Search

In insects, male accessory gland proteins (ACPs) are important reproductive proteins secreted by male accessory glands (MAGs) of the internal male reproductive system. During mating, ACPs are transferred along with sperms inside female bodies and have a significant impact on the post-mating physiology changes of the females. Under sexual selection pressures, the ACPs exhibit remarkably rapid and divergent evolution and vary from species to species. The diamondback moth, Plutellaxylostella (L.) (Lepidoptera: Plutellidae), is a major insect pest of cruciferous vegetables worldwide. Mating has a profound impact on the females’ behavior and physiology in this species. It is still unclear what the ACPs are in this species. In this study, two different proteomic methods were used to identify ACPs in P. xylostella. The proteins of MAGs were compared immediately before and after mating by using a tandem mass tags (TMT) quantitative proteomic analysis. The proteomes of copulatory bursas (CB) in mated females shortly after mating were also analyzed by the shotgun LC-MS/MS technique. In total, we identified 123 putative secreted ACPs. Comparing P.xylostella with other four insect ACPs, trypsins were the only ACPs detected in all insect species. We also identified some new insect ACPs, including proteins with chitin binding Peritrophin-A domain, PMP-22/ EMP/ MP20/ Claudin tight junction domain-containing protein, netrin-1, type II inositol 1,4,5-trisphosphate 5-phosphatase, two spaetzles, allatostatin-CC, and cuticular protein. This is the first time that ACPs have been identified and analyzed in P. xylostella. Our results have provided an important list of putative secreted ACPs, and have set the stage for further exploration of the functions of these putative proteins in P.xylostella reproduction.

To explore the technique of autogenous bone graft combined with plate fixation in total knee arthroplasty(TKA) with severe proximal medial tibial bone defect.From March 2012 to October 2018, 21 patients (9 males and 12 females) with severe bone defects in the proximal medial tibia during primary total knee arthroplasty were treated with autogenous structural bone grafting and steel plate fixation, with an age of 61 to 77 years old with an average of (69.6±9.1) years and a course of 64 to 257 months with an average of (73.6±170.7) months. According to Rand classification, there were 13 cases of type Ⅲb and 8 cases of type Ⅳb. Postoperative complications were observed, and knee joint function was evaluated by the Hospital for Special Surgery (HSS) score and SF-36 quality of life score.All 21 patients were followed up for 37 to 64 months with an average of (49.5±13.7) months. The incisions of all patients healed smoothly, and 2 patients developed lower limb intermuscular venous plexus thrombosis after operation. There were no periprosthetic infection, loosening of prosthesis and other complications. The autogenous bone grafts of all patients achieved bony healing during postoperative X-ray follow-up, and the healing time was 8 to 13 months with an average of (10.1±2.3) months. The HSS score of patients increased significantly from 30 to 48 with an average of (53.4±4.2) before operation to 75 to 92 with an average of (81.2±8.4) at the final follow-up (The technique of autogenous bone graft combined with steel plate fixation can achieve satisfactory osseointegration effect in the treatment of severe proximal tibial bone defects in primary knee arthroplasty, with less complications and obvious improvement in knee function.

Crop resistance and biological control are both considered efficient and environmentally friendly methods of sustainable pest control. In this study, we aimed at investigating the direct influence of four wheat lines with varying resistance level on the life-history traits of the greenbug, Schizaphis graminum, and the mediational effect on the functional response of a predatory ladybird, Propylaea japonica, under laboratory conditions. Results showed that the aphid fitness was the lowest for aphids that had been feeding on wheat line ‘98-10-19’ for one year. These aphids had the longest development time, and least adult mass, minimal mean relative growth rate, and lowest reproductive fitness. In contrast, the aphids that fed on wheat line ‘98-10-30’ were the fittest, with the shortest development time and highest levels of reproductive fitness. The predatory activities of the ladybeetle, especially the adult male significantly decreased following the consumption of aphids belonging to the ‘98-10-19’-acclimated population. However, there were no significant differences in predatory efficiency (net attack frequency) among the four aphid acclimated populations. Our results showed that the wheat line ‘98-10-19’ has a relative higher resistance to S. graminum than the other three wheat lines, which could further decrease the amount of prey available for consumption. However, the ecological effect of the resistance of ‘98-10-19’ to S. graminum posed no negative influence on the biocontrol potential of P. japonica to these aphids, as their predatory efficiency increases at the fourth instar larvae phase.

Crop resistance and biological control are both considered efficient and environmentally friendly methods of sustainable pest control. In this study, we aimed at investigating the direct influence of four wheat lines with varying resistance level on the life-history traits of the greenbug

To discuss the result and long time follow-up of unstable intertrochanteric fracture combined with avascular necrosis of femoral head treated by total hip arthroplasty.From March 2008 to October 2014, 23 cases were enrolled and treated by total hip arthroplasty, including 10 males and 13 females, aged 59 to 82 years old with an average of(68.4±10.4) years old. There were 8 cases of type Ⅱa fractures, 6 cases of type Ⅱb fractures, 9 cases of type Ⅲ fractures according to Evans Classification. There were 3 cases of type Ⅱa, 3 cases of type Ⅱb, 9 cases of type Ⅲ, 8 cases type Ⅳaccording to Ficat Classification. The complications were recorded, the joint function was evaluated by Harris score and prosthesis survival rate was analysed.Postoperative complications occurred in 2 cases, one patient had acute periprosthetic infection 3 months after operation, and one patient had hip dislocation 3 months after operation. All 23 patients were followed up for 61 to 110 months with an agerage of (85.1±22.9) months. The Harris score at the final follow-up was 83 to 92 months with an average of(89.8±5.2) months, which was significantly different from that before operation (The symptoms of unstable intertrochanteric fracture combined with avascular necrosis of femoral head could be solved by total hip arthroplasty, and the long time result was satisfied.

To explore clinical effects of carpal canal endoscopy in treating patients with plantar fasciopathy who failed by conservative treatment.From August 2018 to August 2019, 50 patients with plantar fascia were divided into two groups and 25 patients in each group. In carpal canal endoscopy group, included 11 males and 14 females, aged from 39 to 67 years old with an average of(57.7±6.4) years old;carpal canal endoscopy was used to plantar fascia release. In arthroscopy group, included 9 males and 16 females, aged from 41 to 73 years old with an average of (58.1±7.2) years old;conventional 4.0 mm arthroscopy Instruments was used to plantar fascia release. Operation time, hospitalization expense and postoperative complications between two groups were observed and compared. Postoperative visual analogue scale(VAS) and American Orthopedic Foot Ankle Society (AOFAS) score were used to evaluate clinical function.All patients were followed up from 12 to 18 months with an average of (14.3±2.1) months. There were significant differentces in operation time and hospitalization expense between two groups (The outcome of carpal canal endoscopy and arthroscopy has similar effects in treating plantar fascia. While carpal canal endoscopy has advantages of need not perfusion during opertaion, protect soft tissue well, less operation time, and lower cost.

Plant resistance against insects mainly depends on nutrient restriction and toxic metabolites, but the relative importance of nutrition and toxins remains elusive. We examined performance, nutrition ingestion, and transcriptome response of the English grain aphid, Sitobion avenae, feeding on resistant Xiaoyan 22 (XY22) and susceptible Xinong 979 (XN979) wheat cultivars. Aphids had lower body weight and fecundity when feeding on XY22 than on XN979, although the phloem sap of XY22 had a higher nutritive quality (in terms of amino acid:sucrose ratio). Aphids feeding on XY22 also had a lower honeydew excretion rate than those on XN979, suggesting that aphids ingested less phloem sap from XY22. The transcriptome data showed 600 differentially expressed genes (DEGs), and 11 of the top 20 KEGG pathways significantly enriched in DEGs were involved in nutrient metabolism. We found 81 DEGs associated with the metabolism of sugars, lipids, and amino acids, 59 of which were significantly downregulated in aphids feeding on XY22. In contrast, there were 18 DEGs related to detoxifying metabolism, namely eight UDP-glucuronosyltransferases, six cytochromes P450 monooxygenases, one glutathione S-transferase, two ATP-binding cassette transporters, and one major facilitator superfamily transporter; 12 of these were upregulated in the aphids feeding on XY22. Our results indicated that both the quantity and quality of phloem nutrition available to aphids are critical for the growth and development of aphids, and the higher resistance of XY22 is mainly due to the reduction in phloem sap ingested by aphids, rather than toxic metabolites.

To investigate the clinical effect of using lengthened trochanteric osteotomy wire fixation combined with autologous bone graft in patients undergoing revision total hip arthroplasty.From December 2010 to December 2018, 18 patients underwent revision of total hip arthroplasty with extended trochanteric osteotomy wire fixation and autogenous bone graft, including 8 males and 10 females with an average age of (78.89±3.32) years old ranging from 68 to 82 years. The time from the initial replacement to the revision was 9 to 22 (16.33±2.93) years. The patients were followed up regularly after operation. The healing time of osteotomy, the time of full weight-bearing activity, Harris score of hip joint and complications were recorded.All 18 patients were followed up for 16 to 38 months with an average of (25.78±6.65) months. The incision length was 16 to 21 cm with an average of (18.89±1.32) cm; the operation time was 105 to 128 min with an average of (115.44±6.59) min, the bleeding volume was 240 to 285 ml with an average of (267.44±13.77) ml. The healing time of osteotomy was 12 to 18 weeks with an average of (15.61±1.75) weeks. Harris score of hip joint was (47.11±5.04) before operation, (76.39±3.85) during full weight-bearing activities, and (82.22±2.76) at the final follow-up(In revision total hip arthroplasty, the use of extended trochanteric osteotomy wire fixation combined with autologous bone graft can achieve satisfactory clinical results, but the surgeon needs to make a systematic plan for the pre-revision, intraoperative and postoperative recovery.

To investigate the value of lumbar quantitative CT (QCT) in vertebroplasty for osteoporotic fracture combined with scoliosis.The clinical data of 60 patients with osteoporotic fractures combined with different degrees of scoliosis treated by vertebroplasty from December 2017 to December 2019 were retrospectively analyzed. There were 18 males and 42 females, aged from 65 to 81 (72.63±3.34)years old. All patients were received QCT examination before surgery. According to the QCT value, the patients were divided into osteopenia group(QCT80 g/L, 10 cases, 12 vertebrae), osteoporosis group(QCT 40-80 g/L, 35 cases, 48 vertebrae) and severe osteoporosis group(QCT40 g/L, 15 cases, 22 vertebrae). The dispersion and leakage of bone cement in the injured vertebrae of patients with different degrees of QCT value were observed, and the QCT value in the selection of puncture point, correction of Cobb angle and recovery of vertebral height were analyzed in the patients.Among 60 cases of 82 vertebrae, 41 cases of 55 vertebrae were punctured by concave unilateral puncture, according for 67.07%. Among them, there were 2 cases with 2 vertebrae in osteopenia group, 26 cases with 35 vertebrae in osteoporosis group, and 13 cases with 18 vertebrae in severe osteoporosis group. There was significant difference in the number of cases with unilateral or bilateral puncture among the three groups (For patients with osteoporotic fracture combined with scoliosis undergoing vertebroplasty, the severity of osteoporosis should be determined according to lumbar QCT detection, and the concave side of scoliosis should be selected for puncture, which is conducive to improving scoliosis, restoring spinal stability and improving surgical safety.

NADPH-cytochrome P450 reductase (CPR) is involved in the metabolism of endogenous and exogenous substances, and detoxification of insecticides. RNA interference (RNAi) of CPR in certain insects causes developmental defects and enhanced susceptibility to insecticides. However, the CPR of Acyrthosiphon pisum has not been characterized, and its function is still not understood. In this study, we investigated the biochemical functions of A. pisum CPR (ApCPR). ApCPR was found to be transcribed in all developmental stages and was abundant in the embryo stage, and in the gut, head, and abdominal cuticle. After optimizing the dose and silencing duration of RNAi for downregulating ApCPR, we found that ApCPR suppression resulted in a significant decrease in the production of cuticular and internal hydrocarbon contents, and of cuticular waxy coatings. Deficiency in cuticular hydrocarbons decreased the survival rate of A. pisum under desiccation stress, and increased its susceptibility to contact insecticides. Moreover, desiccation stress induced a significant increase in ApCPR mRNA levels. We further confirmed that ApCPR participates in cuticular hydrocarbon production. These results indicate that ApCPR modulates cuticular hydrocarbon production, desiccation tolerance, and insecticide susceptibility in A. pisum, and presents a novel target for pest control. This article is protected by copyright. All rights reserved.

Crop resistance plays a role in preventing aphid damage, benefiting food production industries, but its effects are limited due to aphid adaptation and phenotypic plasticity. Therefore, furthering understanding of aphid–crop interactions will improve our ability to protect crops from aphids. To determine how aphids adapt to resistant varieties of wheat, Triticum aestivum L. over time, we performed a laboratory experiment to assess the multi-generational effects of three wheat varieties, Batis, Ww2730, and Xiaoyan22, with different resistance levels on the fitness of Sitobion avenae (Fab.) (Hemiptera: Aphididae). The results showed that Ww2730 and Xiaoyan22 were more resistant than Batis to S. avenae, regardless of whether the aphids were newly introduced or had been acclimated before being introduced to the three wheat varieties. However, the effect of resistance on aphid life-history traits was time dependent. Aphid weigh gain increased and they development faster of the acclimated generation compared to the newly introduced generation on all three varieties. And the fecundity on the three varieties and net reproduction rates on Batis and Xiaoyan22 significantly decreased. Aphid fitness in terms of individual life-history parameters improved, whereas aphid fitness in terms of reproductive decreased, and a convergence effect, the difference gaps and standard errors of all life-history traits among the three acclimated populations had narrowed and were less than those in the three first-generation populations, was observed during the 3-mo experimental period. We suggested that S. avenae could rapidly respond to wheat resistance through life-history plasticity.

To investigate the clinical effect of double plate combined with iliac bone graft in the treatment of femoral nonunion after intramedullary nailing.From December 2008 to December 2017, double plate combined with autogenous iliac bone graft was used to treat femoral nonunion after intramedullary nailing. There were 11 cases, including 10 males and 1 female, aged 35 to 62 years, and the time from fracture to nonunion was 12 to 20 months. According to Judet classification, there were 8 cases of atrophic nonunion and 3 cases of proliferative nonunion. Regular follow-up was conducted after operation to record the fracture healing time, load-bearing activity time and complications, and to observe the repair effect of double plate fixation combined with iliac bone graft on nonunion after femoral shaft fracture operation.All patients were followed up for 12 to 22 months. The operation time was 70 to 130 min and the blood loss was 180 to 350 ml. After operation, 2 cases had knee stiffness, which recovered after passive exercise with CPM machine for 2 weeks;1 case had pain in iliac bone donor area, which was relieved after 3 months. The time of fracture healing was 24 to 40 weeks, and the time of complete weight-bearing activity was 14 to 32 weeks. SF-36 quality of life score at the final follow-up:body pain 70 to 82, activty 70 to 82, social function 72 to 83, the overall health 72 to 82. At the end of the follow-up, there were no complications such as limb shortening, infection, poor wound healing, internal fixation failure (fracture, loosening).It is an effective method to treat nonunion of femur after intramedullary nailing by using double plate combined with autogenous iliac bone graft.

Insect cuticular hydrocarbons (CHCs) serve as important intersexual signaling chemicals and generally show variation between the sexes, but little is known about the generation of sexually dimorphic hydrocarbons (SDHCs) in insects. In this study, we report the molecular mechanism and biological significance that underlie the generation of SDHC in the German cockroach Blattella germanica. Sexually mature females possess more C29 CHCs, especially the contact sex pheromone precursor 3,11-DimeC29. RNA interference (RNAi) screen against the fatty acid elongase family members combined with heterologous expression of the genes in yeast revealed that both BgElo12 and BgElo24 were involved in hydrocarbon (HC) production, but BgElo24 is of wide catalytic activities and is able to provide substrates for BgElo12, and only the female-enriched BgElo12 is responsible for sustaining female-specific HC profile. Repressing BgElo12 masculinized the female CHC profile, decreased contact sex pheromone level, and consequently reduced the sexual attractiveness of female cockroaches. Moreover, the asymmetric expression of BgElo12 between the sexes is modulated by sex differentiation cascade. Specifically, male-specific BgDsx represses the transcription of BgElo12 in males, while BgTra is able to remove this effect in females. Our study reveals a novel molecular mechanism responsible for the formation of SDHCs and also provide evidences on shaping of the SDHCs by sexual selection, as females use them to generate high levels of contact sex pheromone., Sexual dimorphism of body waxes is prevalent in insects; this study reveals that the sex-differentiation pathway regulates fatty acid elongation, ensuring production of the sexually dimorphic cuticular hydrocarbons needed for high levels of sex pheromone and sexual attractiveness in female cockroaches.

Insect cuticular hydrocarbons (CHCs) serve as important intersexual signaling chemicals and generally show variation between the sexes, but little is known about the generation of sexually dimorphic hydrocarbons (SDHCs) in insects. Here, we report the molecular mechanism and biological significance that underling the generation of SDHC in the German cockroach,Blattella germanica. Sexually mature females possess more C29 cuticular hydrocarbons (CHCs), especially the contact sex pheromone precursor 3,11-DimeC29. RNAi screen against fatty acid elongase gene family members and combined with heterologous expression revealed that bothBgElo12andBgElo24were involved in HC production, butBgElo24is of wide catalytic activities and is able to provides substrates forBgElo12, and only the female-enrichedBgElo12was responsible for sustaining female-specific HC profile. RepressingBgElo12masculinized the female CHC profile, decreased contact sex pheromone level and consequently reduced the sexual attractiveness of female cockroaches. Moreover, the asymmetric expression ofBgElo12between the sexes is modulated by sex differentiation cascade. Specifically, male-specificBgDsxrepresses the transcription ofBgElo12in males, whileBgTrais able to remove this effect in females. Our study reveals a novel molecular mechanism responsible for the formation of SDHCs, and also provide evidences on shaping of the SDHCs by sexual selection, as females use them to generate high levels of contact sex pheromone.

Water retention is critical for physiological homeostasis and survival in terrestrial insects. While deposition of hydrocarbons on insect cuticles as a key measure for water conservation has been extensively investigated, we know little about other mechanisms for preventing water loss in insects. Here, we report two fatty acid synthetic genes that are independent of hydrocarbon production but crucial for water retention in the German cockroach Blattella germanica (L.). First, an integument enriched fatty acid elongase gene (BgElo1) was identified as a critical gene for desiccation resistance in B. germanica; however, knockdown of BgElo1 surprisingly failed to cause a decline in cuticular lipids. In addition, RNA interference (RNAi)-knockdown of an upstream fatty acid synthase gene (BgFas3) showed a similar phenotype, and transmission electron microscopy analysis revealed that BgFas3- or BgElo1-RNAi did not affect cuticle architecture. Bodyweight loss test showed that repression of BgFas3 and BgElo1 significantly increased the weight loss rate, but the difference disappeared when the respiration was closed by freeze killing the cockroaches. A water immersion test was performed, and we found that BgFas3- and BgElo1-RNAi made it difficult for cockroaches to recover from drowning, which was supported by the upregulation of hypoxia-related genes after a 10-h recovery from drowning. Moreover, a dyeing assay with water-soluble Eosin Y showed that this was caused by the entry of water into the respiratory system. Our research suggests that BgFas3 and BgElo1 are required for both inward and outward waterproofing of the respiratory system. This study benefits the understanding of water retention mechanisms in insects.

Free amino acids in the phloem sap are the dominant nitrogen source for aphids, but their availability is usually poor. Although some studies have explored the effect of dietary amino acid restriction on aphid performance, little is known about the molecular basis of these effects. Here, we examined the performance and transcriptome of the green peach aphid, Myzus persicae, fed a standard diet (Control diet) or a diet containing 50% of the total amino acids of the Control diet (Half diet). Aphid weight and fecundity were significantly reduced in the Half diet group. Transcriptomic analysis showed that a total of 1460 genes were differentially expressed between the groups were fed on the two diets, which many of them were associated with nutrient and energy metabolism. When feeding on the Half diet, aphids upregulated genes associated with the amino acid biosynthetic pathway (predominantly amino acid biosynthesis genes and some amino acid transporter genes) as well as the cysteine and serine protease genes. Furthermore, these aphids displayed increased expression of genes associated with glycolysis, which could generate intermediates for de novo amino acid biosynthesis. Consistent with this, elevated glucose levels were observed in aphids in the Half diet group. Additionally, the expression levels of several genes associated with hormonal signaling pathway were altered. Several genes related to juvenile hormone and insulin-like peptide (ILP) signaling were downregulated, including Krüppel homolog 1 (Kr-h1) and insulin-like peptide 5 (Ilp5), respectively. In contrast, several genes related to ecdysone signaling were upregulated including broad-complex core protein (Br-c) and shade (Shd). Despite their poor performances, M. persicae adapted to dietary restriction of amino acids, through upregulation of genes involved in amino acid biosynthesis, glycolysis, and protein degradation, as well as by altering the expression level of genes involved in hormone signaling pathways.

Maternal phenotypic regulations between different generations of aphid species help aphids to adapt to environmental challenges. The pea aphid Acyrthosiphon pisum has been used as a biological model for studies on phenotypic regulation for adaptation, and its alternative phenotypes are typically and physiologically based on maternal effects. We have observed an artificially induced and host-related maternal effect that may be a new aspect to consider in maternal regulation studies using A. pisum. Marked phenotypic changes in the cuticular melanization of daughter A. pisum were detected via tyrosine hydroxylase knockdown in the mothers during their period of host plants alternations. This phenotypic change was found to be both remarkable and repeatable. We performed several studies to understand its regulation and concluded that it may be controlled via the dopamine pathway. The downregulation and phenotypes observed were verified and described in detail. Additionally, based on histological and immunofluorescence analyses, the phenotypic changes caused by cuticular dysplasia were physiologically detected. Furthermore, we found that this abnormal development could not be reversed after birth. Transcriptome sequencing confirmed that this abnormal development represents a systemic developmental failure with numerous transcriptional changes, and chemical interventions suggested that transgenerational signals were not transferred through the nervous system. Our data show that transgenerational regulation (maternal effect) was responsible for the melanization failure. The developmental signals were received by the embryos from the mother aphids and were retained after birth. APTH RNAi disrupted the phenotypic determination process. We demonstrate that non-neuronal dopamine regulation plays a crucial role in the transgenerational phenotypic regulation of A. pisum. These results enhance our understanding of phenotyping via maternal regulation in aphids.

Background Cuticular hydrocarbons (CHCs) have a critical role in preventing desiccation and penetration of xenobiotics in insects. Previous studies have shown that cytochrome P450 subfamily 4G (CYP4G) enzymes are oxidative decarbonylases, essential for CHC biosynthesis. However, it is unclear whether there are functional differences between the two CYP4G genes in most insects. In Locusta migratoria, we identified two CYP4G genes (LmCYP4G62 and LmCYP4G102). LmCYP4G102 plays a critical role in the synthesis of CHCs, but the function of LmCYP4G62 is unknown. Results We identified, characterized, and compared two LmCYP4G genes, based on L. migratoria transcriptomic and genomic databases. RT-qPCR showed that both were highly expressed in tissues with which oenocytes are associated, the integument and fat body. Immunostaining indicated that LmCYP4G62 and LmCYP4G102 were highly abundant in oenocytes in these tissues. However, the two enzymes had a different subcellular distribution, with LmCYP4G62 localized on the plasma membrane and LmCYP4G102 dispersed throughout the oenocyte cytoplasm, presumably on the endoplasmic reticulum. RNA interference-mediated gene silencing against each of the two genes resulted in reduced CHC contents, in all classes for LmCYP4G102, but mostly shorter chain CHCs for LmCYP4G62. Silencing of both genes resulted in increased insecticide penetration through the cuticle, and increased locust susceptibility to desiccation and insecticides. Conclusion Our studies suggest that both LmCYP4G62 and LmCYP4G102 contribute to hydrocarbon biosynthesis and play key roles in protecting locusts from water loss and insecticide penetration, but they are not fully redundant. Further, the two LmCYP4G genes might be used as new targets for insect pest management. © 2020 Society of Chemical Industry.

Maternal phenotypic regulations between different generations of aphid species help aphids to adapt to environmental challenges. The pea aphid Acyrthosiphon pisum has been used as a biological model for studies on phenotypic regulation for adaptation, and its alternative phenotypes are typically and physiologically based on maternal effects. We have observed an artificially induced and host-related maternal effect that may be a new aspect to consider in maternal regulation studies using A. pisum. Marked phenotypic changes in the cuticular melanization of daughter A. pisum were detected via tyrosine hydroxylase knockdown in the mothers during their period of host plants alternations. This phenotypic change was found to be both remarkable and repeatable. We performed several studies to understand its regulation and concluded that it may be controlled via the dopamine pathway. The downregulation and phenotypes observed were verified and described in detail. Additionally, based on histological and immunofluorescence analyses, the phenotypic changes caused by cuticular dysplasia were physiologically detected. Furthermore, we found that this abnormal development could not be reversed after birth. Transcriptome sequencing confirmed that this abnormal development represents a systemic developmental failure with numerous transcriptional changes, and chemical interventions suggested that transgenerational signals were not transferred through the nervous system. Our data show that transgenerational regulation (maternal effect) was responsible for the melanization failure. The developmental signals were received by the embryos from the mother aphids and were retained after birth. APTH RNAi disrupted the phenotypic determination process. We demonstrate that non-neuronal dopamine regulation plays a crucial role in the transgenerational phenotypic regulation of A. pisum. These results enhance our understanding of phenotyping via maternal regulation in aphids.

Free amino acids in the phloem sap are the dominant nitrogen source for aphids, but their availability is usually poor. Although some studies have explored the effect of dietary amino acid restriction on aphid performance, little is known about the molecular basis of these effects. Here, we examined the performance and transcriptome of the green peach aphid, Myzus persicae, fed a standard diet (Control diet) or a diet containing 50% of the total amino acids of the Control diet (Half diet). Aphid weight and fecundity were significantly reduced in the Half diet group. Transcriptomic analysis showed that a total of 1460 genes were differentially expressed between the groups were fed on the two diets, which many of them were associated with nutrient and energy metabolism. When feeding on the Half diet, aphids upregulated genes associated with the amino acid biosynthetic pathway (predominantly amino acid biosynthesis genes and some amino acid transporter genes) as well as the cysteine and serine protease genes. Furthermore, these aphids displayed increased expression of genes associated with glycolysis, which could generate intermediates for de novo amino acid biosynthesis. Consistent with this, elevated glucose levels were observed in aphids in the Half diet group. Additionally, the expression levels of several genes associated with hormonal signaling pathway were altered. Several genes related to juvenile hormone and insulin-like peptide (ILP) signaling were downregulated, including Krüppel homolog 1 (Kr-h1) and insulin-like peptide 5 (Ilp5), respectively. In contrast, several genes related to ecdysone signaling were upregulated including broad-complex core protein (Br-c) and shade (Shd). Despite their poor performances, M. persicae adapted to dietary restriction of amino acids, through upregulation of genes involved in amino acid biosynthesis, glycolysis, and protein degradation, as well as by altering the expression level of genes involved in hormone signaling pathways.

Armyworm feeding in large, destructive groups is hugely difficult to control and the oriental armyworm, Mythimna separata (Walk), is one such pest. In this study, we reported a semisynthetic artificial diet for the oriental armyworm. This diet is based on Ritter’s diet, a formula developed for Heliothis zea. The survival of M. separata was extremely low and only around 2% insects can reach the adult stage on Ritter’s diet. But, it can reach up to 100% if corn leaf powder (CLP) was mixed, and insects grew faster and gained more mass. After testing a set of mixtures of Ritter’s diet and CLP, we found that 14.3% was the optimal proportion of CLP for making the artificial diet. We then used chloroform to extract CLP. Insect performance was still much better on Ch-extracted CLP diets than that on Ritter’s diet, but it was poorer than that on the diets containing unprocessed CLP, suggesting that the essential factor(s) was only partially extracted from corn leaf. We then used methanol and dichloromethane, two solvents differing in their polarity, to process the extractions and analyzed the extracted chemicals using gas chromatography–mass spectrometry (GC-MS). Insects had a better performance on dichloromethane-extracted CLP diet in comparison to methanol-extracted one, indicating that the important factor(s) is more prone to methanol extraction. The reported recipe here is useful for the research on M. separata and possibly other grain-crop eating armyworms. The functions of the chemicals extracted from corn leaf tissue can be investigated in the future studies.

Background Insects cannot synthesize sterols and must acquire them from food. The mechanisms underlying how insects uptake dietary sterols are largely unknown except that NPC1b, an integral membrane protein, has been shown to be responsible for dietary cholesterol uptake in Drosophila melanogaster. However, whether NPC1b orthologs in other insect species, particularly the economically important pests, function similarly remains to be determined. Results In this study, we characterized the function of NPC1b in Helicoverpa armigera, a global pest that causes severe yield losses to many important crops. Limiting dietary cholesterol uptake to insects significantly inhibited food ingestion and weight gain. Compared to the wild-type H. armigera, the CRISPR/Cas9-edited NPC1b mutant larvae were incapable of getting adequate cholesterol and died in their early life stage. Gene expression profile and in situ hybridization analyses indicated that NPC1b was mainly expressed in the midgut where dietary cholesterol was absorbed. Expression of NPC1b was also correlated with the feeding life stages and was especially upregulated during early larval instars. Protein-ligand docking and sequence similarity analyses further demonstrated that NPC1b proteins of lepidopteran insects shared a relatively conserved cholesterol binding region, NPC1b_NTD, which, however, was highly divergent from bees-derived sequences. Conclusion NPC1b was crucial for dietary cholesterol uptake and growth of H. armigera, and therefore could serve as an insecticide target for the development of a novel pest-management approach to control this economically significant insect pest with little off-target effect on bees and sterol-autotrophic animals. © 2020 Society of Chemical Industry.

Plants emit volatile organic compounds (VOCs) in response to herbivore attack. VOCs emitted from the Chinese cabbage cultivars in response to the damage by the green peach aphid, Myzus persicae, were unknown. Using a solid-phase microextraction-based headspace collection method, we investigated and compared the emissions of VOCs from seven Chinese cabbage cultivars (Qibao, Qingan 80, Lvlong, Yuanbao, Qingan 70, Jinlv, and Lvqiu 66) in response to M. persicae attack. Our results showed that the VOCs emitted from the cultivars Qingan 80 and Yuanbao differed significantly from the other cultivars in response to the attraction of wingless M. persicae. Most importantly, out of the 27 detected compounds, α-caryophyllene was detected only in Qingan 80 and Qibao, but not in the other five cultivars. Among the compounds detected, 2 monoterpene and 12 terpenes were predominant in all cabbage cultivars. Furthermore, the wingless M. persicae showed preference to Qingan 80 while it had the highest nitrogen content among the tested cultivars. Moreover, we found a remarkable relationship among M. persicae attraction, plant nitrogen content, and total volatile emissions. Nitrogen content of the plants has a significant impact on volatile emission and preference behavior of M. persicae. Our results indicate that the wingless M. persicae were efficient in their interspecific host selection with an ability to distinguish plant cultivar differences by leaf nitrogen content. This study will be helpful in understanding aphid host selection, and sets a stage to further study the attractant-based integrated aphid management programs.

Rising atmospheric carbon dioxide concentration ([CO 2 ]) and warming are two of the most conspicuous characteristics of global climate change in this century, and they are responsible for a variety of physiological responses of plants. However, studies addressing the combined impact of [CO 2 ] and temperature on plant physiological responses are still scarce. We studied the combined effects of long term elevated [CO 2 ] and temperature on dynamic changes of physiological metabolism in cotton with growth and development. Dry matter content (DMC), N concentration, C/N ratio, soluble sugar content, starch content, total nonstructural carbohydrate (TNC), total phenolic concentration and amino acid levels in cotton leaves in response to long term [CO 2 ] and temperature levels were assayed during different growth times. Elevated [CO 2 ] and temperature independently increased DMC at different growth times but the highest increase of DMC occurred under both elevated [CO 2 ] and elevated temperature. Elevated [CO 2 ] reduced leaf N concentration and increased C/N ratio; however, no differences were found at ambient or elevated temperature after both 75 and 105 days of treatments. The leaves grown in elevated [CO 2 ] had higher soluble sugar, starch, TNC contents and total phenolic concentration, whereas lower levels of these components were observed at a higher temperature. There were different interactive effects between [CO 2 ] and temperature on these physiological characteristics at different growth times. Elevated [CO 2 ] increased amino acids of Asp, His, Ile and Thr levels and reduced Glu, Ala and Pro levels at different growth times under ambient temperature. Under elevated temperature, [CO 2 ] enrichment increased Glu and Thr levels, whereas it reduced Ala, Asp and Val levels. There was a significant interactive effect between [CO 2 ] and temperature on 8, 9 and 11 out of the 15 amino acids after 45, 75 and 105 days of treatment, respectively. The results indicate these physiological characteristics in the cotton leaves during different growth times will change differently in response to individual environmental factor or the combination of different environmental factors, and the physiological metabolism in cotton under long term elevated [CO 2 ] and temperature will dynamically change with growth and development, thus they may have different metabolic mechanisms to deal with future climate change. Further work is required to elucidate the mechanisms underlying an interactive effect of [CO 2 ] and temperature on plant physiological and metabolic change as well as the effect of plant nutritional quality changes on plant-herbivore interaction.

Melanism in insects is important for their physical protection, immunoreactions, and sclerotization. The vetch aphid, Megoura viciae (Buckton), has relatively strong tanning in its prothorax, head, antennae, cornicles, and legs. It was hypothesized that M. viciae may sequester the high level of l -DOPA in its host Vicia faba to help in its melanization process for ecological adaptation. To confirm this hypothesis, the amount of l -DOPA in M. viciae was modified and quantified. We first generated a Trifolium repens (clover, low l -DOPA containing) host to cut off the extra l -DOPA intake by M. viciae. The rate-limiting tyrosine hydroxylase gene of M. viciae (MV-TH) was then cloned and analyzed. To further reduce the l -DOPA level in the insect, RNAi was used to downregulate the transcriptional level of MV-TH. Our results confirmed that M. viciae could indeed sequester l -DOPA in its body, and its ample storage of this amino acid could be the reason for the strong tanning of its body. M. viciae reared on T. repens could upregulate its MV-TH to enhance l -DOPA biosynthesis and thus maintain a high level of l -DOPA. The MV-TH repression by RNAi lasted for about 3 days, successfully decreasing the l -DOPA level. Aside from a slight decrease in exuvia tanning, no other obvious change in body appearance was detected in the RNAi-treated insect. Although M. viciae can obtain most of its l -DOPA directly from its original host, its internal l -DOPA synthetase is still functional, especially when extra l -DOPA is removed from the diet. This capability to enhance its shield ensures the ecological adaptation of this insect.

To study clinical effects of double plating through different approaches for communicated and obvious osteoporosis periprosthetic femoral fracture following total knee arthroplasty(TKA).From July 2010 to June 2017, 21 patients with periprosthetic femoral fracture following TKA were divided into two groups according to operative approach. Fifteen patients in medial and lateral double approaches group, including 5 males and 10 females aged from 63 to 79 years old with an average of (67.2±5.9) years old; 11 patients were type 33-A2 and 4 patients were type 33-A3 according to AO-OTA classification; 12 patients injured by falling down and 3 patients by traffic accident; treated with double plating. Six patients in medial parapatellar approach group, including 3 males and 3 females, aged from 61 to 74 years old with an average of (64.6±6.0) years old; 3 patients were type 33-A2 and 3 patients were type 33-A3 according to AO-OTA classification; 5 patients injured by falling down and 1 patient by traffic accident; treated with double plating. Operative time, blood loss, postoperative drainage, fracture healing time were compared between two groups; HSS score and radiology at 3 and 12 months were compared between two groups.All patients were followed up, and the follow-up time of bilateral approaches group ranged from 12 to 18 months with an average of (14.2±2.6 ) months, while the follow-up time of single approach group ranged from 12 to 16 months with an average of (12.6±2.5) months, and there was no statistical difference between two groups. The operative time and postoperative drainage in bilateral approaches group were (107.2±10.4) min and (213.9±30.4) ml, while in sigle approach group was (95.4±12.8) min and (256.8±34.2) ml, and the differences were significant(Double plating technique for communicated and obvious osteoporosis periprosthetic femoral fracture following TKA could obtain good function of knee joint. The medial parapatellar approach has shorter operative time, while the bilateral approaches had less drainage.

In natural ecological systems, plants are often simultaneously attacked by both insects and pathogens, which can affect each other's performance and the interactions can be extended to higher trophic levels, such as parasitoids. The English grain aphid (Sitobion avenae) and powdery mildew (Blumeria graminis f. sp. tritici) are two common antagonists that pose a serious threat to wheat production. Numerous studies have investigated the effect of a single factor (insect or pathogen) on wheat production. However, investigation on the interactions among insect pests, pathogens, and parasitoids within the wheat crop system are rare. Furthermore, the influence of the fungicide, propiconazole, has been found to imitate the natural ecosystem. Therefore, this study investigated the effects of B. graminis on the biological performance of grain aphids and the orientation behavior of its endoparasitic wasp Aphidius gifuensis in the wheat system. Our findings indicated that B. graminis infection suppressed the feeding behavior, adult and nymph weight, and fecundity and prolonged the developmental time of S. avenae. We found that wheat host plants had decreased proportions of essential amino acids and higher content of sucrose following aggravated B. graminis infection. The contents of Pro and Gln increased in the wheat plant tissues after B. graminis infection. In addition, B. graminis infection elicited immune responses in wheat: increase in the expression of defense genes, content of total phenolic compounds, and activity of three related antioxidant enzymes. Moreover, co-infection of B. graminis and S. avenae increased the attraction to A. gifuensis compare to that after infestation with aphids alone. In conclusion, our results indicated that B. graminis infection adversely affected the performance of S. avenae in wheat through restricted nutrition and induced defense response. Furthermore, the preference of parasitoids in such an interactive environment might provide an important basis for pest management control.

In nature, plants have evolved sophisticated defense mechanisms against the attack of pathogens and insect herbivores. Plant volatile-mediated plant-to-plant communication has been assessed in multitrophic systems in different plant species and different pest species. β-ocimene is recognized as an herbivore-induced plant volatile that play an important role in the chemical communication between plants and pests. However, it is still unclear whether β-ocimene can active the defense mechanism of Chinese cabbage Brassica pekinensis against the peach aphid Myzus persicae. In this study, we found that treatment of Chinese cabbage with β-ocimene inhibited the growth of M. persicae in terms of weight gain and reproduction. Moreover, β-ocimene treatment negatively influenced the feeding behavior of M. persicae by shortening the total feeding period and phloem ingestion and increasing the frequency of stylet puncture. When given a choice, winged aphids preferred to settle on healthy Chinese cabbage compared with β-ocimene-treated plants. In addition, performance of the parasitoid Aphidius gifuensis in terms of Y-tube olfaction and landings was better on β-ocimene-treated Chinese cabbage than on healthy plants. Furthermore, β-ocimene significantly increased the expression levels of salicylic acid and jasmonic acid marker genes and the accumulation of glucosinolates. Surprisingly, the transcriptional levels of detoxifying enzymes (CYP6CY3, CYP4, and GST) in aphids reared on β-ocimene-treated Chinese cabbage were significantly higher than those of aphids reared on healthy plants. In summary, our results indicated that β-ocimene can activate the defense response of Chinese cabbage against M. persicae, and that M. persicae can also adjust its detoxifying enzymes machinery to counter the host plant defense reaction.

The maternal effects of the English grain aphid, Sitobion avenae on offspring phenotypes and performance on wheat varieties with different resistance traits were examined. We found that both conditioning wheat varieties(the host plant for over 3 months) and transition wheat varieties affected the biological parameters of aphid offspring after they were transferred between wheat varieties with different resistance traits. The conditioning varieties affected weight gain, development time (DT), and the intrinsic rate of natural increase (rm), whereas transition varieties affected the fecundity, rm, net reproductive rate, and fitness index. The conditioning and transition wheat varieties had significant interaction effects on the aphid offspring’s DT, mean relative growth rate, and fecundity. Our results showed that there was obvious maternal effects on offspring when S. avenae transferred bwteen wheat varieties with different resistance level, and the resistance traits of wheat varieties could induce an interaction between the conditioning and transition wheat varieties to influence the growth, development, reproduction, and even population dynamics of S. avenae. The conditioning varieties affected life-history traits related to individual growth and development to a greater extent, whereas transition varieties affected fecundity and population parameters more.

In natural ecological systems, plants are often simultaneously attacked by both insects and pathogens, which can affect each other’s performance and the interactions can be extended to higher trophic levels, such as parasitoids. The English grain aphid (Sitobion avenae) and powdery mildew (Blumeria graminis f. sp. tritici) are two common antagonists that pose a serious threat to wheat production. Numerous studies have investigated the effect of a single factor (insect or pathogen) on wheat production. However, investigation on the interactions among insect pests, pathogens, and parasitoids within the wheat crop system are rare. Furthermore, the influence of the fungicide, propiconazole, has been found to imitate the natural ecosystem. Therefore, this study investigated the effects of B. graminis on the biological performance of grain aphids and the orientation behavior of its endoparasitic wasp Aphidius gifuensis in the wheat system. Our findings indicated that B. graminis infection suppressed the feeding behavior, adult and nymph weight, and fecundity and prolonged the developmental time of S. avenae. We found that wheat host plants had decreased proportions of essential amino acids and higher content of sucrose following aggravated B. graminis infection. The contents of Pro and Gln increased in the wheat plant tissues after B. graminis infection. In addition, B. graminis infection elicited immune responses in wheat: increase in the expression of defense genes, content of total phenolic compounds, and activity of three related antioxidant enzymes. Moreover, co-infection of B. graminis and S. avenae increased the attraction to A. gifuensis compare to that after infestation with aphids alone. In conclusion, our results indicated that B. graminis infection adversely affected the performance of S. avenae in wheat through restricted nutrition and induced defense response. Furthermore, the preference of parasitoids in such an interactive environment might provide an important basis for pest management control.

In nature, plants have evolved sophisticated defense mechanisms against the attack of pathogens and insect herbivores. Plant volatile-mediated plant-to-plant communication has been assessed in multitrophic systems in different plant species and different pest species. β-ocimene is recognized as an herbivore-induced plant volatile that play an important role in the chemical communication between plants and pests. However, it is still unclear whether β-ocimene can active the defense mechanism of Chinese cabbage Brassica pekinensis against the peach aphid Myzus persicae. In this study, we found that treatment of Chinese cabbage with β-ocimene inhibited the growth of M. persicae in terms of weight gain and reproduction. Moreover, β-ocimene treatment negatively influenced the feeding behavior of M. persicae by shortening the total feeding period and phloem ingestion and increasing the frequency of stylet puncture. When given a choice, winged aphids preferred to settle on healthy Chinese cabbage compared with β-ocimene-treated plants. In addition, performance of the parasitoid Aphidius gifuensis in terms of Y-tube olfaction and landings was better on β-ocimene-treated Chinese cabbage than on healthy plants. Furthermore, β-ocimene significantly increased the expression levels of salicylic acid and jasmonic acid marker genes and the accumulation of glucosinolates. Surprisingly, the transcriptional levels of detoxifying enzymes (CYP6CY3, CYP4, and GST) in aphids reared on β-ocimene-treated Chinese cabbage were significantly higher than those of aphids reared on healthy plants. In summary, our results indicated that β-ocimene can activate the defense response of Chinese cabbage against M. persicae, and that M. persicae can also adjust its detoxifying enzymes machinery to counter the host plant defense reaction.

Plant leaves of different ages differ in nutrients and toxic metabolites and thus exhibit various resistance levels against insect herbivores. However, little is known about the influence of leaf ontogeny on plant resistance to phloem-feeding insects. In this study, we found that the green peach aphid, Myzus persicae, preferred to settle on young cabbage leaves compared with mature or old leaves, although young leaves contained the highest concentration of glucosinolates. Furthermore, aphids feeding on young leaves had higher levels of glucosinolates in their body, but aphids performed better on young leaves in terms of body weight and population growth. Phloem sap of young leaves had higher amino acid:sugar molar ratio than mature leaves, and aphids feeding on young leaves showed two times longer phloem feeding time and five times more honeydew excretion than on other leaves. These results indicate that aphids acquired the highest amount of nutrients and defensive metabolites when feeding on young cabbage leaves that are strong natural plant sinks. Accordingly, we propose that aphids generally prefer to obtain more nutrition rather than avoiding host plant defense, and total amount of nutrition that aphids could obtain is significantly influenced by leaf ontogeny or source-sink status of feeding sites.

The behaviour of the greenhouse whitefly, Trialeurodes vaporariorum Westwood (Hemiptera: Aleyrodidae), is known to be affected by plant volatile cues, but its attraction or repellent to specific volatile cues has not been deeply studied yet. Therefore, the aim of our study was to identify the most attractive plant among cultivars of tomato (Solanum lycopersicum) and eggplant (Solanum melongena) to evaluate the volatiles of plants to identify the chemical compound(s) that attract T. vaporariorum. We speculated that whitefly–host plant interaction primarily depends on plant volatile emissions and that once the plant is damaged, it might attract more whiteflies. Three intact (uninfested) tomato, four intact eggplant cultivars and whitefly-infested plants of the most whitefly attractive tomato and eggplant cultivars were examined by behavioral assay experiments for attractiveness to T. vaporariorum and headspace volatile were determined by Solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS). Whiteflies had the highest preference for the intact eggplant Kuai Yuan Qie (KYQ) among the eggplant and the tomato plant cultivars in bioassay experiments. Although both male and female whiteflies were significantly more attracted to infested KYQ plants than to intact plants, whitefly females did not select conspecific-infested YG plants. The volatile emissions among different plant cultivars in individual species and infested versus intact plants were significantly different. Among these volatiles, identified major green leaf volatile (GLVs) ((Z)-3-hexen-1-ol) and terpenoids (α-pinene, (E)-β-caryophyllene, α-humulene, azulene) showed a constitutive relationship with the most whitefly preference plants. Our findings provide new insights into the chemical compounds that attract or repel whiteflies.

To calculate the volume of occult blood loss after unicompartment knee arthroplasty(UKA), and analyze its influential factors by comparing with total knee arthroplasty.A retrospective study of 130 cases from July 2012 to July 2015 were enrolled, of which and 65 cases were UKA, and 65 cases were TKA. In UKA group there were 27 males and 38 females, and the mean age was (62.3±4.3) years old (ranged, 50 to 82 years old). There were 14 cases older than 70 years old, while 51 cases among 50 to 70 years old. The mean Body mass index (BMI) was (23.9±2.6) kg/m²(ranged, 18.3 to 30.1 kg/m²). In TKA group there were 23 males and 42 females, and the mean age was (67.4±4.9) years old (ranged, 57 to 81 years old). There were 34 cases older than 70 years old, while 31 cases among 50 to 70 years old. The mean BMI was (25.6±2.3) kg/m²(ranged, 20.6 to 33.1 kg/m²). Hidden blood loss was calculated according to Gross equation, and the differences between the two groups including different ages and genders were observed subsequently. The differences of red blood cell change between two groups were observed dynamically postoperatively.Postoperative hidden blood loss was (375.25±168.09) ml, HSS score was 87.11±5.39 in UKA group. Hidden blood loss was (898.81±221.47) ml, HSS score was 82.23±3.08 in TKA group. The differences between the two groups were significant. There were no significant differences in ages and genders. There were no significant relationship between hidden blood loss and HSS score or BMI, while the differences of red blood cell change were significant on the second day, the 4th day and the 5th day. There was no allogenic blood transfusion in UKA group, while 3 patients with transfusion in TKA group.Hidden blood loss is part of total blood loss after UKA, and the volume in UKA is less than that in TKA. It could be compensated by the body and rarely affect the knee function. The hidden blood is not a risk factor of transfusion.

To analyze and compare the efficacy of surgical approaches and fixations of anterolateral approach, lateral approach and posterolateral approach in treating posterolateral tibial plateau fracture.A retrospective study of 44 cases from May 2010 to July 2014 were enrolled, of which there were 21 males and 23 females, and the mean age was 42.5 years old (ranged, 26 to 61 years). All the cases were divided into 3 groups according to the surgical approach, group A was anterolateral approach (19 cases), group B was lateral approach (15 cases), group C was posterolateral approach (10 cases). Operative time and bleeding volum were compared and the knee function was observed.The mean operative time of group A was (91.3±10.4) min, and the bleeding volum of which was (175.3±20.3) ml. The mean operative time of group B was(86.6±9.2) min, and the bleeding volum of which was(155.8±18.2) ml. The mean operative time of group C was (109.5±10.8) min, and the bleeding volum of which was(235.9±29.1) ml. There were significant differences in operative time and bleeding volum between group C and the other two groups(The operative time of posterolateral approach was longer than anterolateral approach or lateral approach, as well as the bleeding volum which was higher in posterolateral approach, while no significant difference of the knee function was observed in these 3 different approaches.

Tomato (Solanum lycopersicum) production is seriously hampered by the infestation of the sweetpotato whitefly, Bemisia tabaci MEAM 1 (Middle East-Asia Minor 1). The infestation behavior of the whiteflies could be affected by the quantity of plant released volatile organic compounds (VOCs) related to nitrogen concentrations of the plant. In this study, we determined the infestation behavior of B. tabaci to the tomato plants that produced different levels of VOCs after application of different levels of nitrogen with a wind tunnel and an olfactometer. We also analyzed the VOCs released from nitrogen-treated tomato plants using solid phase microextraction and gas chromatography-mass spectrometry. The results revealed that the production of eight VOCs (β-pinene, (+)-4-carene, α-terpinene, p-cymene, β-phellandrene, α-copaene, β-caryophyllene, and α-humulene) was reduced after the plants were treated with high levels of nitrogen. However, more whiteflies were attracted to the tomato plants treated with high levels of nitrogen than to the plants treated with normal or below normal levels of nitrogen. These results clearly indicated that nitrogen can change the quality and quantity of tomato plant volatile chemicals, which play important roles in B. tabaci host plant selection.

The green peach aphid, Myzus persicae Sulzer, is a notorious pest on vegetables, which often aggregates in high densities on crop leaves. In this study, we investigated whether M. persicae could suppress the resistance level of Chinese cabbage Brassica pekinensis. M. persicae performed better in terms of weight gain (~33% increase) and population growth (~110% increase) when feeding on previously infested (pre-infested) Chinese cabbage compared with those on non-infested plants. However, when given a choice, 64% of the aphids preferred to settle on non-infested leaves, while 29% of aphids chose pre-infested leaves that had a 2.9 times higher concentration of glucosinolates. Aphid feeding significantly enhanced the amino acid:sugar ratio of phloem sap and the absolute amino acid concentration in plant leaves. Aphid infestation significantly increased the expression levels of salicylic acid (SA) marker genes, while it had marginal effects on the expression of jasmonate marker genes. Exogenously applied SA or methyl jasmonate had no significant effects on M. persicae performance, although these chemicals increased glucosinolates concentration in plant leaves. M. persicae infestation increase amino acid:sugar ratio and activate plant defenses, but aphid performed better on pre-infested plants, suggesting that both nutrition and toxics should be considered in insect-plant interaction.

The behavior of the greenhouse whitefly, Trialeurodes vaporariorum Westwood (Hemiptera: Aleyrodidae), is known to be affected by plant volatile cues, but its attraction or repellent to specific volatile cues has not been deeply studied yet. Therefore, the aim of our study was to identify the most attractive plant among cultivars of tomato (Solanum lycopersicum) and eggplant (Solanum melongena) to evaluate the volatiles of plants to identify the chemical compound(s) that attract T. vaporariorum. We speculated that whitefly–host plant interaction primarily depends on plant volatile emissions and that once the plant is damaged, it might attract more whiteflies. Three intact (uninfested) tomato, four intact eggplant cultivars and whitefly infested plants of the most whitefly attractive tomato and eggplant cultivars were examined by behavioral assay experiments for attractiveness to T. vaporariorum and headspace volatile were determined by solid-phase microextraction (SPME) and gas chromatography-mass spectrometry. Whiteflies had the highest preference for the intact eggplant Kuai Yuan Qie (KYQ) among the eggplant and the tomato plant cultivars in bioassay experiments. Although both male and female whiteflies were significantly more attracted to infested KYQ plants than to intact plants, whitefly females did not select conspecific-infested YG plants. The volatile emissions among different plant cultivars in individual species and infested versus intact plants were significantly different. Among these volatiles, identified major green leaf volatiles [(Z)-3-hexen-1-ol] and terpenoids [α-pinene, (E)-β-caryophyllene, α-humulene, azulene] showed a constitutive relationship with the most whitefly preference plants. Our findings provide new insights into the chemical compounds that attract or repel whiteflies.

Plant leaves of different ages differ in nutrition and toxic metabolites and thus exhibit various resistance levels against insect herbivores. However, little is known about the relationship between leaf ontogeny and plant resistance against phloem-feeding insects. In this study, we found that the green peach aphid, Myzus persicae Sulzer, preferred to settle on young cabbage leaves (Brassica oleracea L. var. capitata) rather than mature or old leaves, although young leaves contained the highest concentration of glucosinolates. Furthermore, aphids feeding on young leaves had higher levels of glucosinolates in their bodies, but aphids performed better on young leaves in terms of body weight and population growth. The concentration of glutamine in young leaves was the highest, which stimulated aphids feeding when added to the sucrose solution. Phloem sap of young leaves had higher amino acid:sucrose molar ratio than mature leaves, and aphids feeding on young leaves showed two times longer phloem feeding time and five times more dry honeydew excretion than on other leaves. These results indicate that aphids acquired the highest amount of nutrition and defensive metabolites when feeding on young cabbage leaves that are strong natural plant sinks. The higher phloem sap availability of young leaves likely contributes to the attractiveness and suitability for aphids and may compensate the negative effects of glucosinolates on aphids. According to these findings, we propose that phloem sap availability influenced by leaf ontogeny and source-sink status play a significant role in plant-aphid interaction, which desires more attention in future research.

Tripterygium wilfordii Hook.f. (Celastraceae) produces a vast array of natural products, mainly terpenes and sesquiterpene pyridine alkaloids, which have strong biological activities. The limited gene information on the biosynthesis pathways restricts the metabolic engineering of this valuable natural resource. In this study, a suppression subtractive hybridization library of methyl jasmonate (MeJA)-elicited T. wilfordii Hook.f. adventitious root was successfully constructed. To clone the genes involved in the downstream of biosynthetic pathways, a long elicitation time of 12 days was applied. After evaluating the subtraction efficiency through polymerase chain reaction and reverse northern hybridization, 818 clones were randomly sequenced. Basic Local Alignment Search Tool (BLAST) analysis showed that 568 of the 735 successfully expressed sequence tags were annotated. Overrepresentation of gene ontology analysis indicated that the suppression subtractive hybridization library enriched genes that are related to biotic and abiotic stimuli. After BLAST and phylogenetic analyses, 11 genes were found to be putatively involved in the biosynthesis of terpene and sesquiterpene pyridine alkaloid, and nine transcription factors were putatively involved in modulation. The expression modulations of these promising genes were further examined after short MeJA treatment. This study provides valuable information for further research on the production improvement of terpene and sesquiterpene pyridine alkaloids through biotechnology methods.

Polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE), and a culture-dependent technique were used to study the diversity of the intestinal bacterial community in adult Dastarcus helophoroides (Fairmaire) (Coleoptera: Bothrideridae). Universal bacterial primers targeting 200 bp regions of the 16S rDNA gene were used in the PCR-DGGE assay, and 14 bright bands were obtained. The intestinal bacteria detected by PCR-DGGE were classified to Enterococcus (Lactobacillales: Enterococcaceae), Bacillus (Bacillales: Bacillaceae), Cellvibrio (Pseudomonadales: Pseudomonadaceae), Caulobacter (Caulobacterales: Caulobacteraceae), and uncultured bacteria, whereas those isolated by the culture-dependent technique belonged to Staphylococcus (Bacillales: Staphylococcaceae), Pectobacterium Enterobacteriales: Enterobacteriaceae), and Enterobacter (Enterobacteriales: Enterobacteriaceae). These intestinal bacteria represented the groups Lactobacillales ( Enterococcus ), Pseudomonadales ( Cellvibrio ), Caulobacterales ( Caulobacter ), Bacilli ( Bacillus and Staphylococcus ), and Gammaproteobacteria ( Pectobacterium and Enterobacter ). Our results demonstrated that PCR-DGGE analysis and the culture-dependent technique were useful in determining the intestinal bacteria of D. helophoroides and the two methods should be integrated to characterize the microbial community and diversity.

Aphids exhibit wing polyphenism (winged or wingless) for adaption to predictable or temporally heterogeneous environmental changes; however, the underlying mechanism is still unclear. This morphological change could be stimulated by high aphid density, which in turn could affect octopaminergic signaling in aphids. Octopamine is a neurotransmitter synthesized in insects that can modify their physiological metabolism, locomotion, and other behaviors. We designed experiments to determine whether octopamine functions in wing formation of the pea aphid, Acyrthosiphon pisum (Harris). We determined gene expression of tyramine β-hydroxylase (TβH), a key enzyme in octopamine synthesis at different developmental stages, in different body parts, and in different densities of aphids. We also used TβH RNAi, octopamine receptor agonists (octopamine and synephrine), and an antagonist (mianserin) to modify octopaminergic signaling. We found that transcription of TβH was related to aphid density, which affected the proportion of winged offspring. By manually modifying the mother’s octopaminergic signaling, TβH expression was suppressed, and TβH (enzyme) activity decreased. The proportion of winged offspring was also affected. Our results showed that octopamine could be a link in the wing determination system, as well as environmental stimulation. The RNAi results showed that the decrease of TβH expression increased aphid’s reproduction; however, the decrease of TβH expression declined the numbers of winged-offspring producers, but did not affect the proportion of winged nymphs produced by the winged-offspring producer. In conclusion, the decline in the proportion of winged daughters in the next generation was caused by the decline of winged nymph producers.

Tomato (Solanum lycopersicum) production is seriously hampered by the infestation of the sweetpotato whitefly, Bemisia tabaci MEAM 1 (Middle East-Asia Minor 1). The infestation behavior of the whiteflies could be affected by the quantity of plant released volatile organic compounds (VOCs) related to nitrogen concentrations of the plant. In this study, we determined the infestation behavior of B. tabaci to the tomato plants that produced different levels of VOCs after application of different levels of nitrogen with a wind tunnel and an olfactometer. We also analyzed the VOCs released from nitrogen-treated tomato plants using solid phase microextraction and gas chromatography-mass spectrometry. The results revealed that the production of eight VOCs (β-pinene, (+)-4-carene, α-terpinene, p-cymene, β-phellandrene, α-copaene, β-caryophyllene, and α-humulene) was reduced after the plants were treated with high levels of nitrogen. However, more whiteflies were attracted to the tomato plants treated with high levels of nitrogen than to the plants treated with normal or below normal levels of nitrogen. These results clearly indicated that nitrogen can change the quality and quantity of tomato plant volatile chemicals, which play important roles in B. tabaci host plant selection.

To study the correlation of postoperative femorotibial angle with medial compartmental joint line elevation after unicompartmental arthroplasty(UKA), as well as the correlation of joint line elevation with the clinical function by measuring radiological joint line.A retrospective study of 56 patients from July 2012 to August 2015 was performed. The mean body mass index (BMI) was 23.5 (ranged, 18.3 to 30.1). The standing anteroposterior radiographs of these patients were assessed both pre-and post-operatively, and the knee function was evaluated according to HSS grading. The correlation between postoperative femorotibial angle(FTA) and joint line elevation was analyzed as well as the correlation between joint line elevation and the clinical function.The mean medial joint line elevation was (2.2±2.0) mm(ranged, -3.3 to 7.0 mm), and the mean FTA correction was (2.3±3.0)°(ranged, -4.5° to 9.6°). The mean follow-up period was 12.2 months. There was a significant correlation between in joint line elevation and FTA correction(There was a significant correlation between medial compartmental joint line elevation and FTA correction after UKA, and the proximal tibial osteotomy was critical during the procedure. There was no significant correlation between joint line elevation and the clinical function, which may be related to the design of UKA prosthesis.

Wingless forms of aphids are relatively sedentary, and have a limited ability to migrate or disperse. However, they can drop off hosts or walk away if disturbed, or their food quality or quantity become deteriorated. Earlier, we found that the pea aphid, Acyrthosiphon pisum (Harris, 1776), could use differed strategies to escape danger and locate new host plants. To determine the mechanisms behind the different strategies, we undertook a series of studies including the aphids' host location, energy reserves under starvation, glycogenesis, sugar assimilation, olfactory and probing behaviors. We found that in our controlled laboratory conditions, one strain (local laboratory strain) moved longer distances and dispersed wider ranges, and correspondingly these aphids assimilated more sugars, synthesized more glycogen, and moved faster than another strain (collected from Gansu Province, northwestern China). However, the latter strain could locate the host faster, probed leaves more frequently, and identified plant leaves more accurately than the former strain after they were starved. Our results explained how flightless or wingless insects adapt to fit biotic and abiotic challenges in the complex processes of natural selection., Summary: Two strains of pea aphid (Acyrthosiphon pisum) show different dispersal strategies, glycogen level and probing behaviors. One strain [Yangling strain (YL)] moves longer and disperses wider than another strain [Lanzhou (LZ)] based on higher glycogen. LZ strain probed more frequently, and identified plant more accurately than YL strain.

To explore mid-term follow up results of distal fixation prosthesis in treating unstable intertrochanteric fractures in elderly patients.From May 2008 to March 2014,58 elderly patients with unstable intertrochanteric were treated with distal fixation prosthesis, among them, there were 15 males and 43 females aged from 75 to 87 years old with an average of 83.2 years old. Fracture were classified according to Evans classification, 39 cases were type I c and 19 cases were type I d. Surgical risk was evaluated before operation, 9 patients were performed total hip arthroplasty and 49 patients were performed prosthetic replacement hip joint function of patients with different age period, Evans classificaton, prothesis type, fixation method were evaluated respectively by using Harris score.Fifty-six patients were followed up from 13 to 36 months with an average of 21.6 months. Harris score was 83.51 ± 6.40, 5 cases got excellent results, 38 cases good and 13 cases moderate. Harris score of patients aged from 75 to 80 years old was 88.64 ± 2.35, 81.64 ± 6.40 in patients aged more than 80 years old, and had significant differences between two groups; Harris score in patients with type Evans I c was 83.64 ± 6.53, and 83.11 ± 6.08 in type Evans I d, while there was no significant differences between two groups. There was no obvious meaning in Harris score between patients with tension band (83.63 ± 6.15) and without tension band (82.41 ± 6.57). There was no significant meaning in Harris score between patients with normal distal fixation prosthesis (83.34 ± 6.43) and femoral moment reconstruction distal fixation prosthesis (83.92 ± 6.51). There was 1 patient occurred hip joint dislocation on the operative side and re-dislocation after manual reduction, then received open reduction. Two patients occurred femoral osteolysis without clinical symptoms, and treated conservative treatment.Artificial joint replacement for unstable intertrochanteric fractures in elderly patients, hip joint function in patients aged more than 80 years old is worse, while there was no obvious market effect in fracture classification, whether to use tension band and type of distal fixation prosthesis, moreover, proximal femoral osteolysis should be focused on.

Osteopetrosis is an inherited disorder characterized by increased bone density and brittle bone quality. Degenerative changes often occur after the age of 40 in patients with osteopetrosis. Operative intervention is the primary treatment option if the clinical manifestation of secondary osteoarthritis is severe. A 44-year-old male suffering autosomal dominant osteopetrosis and progressive unilateral hip osteoarthritis required a total hip arthroplasty. However, there were several technical challenges associated with this procedure including creating a femoral medullary canal and developing a Vancouver type B2 periprosthetic femoral fracture postoperatively. To afford some experience for the management of similar cases, we here present our technical solutions to these problems.

Terrestrial insects deposit a layer of hydrocarbons (HCs) as waterproofing agents on their epicuticle. The insect-specific CYP4G genes, subfamily members of P450, have been found in all insects with sequenced genomes to date. They are critical for HC biosynthesis in Drosophila; however, their functional roles in other insects including the piercing-sucking hemipterous aphids remain unknown. In this study, we presented the molecular characterization and a functional study of the CYP4G51 gene in the pea aphid, Acyrthosiphon pisum (Harris). CYP4G51 transcript was detectable across the whole life cycle of A. pisum, and was prominently expressed in the aphid head and abdominal cuticle. Up-regulation of CYP4G51 under desiccation stress was more significant in the third instar nymphs compared with the adults. Also, up-regulation of CYP4G51 was observed when the aphids fed on an artificial diet compared with those fed on the broad bean plant, and was positively correlated with a high level of cuticular HCs (CHCs). RNAi knockdown of CYP4G51 significantly reduced its expression and caused reductions in both internal and external HCs. A deficiency in CHCs resulted in aphids being more susceptible to desiccation, with increased mortality under desiccation stress. The current results confirm that CYP4G51 modulates HC biosynthesis to protect aphids from desiccation. Moreover, our data also indicate that saturated and straight-chain HCs play a major role in cuticular waterproofing in the pea aphid. A. pisum CYP4G51 could be considered as a novel RNAi target in the field of insect pest management.

Herbivores can ingest and store plant-synthesized toxic compounds in their bodies and sequester those compounds for their own benefits. The broad bean, Vicia faba L., contains a high quantity of L-DOPA (L-3,4-dihydroxyphenylalanine), which is toxic to many insects. However, the pea aphid, Acyrthosiphon pisum, can feed on V. faba normally, whereas many other aphid species could not. In this study, we investigated how A. pisum utilizes plant-derived L-DOPA for their own benefit. L-DOPA concentrations in V. faba and A. pisum were analyzed to prove L-DOPA sequestration. L-DOPA toxicity was bioassayed using an artificial diet containing high concentrations of L-DOPA. We found that A. pisum could effectively adapt and store L-DOPA, transmit it from one generation to the next. We also found that L-DOPA sequestration verity differed in different morphs of A. pisum. After analyzing the melanization efficiency in wounds, mortality and deformity of the aphids at different concentrations of L-DOPA under ultraviolet radiation (UVA 365.0 nm for 30 min), we found that A. pisum could enhance L-DOPA assimilation for wound healing and UVA-radiation protection. Therefore, we conclude that A. pisum could acquire L-DOPA and use it to prevent UVA damage. This study reveals a successful co-evolution between A. pisum and V. faba.