Construction of a redox pathway through a polyoxometalate and covalent organic framework for H2O2 photosynthesis.

A novel type of electron donor-acceptor system was built from a nitrogen-rich covalent organic framework (PC) and a polyoxometalate (BW12), fabricating a composite material (BW12@PC-250), which shows significantly improved photocatalytic H2O2 yield (56.4 µM h-1) under full spectrum illumination in pure water, being about 30 times higher than that of PC. This is due to the opening of the electron and proton transport pathway between PC and BW12, which paves a new way for POMs to modulate the photocatalytic reactions of COFs.

One-Step Green Synthesis of Isoeugenol Methyl Ether from Eugenol by Dimethyl Carbonate and Phase-Transfer Catalysts.

In this paper, the green synthesis of isoeugenol methyl ether (IEME) from eugenol by O-methylation and isomerization is completed using a one-step green process. In the methylation reaction, dimethyl carbonate (DMC) was used as a green chemistry reagent instead of the traditional harmful methylation reagents, in accordance with the current concept of green chemistry. The phase transfer catalyst (PTC) polyethylene glycol 800 (PEG-800) was introduced into the isomerization reaction to break the barrier of difficult contact between solid and liquid phases and drastically reduce the reaction conditions by shortening the reaction time and reducing the alkalinity of the reaction system. The catalytic systems for the one-step green synthesis of IEME were screened, and it was shown that the catalytic system "K2CO3 + PEG-800" was the most effective. The effects of reaction temperature, n(DMC):n(eugenol) ratio, n(PEG-800):n(eugenol) ratio, and n(K2CO3):n(eugenol) ratio on eugenol conversion, IEME yield, and IEME selectivity were investigated. The results showed that the best reaction was achieved at a reaction temperature of 140 °C, a reaction time of 3 h, a DMC drip rate of 0.09 mL/min, and n(eugenol):n(DMC):n(K2CO3):n(PEG-800) = 1:3:0.09:0.08. As a result of the conversion of 93.1% of eugenol to IEME, a yield of 86.1% IEME as well as 91.6% IEME selectivity were obtained.

Modified interproximal tunneling technique with customized sub-epithelial connective tissue graft for gingival papilla reconstruction: report of three cases with a cutback incision on the palatal side.

BACKGROUND: Gingival papilla defects, which cause an unpleasant appearance and involve the upper anterior teeth, may be triggered by several factors. Several noninvasive and invasive techniques have been proposed for gingival papilla reconstruction. The combination of interproximal tunneling and customized connective tissue grafts (CTGs) has shown promise in papilla augmentation. However, due to the narrowness and limited blood supply of the gingival papilla, the long-term outcomes of these techniques remain unpredictable. Therefore, achieving tension-free coronal advancement of the interdental papilla and proper placement of the CTG is crucial for successful long-term outcomes and could provide widely applicable methods for papilla augmentation. CASE REPORT: In this study, we enrolled three patients with gingival papilla defects in the maxillary anterior teeth. For reconstruction, we proposed a modified interproximal tunneling (MIPT) technique combined with a CTG. A crucial modification based on previous studies involved adding a cutback incision to the base of the palatal vertical incision, resulting in tension-free healing. Additionally, the CTG was sutured upright to further enhance the height of the gingiva papilla. To evaluate the efficacy of the MIPT technique, the clinical parameters-including the Jemt papilla index and the distance from the tip of the papilla to the interproximal contact point-were examined using a periodontal probe (UNC15, Hu-friedy) at baseline and 12 months after surgery. All three patients achieved satisfactory papilla reconstruction 12 months after the surgery. These three cases were used to evaluate the efficacy of the MIPT technique combined with the customized CTG. An average increase in the Jemt papilla score from 1.6 to 2.8 and a reduction in the distance from the papilla tip to the contact point of adjacent teeth from 2 mm to 0.08 mm were observed 12 months after surgery. CONCLUSION: The preliminary results confirmed that this technique holds promise for gingival papilla augmentation between tooth/tooth or tooth/implant.

Ileus induced by the combination of vinca alkaloids and posaconazole in a patient with acute lymphoblastic leukemia: a case report and literature review.

Vinca alkaloid (VA)-induced ileus, a rare but severe autonomic neuropathy, can be enhanced by concomitant use of antifungal triazole agents. We herein present a case of VA-induced ileus in a 17-year-old girl who was diagnosed with B-cell acute lymphoblastic leukemia. On day 1, the patient received cyclophosphamide, vincristine, and methylprednisolone. On day 2, she began treatment with posaconazole oral suspension at 200 mg three times daily for prophylaxis against invasive fungal infection. On day 5, she began induction therapy consisting of vindesine, methylprednisolone, daunorubicin, and cyclophosphamide. The patient developed severe abdominal pain with marked constipation on day 11 and was diagnosed with incomplete ileus. After switching the antifungal agent to micafungin, performing gastrointestinal decompression, administering parenteral nutrition, and omitting the fourth dose of vindesine, the ileus symptoms were relieved. This case emphasizes the potential interaction between VAs and posaconazole. We also herein present a review of the literature on ileus caused by the combination of VAs and antifungal triazole agents. In clinical practice, physicians and pharmacists should be aware of the possibility of ileus caused by the use of VAs in combination with posaconazole. It is important to reduce complications during chemotherapy to improve patients' prognosis.

The risk factors of biliary fistula after radical resection of perihilar cholangiocarcinoma in elderly patients and its influence on prognosis: a retrospective cohort study.

Background: Nowadays, the incidence of perihilar cholangiocarcinoma (PHCC) is increasing yearly, and biliary fistula is a common complication. However, there are few reports on the factors affecting the occurrence of biliary fistula. This study aimed to analyze the risk factors for the occurrence of biliary fistula after radical surgery for PHCC in elderly patients to provide a reference basis for improving the prognosis. Methods: From April 2016 to April 2021, we randomly included 250 elderly patients with pathologically diagnosed PHCC. Based on our established inclusion and exclusion criteria, we included 211 patients finally Bile drainage for 3 consecutive days after abdominal drainage or single bile drainage ≥100 mL/d was used as the diagnostic criteria for biliary fistula. Multiple logistic regression was used to analyze the independent risk factors for biliary fistula after radical surgery for PHCC. Besides, the Prognostic Score (PS), the Karnofsky performance score (KPS), and the quality of life (QOL) score were used to assess the patients' postoperative recovery, and survival curves were drawn to reflect their 1-year survival rate, using overall survival. Results: The statistical results showed a 36.5% incidence of biliary fistula. Our study showed that preoperative cholangitis, number of biliary anastomoses, etc. were independent risk factors for grade B and grade C biliary fistula. Besides, the presence of intraoperative hemorrhage (OR =0.223, P=0.006) and γ-glutamyl transpeptidase (γ-GT) on the first postoperative day (OR =1.011, P=0.013) were still independent risk factors for grade B biliary fistula, while C-reactive protein (CRP) (OR =1.026, P=0.011) and total bilirubin (TBil) (OR =0.003, P=1.066) on the first postoperative day and bile duct diameter (OR =0.299, P=0.020) were independent risk factors for grade C biliary fistula. Analysis of variance (ANOVA) showed statistically significant differences (P<0.05) between the three groups in terms of PS (P=0.000), KPS (P=0.001), and QOL scores (P=0.000). Conclusions: The incidence of postoperative biliary fistula remains high in elderly patients treated with radical surgery for PHCC and seriously affects their prognosis. Therefore, focusing on the clinical characteristics and risk factors of patients, improving surgical precision and enhancing postoperative patient care to reduce the likelihood of postoperative biliary fistula, thereby prolonging patient life.

Rhein induces bone regeneration via alleviating inflammation in murine periodontitis model.

OBJECTIVE: To evaluate the effect of rhein on eliminating the inflammation and promoting bone regeneration of periodontitis after local administration. MATERIALS AND METHODS: In vivo, periodontitis model was established in murine mandibular first molar by using ligature for 7 days, followed by ligature removal and local administration of rhein/vehicle for 7 consecutive days. In vitro, periodontal ligament fibroblasts were treated by LPS, along with the applications of rhein/vehicle. Histology and molecular biology approaches were applied for analysis. RESULTS: In vivo, rhein alleviated periodontitis inflammation through downregulating the inflammatory index and promoted the osteogenic potential of PDL fibroblasts in a dosage-dependent manner. The result of micro-CT validated this phenomenon. In vitro, rhein administration inhibited the phosphorylation and nuclear translocation of P65, along with the arose runx2 level of PDL fibroblasts with the stimulus of LPS in mimicking periodontitis. CONCLUSION: Rhein played its inhibitory role on inflammation via curbing the activation of P65 but uprising the activities of Runx2 in PDL fibroblasts in periodontitis microenvironment. These data suggested that rhein could be an effective and potential clinical choice for the treatment of periodontitis.

Geochemical characteristics and provenance of the detrital sediments in the junction area of Yinggehai and Qiongdongnan basins, South China Sea.

Together, the Yinggehai and Qiongdongnan basins have received a large amount of terrigenous sediments, but the provenance evolution of Cenozoic sediments in the two basins remains disputable. Combined with previous studies in the Yinggehai and Qiongdongnan basins, the elemental geochemistry of Oligocene to Pliocene sediment samples in the junction area of the two basins were analyzed to explore the tectonic implications, parent rock characteristics, and provenance evolution of the two basins during the Cenozoic. The results reveal that all the sediment samples were derived from continental island arc to passive continental margin settings. The light REE enrichment and stable content of heavy REE with large negative Eu anomalies indicate that they were probably derived from Hainan Island. The reconstructed provenance evolution model showed that the Red River Source (RRS) provided sedimentary materials for the Central Depression of Yinggehai Basin from the Oligocene to the Pliocene, and Hainan Island Source (HIS) was also one of the sources for sediments deposited in the Central Depression of Yinggehai Basin during the Miocene. However, most of the sediments preserved in the Yingdong Slope and Qiongdongnan Basin were derived from the HIS from the Oligocene to the Pliocene, and sediments deposited in the Yingdong Slope were also derived from the RRS during the Miocene. Furthermore, the junction area of the two basins had a mixed source of the RRS and HIS during the Cenozoic.

Transcriptional regulation of host insulin signaling pathway genes controlling larval development by Microplitis manilae parasitization.

Idiobiont parasitoids using other insects as hosts sabotage the host growth and development to ensure their offspring survival. Numerous studies have discovered that insect development is subtly regulated by the conserved insulin signaling pathway. However, little is known about how wasp parasitization disrupts host development controlled by the insulin signaling pathway. Here we address this study to determine the effect of wasp parasitism on host Spodoptera frugiperda development using the idiobiont parasitoid Microplitis manilae as a model. Upon M. manilae parasitization, the body weight, body length, and food consumption of host insect were dramatically reduced compared to the unparasitized S. frugiperda. We next identified the core genes involved in host insulin signaling pathway and further analyzed the domain organizations of these genes. Phylogenetic reconstruction based on the insulin receptors clustered S. frugiperda together with other noctuidae insects. In the latter study, we profiled the expression patterns of host insulin signaling pathway genes in response to M. manilae parasitization at 2, 24, and 48 h, significant decreases in mRNA levels were recorded in S. frugiperda larvae upon 24 and 48 h parasitization. These current findings substantially add to our understanding of the physiological interaction between parasitoid and host insects, thus contributing to revealing the molecular mechanism of parasitic wasps regulating host development.

Molecular and Pharmacological Characterization of ß-Adrenergic-like Octopamine Receptors in the Endoparasitoid Cotesia chilonis (Hymenoptera: Braconidae).

Octopamine (OA) is structurally and functionally similar to adrenaline/noradrenaline in vertebrates, and OA modulates diverse physiological and behavioral processes in invertebrates. OA exerts its actions by binding to specific octopamine receptors (OARs). Functional and pharmacological characterization of OARs have been investigated in several insects. However, the literature on OARs is scarce for parasitoids. Here we cloned three ß-adrenergic-like OARs (CcOctßRs) from Cotesia chilonis. CcOctßRs share high similarity with their own orthologous receptors. The transcript levels of CcOctßRs were varied in different tissues. When heterologously expressed in CHO-K1 cells, CcOctßRs induced cAMP production, and were dose-dependently activated by OA, TA and putative octopaminergic agonists. Their activities were inhibited by potential antagonists and were most efficiently blocked by epinastine. Our study offers important information about the molecular and pharmacological properties of ß-adrenergic-like OARs from C. chilonis that will provide the basis to reveal the contribution of individual receptors to the physiological processes and behaviors in parasitoids.

An endoparasitoid uses its egg surface proteins to regulate its host immune response.

With proteomic analysis, we identified 379 egg surface proteins from an endoparasitoid, Cotesia chilonis. Proteins containing conserved enzymatic domains constitute a large proportion of egg surface components. Some proteins, such as superoxidase dismutase, homolog of C. rubecula 32-kDa protein, and immunoevasive protein-2A, are classical parasitism factors that have known functions in host immunity regulation. Melanization assays revealed that a novel egg surface protein, C. chilonis egg surface serpin domain-containing protein had the same function as a C. chilonis venom serpin, as both suppressed host melanization in a dose-dependent manner. C. chilonis egg surface serpin domain-containing protein is mainly transcribed in C. chilonis oocytes with follicular cells, and it is located on both the anterior and posterior sides of the mature egg surface. Additionally, we used LC-MS/MS to identify 586 binding proteins sourced from C. suppressalis plasma located on the eggshell surface of C. chilonis, which included some immunity-related proteins. These results not only indicate that C. chilonis uses its egg surface proteins to reduce the immune response of its host but also imply that endoparasitoid egg surface proteins might be a new parasitism factor involved in host immune regulation.

Selective Chemical Labeling and Sequencing of 5-Hydroxymethylcytosine in DNA at Single-Base Resolution.

5-Hydroxymethylcytosine (5hmC), the oxidative product of 5-methylcytosine (5mC) catalyzed by ten-eleven translocation enzymes, plays an important role in many biological processes as an epigenetic mediator. Prior studies have shown that 5hmC can be selectively labeled with chemically modified glucose moieties and enriched using click chemistry with biotin affinity approaches. Besides, DNA deaminases of the AID/APOBEC family can discriminate modified 5hmC bases from cytosine (C) or 5mC. Herein, we developed a method based on embryonic stem cell (ESC) whole-genome analysis, which could enrich 5hmC-containing DNA by selective chemical labeling and locate 5hmC sites at single-base resolution with enzyme-based deamination. The combination experimental design is an extension of previous methods, and we hope that this cost-effective single-base resolution 5hmC sequencing method could be used to promote the mechanism and diagnosis research of 5hmC.

Interferon Gamma-Inducible Protein 16 of Peripheral Blood Mononuclear Cells May Sense Hepatitis B Virus Infection and Regulate the Antiviral Immunity.

Interferon gamma-inducible protein 16 (IFI16) is a DNA sensor protein, which triggers interferon-beta (IFN-ß) production. However, the role of IFI16 in the innate immunity against hepatitis B virus (HBV) remains controversial. Peripheral blood mononuclear cells (PBMCs) and serum specimens were collected from 20 patients with chronic hepatitis B (CHB) receiving Peg-IFN-α2b therapy. IFI16 mRNA/protein of PBMCs and serum IFI16 at baseline and changes during Peg-IFN-α2b treatment were detected. The interaction between IFI16 and HBV DNA in the PBMCs was analyzed using chromatin immunoprecipitation assay. Leukemic T cell line CEM-C7 and HBV-replicating HepG2.2.15 cells were used to test the effects of interferon treatment and HBV replication on IFI16 expression. Compared with healthy controls, lower levels of IFI16 mRNA but more significant expression of IFI16 protein with heterogeneous degradation were detected in PBMCs of CHB patients. Early changes in IFI16 mRNA, but not IFNB mRNA of PBMCs or serum IFI16, were correlated to HBeAg seroconversion of Peg-IFN-α2b therapy. An interaction between IFI16 and HBV DNA was detected in the PBMCs. In the cultured HepG2.2.15 and CEM-C7 cells, interferons resulted in the translocalization of IFI16 from the cytoplasm to the nucleus and inhibited IFI16 degradation. IFI16 of PBMCs may play a role in sensing HBV infection, and early change in IFI16 mRNA of PBMCs is valuable to predict HBeAg seroconversion in Peg-IFN-α2b treatment. The influences on IFI16 degradation and subcellular location may present a molecular mechanism of antiviral activity of interferon.

Association between tooth loss rate and risk of mild cognitive impairment in older adults: a population-based longitudinal study.

Mild cognitive impairment (MCI) is a symptomatic predementia phase of the trajectory of cognitive decline, and its prevalence increases with age. Although the relationship between oral health and MCI have been explored previously, it is uncertain whether individuals with different tooth loss rates have altered MCI risks. We hereby conducted a longitudinal study by using data from the Chinese Longitudinal Healthy Longevity Survey to investigate the association. Tooth loss rate was defined as the difference of teeth between two interview waves divided by years of interval; participants were then grouped into four categories: stable, no tooth loss; mild, 0-1 tooth loss; middle, 1-2 tooth loss; and severe, more than 2 tooth loss per year. Cognitive function was assessed by the Chinese version of Mini-Mental State Examination. We used the generalized estimating equation model to estimate the odds ratio (OR) and the 95% confidence intervals (CIs) and applied the restricted cubic spline function to explore the dose-response association. Among 11,862 participants, 3,966 developed MCI in a median follow-up time of 5.93 years. Higher tooth loss rate was associated with an increased risk of MCI in elderly subjects. Compared with subjects with stable tooth, the corresponding ORs (95% CIs) were 0.94 (0.85-1.03), 1.16 (1.04-1.29) and 1.28 (1.17-1.40) for subjects with the mild, middle and severe rate of tooth loss. A nonlinear dose-response relationship was detected (Pnon-linearity = 0.0165). Similar results were observed in the subgroup analyses stratified by sex, age at baseline, and number of teeth at baseline. The positive association was only observed among denture nonwearers (OR middle vs stable: 1.19; 1.06-1.35; OR severe vs stable: 1.35; 1.22-1.50), but not among denture wearers. In conclusion, among elderly population in China, higher rate of tooth loss may be associated with an increased risk of MCI, while denture wearers may be less likely to develop MCI.

Cellular and humoral immune interactions between Drosophila and its parasitoids.

The immune interactions occurring between parasitoids and their host insects, especially in Drosophila-wasp models, have long been the research focus of insect immunology and parasitology. Parasitoid infestation in Drosophila is counteracted by its multiple natural immune defense systems, which include cellular and humoral immunity. Occurring in the hemocoel, cellular immune responses involve the proliferation, differentiation, migration and spreading of host hemocytes and parasitoid encapsulation by them. Contrastingly, humoral immune responses rely more heavily on melanization and on the Toll, Imd and Jak/Stat immune pathways associated with antimicrobial peptides along with stress factors. On the wasps' side, successful development is achieved by introducing various virulence factors to counteract immune responses of Drosophila. Some or all of these factors manipulate the host's immunity for successful parasitism. Here we review current knowledge of the cellular and humoral immune interactions between Drosophila and its parasitoids, focusing on the defense mechanisms used by Drosophila and the strategies evolved by parasitic wasps to outwit it.

Effects of transcutaneous electrical acupuncture point stimulation on peripheral capillary oxygen saturation in elderly patients undergoing colonoscopy with sedation: a prospective randomized controlled trial.

INTRODUCTION: This study investigated whether transcutaneous electrical acupuncture point stimulation (TEAS) at PC6 can reduce the proportion of elderly patients experiencing a drop of ⩾4% in peripheral capillary oxygen saturation (SpO2) while undergoing colonoscopy under sedation. METHODS: A total of 32 elderly patients (aged ⩾ 65 years) scheduled for colonoscopy were randomly assigned in a 1:1 ratio to receive either real or sham TEAS (treatment or control groups, respectively). Each patient received oxygen (2 L/min) delivered routinely via nasal cannula. The treatment group was given TEAS at PC6 for 20 min at 2 Hz frequency and 6 mA intensity; the control group underwent the same procedures but with zero frequency/intensity. SpO2 and other physiological parameters were measured prior to sedation and colonoscopy (baseline) and at seven other timepoints through departure from recovery. Depth of anesthesia was measured using a Narcotrend monitor. RESULTS: Significantly fewer patients in the treatment group experienced a ⩾4% decrease from baseline SpO2 (2/16) than patients in the control group (10/16; p = 0.004). The two groups were comparable with regard to respiratory rate, systolic and diastolic blood pressures, mean arterial pressure, and heart rate. CONCLUSION: TEAS applied at PC6 with 2 Hz frequency was feasible and may be helpful in reducing the rate of hypoxia in elderly patients during colonoscopy.Trial registration number: NCT03775122 (ClinicalTrials.gov).

ACE2 and TMPRSS2 Expression in Hepatocytes of Chronic HBV Infection Patients.

Background: Pre-existing liver disease is a risk factor for the worse prognosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. We aimed to evaluate whether chronic hepatitis B (CHB) and hepatocellular carcinoma (HCC) affect the expression of viral receptor angiotensin-converting enzyme 2 (ACE2) and transmembrane serine protease 2 (TMPRSS2) in the liver. Methods: Twelve pairs of matched liver tissues of HCC and para-carcinoma were collected from the First Affiliated Hospital of Zhejiang University School of Medicine. And 20 liver biopsies from CHB patients were collected from Peking University People's Hospital. The expression of ACE2 and TMRPSS2 were detected using immunofluorescence staining, western blot, and RT-qPCR. The effects of hepatitis B virus (HBV) replication or interferon on ACE2 and TMPRSS2 expression were tested in hepatic cell lines. Results: The mRNA expression of TMPRSS2 in HCC tissues was six-fold higher than that of para-carcinoma tissues (P = 0.002), whereas that of ACE2 was not statistically different between HCC and para-carcinoma tissues. Hepatocellular ACE2 expression was detected in 35% (7/20) of CHB patients and mostly distributed in the inflammatory areas. However, there was no difference in TMPRSS2 expression between areas with or without inflammation. IFN-α2b slightly induced ACE2 expression (2.4-fold, P = 0.033) in HepG2 cells but not in Huh-7, QSG-7701, and L-02 cells. IFN-α2b did not affect TMPRSS2 expression in these cell lines. In addition, HBV replication did not alter ACE2 expression in HepAD38 cells. Conclusions: Although HBV replication does not directly affect the expression of ACE2 and TMPRSS2, intrahepatic inflammation and carcinogenesis may increase their expression in some patients, which, in turn, may facilitate SARS-CoV-2 infection in hepatocytes.

A venom protein, Kazal-type serine protease inhibitor, of ectoparasitoid Pachycrepoideus vindemiae inhibits the hemolymph melanization of host Drosophila melanogaster.

Parasitic wasps inject various virulence factors into the host insects while laying eggs, among which the venom proteins, one of the key players in host insect/parasitoid relationships, act in host cellular and humoral immune regulation to ensure successful development of wasp progeny. Although the investigations into actions of venom proteins are relatively ample in larval parasitoids, their regulatory mechanisms have not been thoroughly understood in pupal parasitoids. Here, we identified a venom protein, Kazal-type serine protease inhibitor, in the pupal ectoparasitoid Pachycrepoideus vindemiae (PvKazal). Sequence analysis revealed that PvKazal is packed by a signal peptide and a highly conserved "Kazal" domain. Quantitative polymerase chain reaction analysis recorded a higher transcript level of PvKazal in the venom apparatus relative to that in the carcass, and the PvKazal messenger RNA level appeared to reach a peak on day 5 posteclosion. Recombinant PvKazal strongly inhibited the hemolymph melanization of host Drosophila melanogaster. Additionally, the heterologous expression of PvKazal in transgenic Drosophila reduced the crystal cell numbers and blocked the melanization of host pupal hemolymph. Our present work underlying the roles of PvKazal undoubtedly increases the understanding of venom-mediated host-parasitoid crosstalk.

Identification, Characterization and Expression Analysis of TRP Channel Genes in the Vegetable Pest, Pieris rapae.

Transient receptor potential (TRP) channels are critical for insects to detect environmental stimuli and regulate homeostasis. Moreover, this superfamily has become potential molecular targets for insecticides or repellents. Pieris rapae is one of the most common and widely spread pests of Brassicaceae plants. Therefore, it is necessary to study TRP channels (TRPs) in P. rapae. In this study, we identified 14 TRPs in P. rapae, including two Water witch (Wtrw) genes. By contrast, only one Wtrw gene exists in Drosophila and functions in hygrosensation. We also found splice isoforms of Pyrexia (Pyx), TRPgamma (TRPγ) and TRP-Melastatin (TRPM). These three genes are related to temperature and gravity sensation, fine motor control, homeostasis regulation of Mg2+ and Zn2+ in Drosophila, respectively. Evolutionary analysis showed that the TRPs of P. rapae were well clustered into their own subfamilies. Real-time quantitative PCR (qPCR) showed that PrTRPs were widely distributed in the external sensory organs, including antennae, mouthparts, legs, wings and in the internal physiological organs, including brains, fat bodies, guts, Malpighian tubules, ovaries, as well as testis. Our study established a solid foundation for functional studies of TRP channels in P. rapae, and would be benefit to developing new approaches to control P. rapae targeting these important ion channels.

Molecular and pharmacological characterization of a ß-adrenergic-like octopamine receptor from the green rice leafhopper Nephotettix cincticeps.

As the counterparts of noradrenaline and adrenaline in vertebrates, octopamine (OA) regulates multiple physiological and behavioral processes in invertebrate. OA mediates its effects via binding to specific octopamine receptors (OARs). Functional and pharmacological characterization of OARs have been reported in several insects. However, little work was documented in hemipteran insects. We cloned a ß-adrenergic-like OAR (NcOA2B2) from Nephotettix cincticeps. NcOA2B2 shares high similarity with members of the OA2B2 receptor class. Transcript level of NcOA2B2 varied in various tissues and was highly expressed in the leg. After heterologous expression in CHO-K1 cells, NcOA2B2 was dose-dependently activated by OA (EC50 = 2.56 nM) and tyramine (TA) (EC50 = 149 nM). Besides putative octopaminergic agonists, dopaminergic agonists and amitraz and DPMF potently activated NcOA2B2 in a dose-dependent manner. Receptor activity was blocked by potential antagonists and was most efficiently antagonized by asenapine. Phentolamine showed both antagonist and agonist effects on NcOA2B2. Our results offer the important information about molecular and pharmacological characterization of an OAR from N. cincticeps that will provide the basis for forthcoming studies on its roles in physiological processes and behaviors, and facilitate the design of novel insecticides for pest control.

Identification and Comparative Analysis of Venom Proteins in a Pupal Ectoparasitoid, Pachycrepoideus vindemmiae.

Parasitoid wasps inject venom containing complex bioactive compounds to regulate the immune response and development of host arthropods and sometime paralyze host arthropods. Although extensive studies have been conducted on the identification of venom proteins in larval parasitoids, relatively few studies have examined the pupal parasitoids. In our current study, a combination of transcriptomic and proteomic methods was used to identify 64 putative venom proteins from Pachycrepoideus vindemmiae, an ectoparasitoid of Drosophila. Expression analysis revealed that 20 tested venom proteins have 419-fold higher mean expression in the venom apparatus than in other wasp tissues, indicating their specialization to venom. Comparisons of venom proteins from P. vindemmiae and other five species spanning three parasitoid families detected a core set of "ancient" orthologs in Pteromalidae. Thirty-five venom proteins of P. vindemmiae were assigned to the orthologous groups by reciprocal best matches with venoms of other pteromalids, while the remaining 29 were not. Of the 35 categories, twenty-seven have orthologous relationships with Nasonia vitripennis venom proteins and 25 with venoms of Pteromalus puparum. More distant relationships detected that five and two venom proteins of P. vindemmiae are orthologous with venoms of two Figitidae parasitoids and a Braconidae representative, respectively. Moreover, twenty-two venoms unique to P. vindemmiae were also detected, indicating considerable interspecific variation of venom proteins in parasitoids. Phylogenetic reconstruction based on a set of single-copy genes clustered P. vindemmiae with P. puparum, N. vitripennis, and other members of the family Pteromalidae. These findings provide strong evidence that P. vindemmiae venom proteins are well positioned for future functional and evolutionary studies.