt). This necessitates a deeper understanding of the possible effects of these crops on non-target insects. The mirid bug Apolygus lucorum is a major pest in cotton production in China, however, the effect of GM cotton on this non-target species is currently virtually unknown. This insect is exposed to these transgenic plants by consuming genetically modified (GM) leaves. In this study, laboratory experiments were conducted to assess the toxicity of CCRI41 and CCRI45, (genetically modified cotton varieties which express the toxins Cry1Ac and CpTI (Cowpea Trypsin Inhibitor)) on nymphs and adults of A. lucorum. There was no detectable increase in mortality after A. lucorum fed on GM cotton leaves for 20 days. While we detected trace amounts of Cry1Ac proteins in both A. lucorum nymphs and adults (10 ng/g fresh weight), the expression of genes related to detoxification did not detectably differ from those feeding on non-GM cotton. Our binding assays did not show Cry1Ac binding to receptors on the midgut brush border membrane from either A. lucorum nymphs or adults. Our findings collectively indicate that feeding on leaves of the GM cotton varieties CCRI41 and CCRI45 have few toxic effects on A. lucorum.

As a new-type liquid lithium resource, the recovery of lithium from geothermal water is of great significance due to the increasing importance of lithium in modern industry. In this work, different lithium manganese oxides (LMOs), including LiMn2O4, Li1.66Mn1.66O4, and Li4Mn5O12, were synthesized and applied for selective recovery of lithium from geothermal water. It was found that the powdery Li4Mn5O12 exhibits high performances for low concentrations of Li+ and was then successfully granulated using chitosan (CTS) at a mass ratio of CTS/LMO = 3:2. The hydrophily of CTS leaded to a serious dissolution loss of CTS/LMO under acidic conditions, but it was successfully solved by direct cross-link of the spherical material using ethylene glycol diglycidyl ether (EGDE), by which the dissolution loss in 0.25 mol/L HCl was decreased from 27.65% to no more than 0.6%. The equilibrium adsorption capacity of Li+ in geothermal water reached 8.98 mg/g at 303.15 K, while the attenuation of adsorption capacity was no more than 1.1% after five times adsorption-desorption cycles. The separation factors between Li+ and other ions were all higher than 51 with a distribution coefficient of Li+ reaching up to 1113.27 mL/g. All these properties demonstrate that the developed material has high selectivity and stability, and can be used as a candidate for selective recovery of lithium from geothermal water or other solutions.

Gene-editing techniques, such as TALEN and CRISPR/Cas9, have been widely used for target DNA editing in many research fields. However, how to rapidly screen and identify the expected gene-edited products with high efficiency and low cost is still a difficult task. Here, we report the development and optimization of one such method that combines quantitative PCR with high-resolution melting analysis (qPCR-HRM) to screen and identify CRISPR/Cas9-edited rice plants. The results showed that gene-edited rice plants with small target DNA in/dels or even single base pair insertion/deletions could be successfully identified. The sensitivity of the qPCR-HRM method is as low as 1%, which is satisfying to be used for quantitative evaluation of gene-editing efficiency. Sanger sequencing results confirmed that the qPCR-HRM method also has high accuracy. It is concluded that the developed qPCR-HRM method is reproducible, accurate, and efficient for quick screening and identification of gene-edited rice plants.

Insulin signalling in insects, as in mammals, regulates various physiological functions, such as reproduction. However, the molecular mechanism by which insulin signals orchestrate ovarian stem cell proliferation, vitellogenesis, and oviposition remains elusive. Here, we investigate the functions of the phosphoinositide 3-kinase (PI3K)-serine/threonine kinase (Akt) pathway, GTPase Ras/mitogen-activated protein kinase (MAPK) pathway, and their downstream messengers in a natural predator, Chrysopa pallens, by the RNAi method. When C. pallens vitellogenin gene 1 (CpVg1) expression was knocked down, the follicle maturation was arrested and total fecundity was reduced. Silencing C. pallens insulin receptor 1 (CpInR1) suppressed Vg transcription and reduced egg mass and hatching rate. Depletion of C. pallens insulin receptor 2 (CpInR2) transcripts lowered Vg transcript level, hampered ovarian development and decreased reproductive output. Knockdown of C. pallens Akt (CpAkt) and C. pallens extracellular-signal-regulated kinase (Cperk) caused phenotypes similar to those caused by knockdown of CpInR2. Disruption of C. pallens transcription factor forkhead box O (CpFoxO) expression caused no significant effects on ovarian development, but sharply impaired total fecundity. Interference with the expression of C. pallens target of rapamycin (CpTor) gene and C. pallens cAMP-response element binding protein (CpCreb) gene led to a down-regulation of Vg transcription, blocking of ovariole growth, and decrease in egg quality. These results suggested the two CpInRs orchestrate oogenesis and oviposition via two signalling pathways to guarantee natural reproduction in the green lacewing, C. pallens.

Molecular characterization of exogenous DNA integrations in host genome is a key aspect in risk assessment of bioengineered crops. However, gaining a clear understanding of the molecular characters of a bioengineered crop using conventional techniques remains a challenging task. Herein, we report the full molecular characterization of one new transgenic rice event G6H1 via a paired-end sequencing approach and bioinformatics analysis pipelines. Also, the molecular characterization reported was validated using conventional PCR, Sanger sequencing, and digital PCR. The results showed there is only one copy of the exogenous DNA inserted, which is located within chromosome 7 of the G6H1 genome. There is no other unintended integration of sequences from the transformation plasmid. These results indicated that the paired-end sequencing approach, combined with bioinformatics pipeline developed, is well suited to elucidate the molecular characteristics of bioengineered crops, and is efficient, low cost, and comprehensive.

Propylea japonica is a very important predator in agricultural ecosystems, which could be exposed to Bt protein. In this study, the bacterial community of P. japonica fed with normal food and food containing Cry2Ab protein was characterized for the first time using qPCR and high-throughput sequencing approaches. Results showed no effect of Cry2Ab on P. japonica development and reproduction. The most abundant bacterial phylum was Firmicutes, and the most abundant genus was Staphylococcus. The total bacteria copy number was not significantly different across four larval stages. Bacteria species composition was gathered more closely in feed on sucrose solution (sucrose-fed) than in larvae only fed on pea aphid (aphid-fed), the diversity indices of some operational taxonomic unit (OTU) were significantly different between sucrose-fed and aphid-fed samples. Different instar larval stages of P. japonica fed with sucrose solution containing Cry2Ab Bt protein and found no effect on microbial community composition and total bacteria copy numbers. However, effects on relative abundance of microbes, copy numbers of Corynebacterium 1 and Glutamicibacter arilaitensis were observed significantly lower in Bt-fed first and fourth larval stages. Low and high concentrations of Cry2Ab protein altered the microbial abundance relative to sucrose-fed P. japonica and copy numbers of G. arilaitensis and Staphylococcus xylosus were significantly lower in Bt-fed samples than control sucrose-fed. Our results are the first report showing that feeding on Cry2Ab protein does not alter microbial species composition in P. japonica, but effects gene copy number of some dominant bacteria. Further investigations are needed to assess the effect of copy number change on P. japonica.