The Sequence Characteristics and Binding Properties of the Odorant-Binding Protein 2 of Euplatypus parallelus to Semiochemicals.

Euplatypus parallelus is one of the dominant rubber bark beetle species in Hainan's rubber-planting area. Semiochemicals, including the volatiles found in rubber trees and aggregation pheromones, play an important role in the search for suitable host plants. To examine the possible functional role of highly expressed odorant-binding protein 2 of Euplatypus parallelus (EparOBP2) in the semiochemical recognition process, we cloned and analyzed the cDNA sequence of EparOBP2. The results showed that EparOBP2 contains an open reading frame (ORF) of 393 bp that encodes 130 amino acids, including a 21-amino-acid residue signal peptide at the N-terminus. The matured EparOBP2 protein consists of seven α-helices, creating an open binding pocket and three disulfide bridges. The results of the fluorescence binding assay showed that EparOBP2 had high binding ability with α-pinene and myrcene. The docking results confirmed that the interactions of α-pinene and myrcene with EparOBP2 were primarily achieved through hydrophobic interactions. This study provides evidence that EparOBP2 may be involved in the chemoreception of semiochemicals and that it can successfully contribute to the integrated management of E. parallelus.

Efficacy of moxibustion in diarrhea-predominant irritable bowel syndrome model rats: a systematic review and meta-analysis.

Objective: To systematically evaluate the efficacy of moxibustion in diarrhea-predominant irritable bowel syndrome (IBS-D) model rats. Methods: A comprehensive search was conducted in the China National Knowledge Infrastructure, WanFang Data, VIP, PubMed, Embase, and Web of Science databases from their inception to June 30, 2023. Relevant animal experiments investigating moxibustion for treating IBS-D in model rats were included. Two independent researchers screened the literature, extracted data, and evaluated the risk of bias in the selected studies. The meta-analysis was performed using RevMan 5.3 software. Results: In total, 21 animal studies comprising 680 model rats were included. The meta-analysis results demonstrated that moxibustion enhanced the threshold capacity of the abdominal withdrawal reflex (AWR) [standardized mean difference (SMD) = 1.84; 95% confidence interval (CI): 1.09, 2.60; p < 0.00001], ameliorated the rate of loose stool (SMD = -4.03; 95% CI: -5.76, -2.30; p < 0.00001), and decreased the colon 5-hydroxytryptamine (SMD = -3.67; 95% CI: -5.33, -2.01; p < 0.00001), serum interleukin-1ß (SMD = -3.24, 95% CI: -4.06, -2.41; p < 0.00001), serum tumor necrosis factor-α (SMD = -2.35, 95% CI: -4.12, -0.58; p < 0.00001), and serum substance P (SMD = -5.14, 95% CI: -8.45, -1.83; p = 0.002) concentrations. Moxibustion did not affect the blood calcitonin gene-related peptide level compared to the blank model group (p = 0.15). Conclusion: Moxibustion modulated the brain-gut interaction, reduced visceral hypersensitivity, inhibited intestinal inflammation, and regulated the immune balance, improving the rate of loose stool and increasing the AWR threshold capacity in IBS-D model rats, achieving good analgesic and antidiarrheal effects. However, these conclusions require further validation due to limitations in the quantity and quality of the included studies.

Ligand-binding properties of XaffOBP9, a Minus-C odorant-binding protein from Xyleborus affinis (Coleoptera: Curculionidae: Scolytinae).

Xyleborus affinis, one of the most important pests of rubber trees, has caused severe damage to the natural rubber industry in Hainan province. The ability to detect host plants through a sensitive and specific olfactory system is crucial for Xyleborus affinis. Odorant binding proteins (OBPs) are believed to bind and carry hydrophobic active compounds from the environment to the surface of olfactory receptor neurons. To investigate the potential functional role of the highly expressed XaffOBP9 in binding with semiochemicals, we cloned and analyzed the cDNA sequence of XaffOBP9. The results showed that XaffOBP9 contains a 411bp open reading frame that encodes 136 amino acids. Then XaffOBP9 was expressed in Escherichia coli. The binding affinity of the recombinant OBP to 15 different ligands (14 host plant volatiles and 1 aggregation pheromone) was then examined using a fluorescence competitive binding approach. The results demonstrated that XaffOBP9 exhibited broad binding capabilities and strong affinities for 14 ligands. The structure of XaffOBP9 and its interactions with fourteen ligands were further analyzed by modeling and molecular docking, respectively. Based on the docking result, we found hydrophobic interactions are important between XaffOBP9 to these ligands and three amino acid residues (L71, Y106, and L114) were highly overlapped and contributed to the interaction with ligands. Mutation functional assays confirmed that the mutant L114A showed significantly reduced binding capacity to these ligands. This study suggested that XaffOBP9 may be involved in the chemoreception of semiochemicals and that it is helpful for the integrated management of X. affinis.

Ultrastructure Characteristics and Sexual Dimorphism of Antennal Sensilla in Tirathaba rufivena (Lepidoptera: Pyralidae).

Tirathaba rufivena Walker, a major insect pest of Areca catechu L., has severely threatened areca nut cultivation in Hainan, China. To improve our understanding of the communication mechanism in host plant seeking and mate-finding for T. rufivena, we described and further characterized the external morphology and internal sensilla structures using scanning electron microscopy and transmission electron microscopy in this study. The antennal morphology was similar between males and females, and there was no significant difference in length between the two sexes. In total, nine sensilla types were identified: sensilla trichodea (Str), sensilla chaetica (Sch), sensilla basiconica (Sba), sensilla auricillica (Sau), sensilla coeloconica (Sco), sensilla styloconica (Sst), Böhm sensilla (Bs), uniporous peg sensilla (Ups) and sensilla squamiformia (Ssq). Sexual dimorphism mainly occurs in variation in the length of Sba, Sch, Sco1 and Bs, and the abundance of Sba, Sau1 and Sau2. The Sba had larger size and numbers on female antennae than that on males, suggesting that these sensilla might have important roles in locating host plants. Both Sau1 and Sau2 were significantly more abundant in females and were probably associated with the detection of mates and host plant for oviposition. These data were important for ongoing studies on host plant seeking and mate-finding behavior in T. rufivena and provided a theoretical foundation to further studies of semiochemical control for this pest.

Binding Properties of Odorant-Binding Protein 4 of Tirathaba rufivena to Areca catechu Volatiles.

Odorant-binding proteins (OBPs) play a key role in the olfactory system and are essential for mating and oviposition host selection. Tirathaba rufivena, a serious lepidopterous insect pest of the palm area in recent years, has threatened cultivations of Areca catechu in Hainan. Female-biased odorant-binding protein 4 of T. rufivena (TrufOBP4) expression was hypothesized to participate in the process of oviposition host recognition and localization. In this study, we cloned and analyzed the cDNA sequence of TrufOBP4. The predicted mature protein TrufOBP4 is a small, soluble, secretory protein and belongs to a classic OBP subfamily. Fluorescence binding assay results showed that TrufOBP4 had high binding abilities with the host plant volatiles, octyl methoxycinnamate, dibutyl phthalate, myristic acid and palmitic acid. These four components tend to dock in the same binding pocket based on the molecular docking result. The interactions and contributions of key amino acid residues were also characterized. This research provides evidence that TrufOBP4 might participate in the chemoreception of volatile compounds from inflorescences of A. catechu and can contribute to the integrated management of T. rufivena.

Antennal transcriptome analysis and candidate olfactory genes in Crematogaster rogenhoferi.

Crematogaster rogenhoferi (Hymenoptera: Formicidae), an omnivorous ant, is one of the dominant predatory natural enemies of a soft scale pest, Parasaissetia nigra Nietner (Homoptera: Coccidae), and can effectively control P. nigra populations in rubber forests. Olfaction plays a vital role in the process of predation. However, the information about the molecular mechanism of olfaction-evoked behaviour in C. rogenhoferi is limited. In this study, we conducted antennal transcriptome analysis to identify candidate olfactory genes. We obtained 53,892 unigenes, 16,185 of which were annotated. Based on annotations, we identified 49 unigenes related to chemoreception, including four odourant-binding proteins, three chemosensory proteins, 37 odourant receptors, two odourant ionotropic receptors and three sensory neuron membrane proteins. This is the first report on the molecular basis of the chemosensory system of C. rogenhoferi. The findings provide a basis for elucidating the molecular mechanisms of the olfactory-related behaviours of C. rogenhoferi, which would facilitate a better application of C. rogenhoferi as a biological control agent.

A Novel Nanobody Directed against Ovine Myostatin to Enhance Muscle Growth in Mouse.

Myostatin (MSTN) is a member of the transforming growth factor beta superfamily and is a negative regulator of myogenesis. It has been shown to function by controlling the proliferation of myoblasts. MSTN inhibition is considered as a promising treatment for promoting animal growth in livestock. Nanobodies, a special antibody discovered in camel, have arisen as an alternative to conventional antibodies and have shown great potential when used as tools in different biotechnology fields, such as diagnostics and therapy. In this study, we examined the effect of MSTN inhibition by RMN on the muscle growth of mice. The results showed that RMN could specifically detect and bind MSTN, as well as inhibit MSTN activity. A significant increase in skeletal muscle mass was observed after intramuscular injection of RMN into mice. Enhanced muscle growth occurred because of myofiber hypertrophy. These results offer a promising approach to enhance muscle growth that warrants further investigation in domestic animals.

Molecular characterization and expression of six heat shock protein genes in relation to development and temperature in Trichogramma chilonis.

Trichogramma is a kind of egg parasitoid wasp that is widely used to control lepidopterous pests. Temperature is one of the main factors that determines the various life activities of this species, including development, reproduction and parasitism efficiency. Heat shock proteins (HSPs) are highly conserved and ubiquitous proteins that are best known for their responsiveness to temperature and other stresses. To explore the potential role of HSPs in Trichogramma species, we obtained the full-length cDNAs of six HSP genes (Tchsp10, Tchsp21.6, Tchsp60, Tchsp70, Tchsc70-3, and Tchsp90) from T. chilonis and analyzed their expression patterns during development and exposure to temperature stress. The deduced amino acid sequences of these HSP genes contained the typical signatures of their corresponding protein family and showed high homology to their counterparts in other species. The expression levels of Tchsp10, Tchsp21.6 and Tchsp60 decreased during development. However, the expression of Tchsc70-3 increased from the pupal stage to the adult stage. Tchsp70 and Tchsp90 exhibited the highest expression levels in the adult stage. The expression of six Tchsps was dramatically upregulated after 1 h of exposure to 32 and 40°C but did not significantly change after 1 h of exposure to 10 and 17°C. This result indicated that heat stress, rather than cold stress, induced the expression of HSP genes. Furthermore, the expression of these genes was time dependent, and the expression of each gene reached its peak after 1 h of heat exposure (40°C). Tchsp10 and Tchsp70 exhibited a low-intensity cold response after 4 and 8 h of exposure to 10°C, respectively, but the other genes did not respond to cold at any time points. These results suggested that HSPs may play different roles in the development of this organism and in its response to temperature stress.

Molecular Characterization and Expression of Vitellogenin and Vitellogenin Receptor of Thitarodes pui (Lepidoptera: Hepialidae), an Insect on the Tibetan Plateau.

Vitellogenin (Vg) and vitellogenin receptor (VgR) play important roles in the vitellogenesis of insects. In this study, we cloned and characterized the two corresponding genes (TpVg and TpVgR) in an economically important insect, Thitarodes pui (Lepidoptera: Hepialidae), from the Tibetan plateau. The full length of TpVg is 5566 bp with a 5373 bp open reading frame (ORF) encoding 1,790 amino acids. Sequence alignment revealed that TpVg has three conserved domains: a Vitellogenin_N domain, a DUF1943 domain, and a von Willebrand factor type D domain (VWD). The full length of TpVgR is 5732 bp, with a 5397 bp ORF encoding 1798 amino acids. BLASTP showed that TpVgR belongs to the low-density lipoprotein receptor (LDLR) gene superfamily. Structural analysis revealed that TpVgR has a group of four structural domains: a ligand-binding domain (LBD), an epidermal growth factor (EGF)-precursor homology domain, a transmembrane (TM) domain, and a cytoplasmic domain. In addition, TpVgR has four cysteine-rich LDL repeats in the first ligand-binding site and seven in the second. Quantitative real-time polymerase chain reaction analysis revealed that the expression levels of TpVg and TpVgR are much higher in later pupa than in either the larval or adult stage, implying that the synthesis and uptake of Vg in T. pui occurs in the later pupal stage. These results will help us to understand the molecular mechanism of the reproductive capacity and will provide new insight into the mass rearing and utilization of T. pui.

A preliminary investigation of noise impact within metro stations in the urban city of Guangzhou, China.

The noise in the metro station is synthesized from a variety of different noise sources. Excessive noise exposure will bring serious impacts on humans' health. To alleviate the shortage of the urban land, most metro stations are planning to develop convenience stores and shopping malls. In order to evaluate the potential noise impact on humans' comfort in the metro stations, this study selected four representative stations of Guangzhou Metro Line 1 to carry out a preliminary questionnaire survey of 226 respondents for the perception and perform a noise measurement. Additionally, platform screen doors as a potential method for noise mitigation were examined. The results show that the noise caused by trains was dominant in the metro stations; however, the noise impact by public broadcast and passengers cannot be ignored. The noise levels of the transfer stations were obviously greater than the normal stations, especially during the peak hours. In addition, people in the metro stations have potential exposure to noise that the noise levels exceed the criteria limit of China, which would bring discomfort for humans; however, the normal activities of people would not be impacted. The platform screen doors should be further improved or relevant noise mitigation methods should be taken into account to reduce the noise level within the criteria limit.

Transcriptome characterization and gene expression analysis related to sexual dimorphism in the ghost moth, Thitarodes pui, a host of Ophiocordyceps sinensis.

Thitarodes pui is one of the host species of the Chinese caterpillar fungus Ophiocordyceps sinensis as a traditional Chinese medicine with economic and medical importance. The pupal and adult stages of T. pui are sexually dimorphic. In order to elucidate the molecular mechanisms involved in the sexually dimorphic development of T. pui, we compared the transcriptomes of female and male pupae and adults. We obtained 15,881,734, 16,962,086, 17,514,743, and 17,770,904 clean reads from female pupae, male pupae, female adults, and male adults, respectively. The reads obtained from the four samples were pooled and assembled into 65,165 unigenes, 23,597 of which were annotated. Candidate genes involved in sexual development were identified and analysed. Gene expression analysis revealed that 1406 genes were differentially expressed in male and female pupae, 448 of which were up-regulated in males and 958 were up-regulated in females. A total of 2025 genes were differentially expressed in male and females adults, 1304 of which were up-regulated in males and 721 were up-regulated in females. The functional enrichment of the differentially expressed genes indicated that reproduction and cuticle synthesis were regulated differently between the sexes. The transcriptome data obtained provide significant information regarding the genes involved in sexually dimorphic development, which will improve our understanding of the molecular mechanisms related to sexual dimorphism and helpful for the moth mass rearing which would provide enough host insects for the sustainable utilization of O. sinensis.

Effects of nickel exposure on testicular function, oxidative stress, and male reproductive dysfunction in Spodoptera litura Fabricius.

Nickel is an environmental pollutant that adversely affects the male reproductive system. In the present study, the effects of nickel exposure on Spodoptera litura Fabricius were investigated by feeding larvae artificial diets containing different doses of nickel for three generations. Damage to testes and effects on male reproduction were examined. The amount of nickel that accumulated in the testes of newly emerged males increased as the nickel dose in the diet increased during a single generation. Nickel exposure increased the amount of thiobarbituric acid reactive substances and decreased the amount of glutathione in treatment groups compared with the control. The activity levels of the antioxidant response indices superoxide dismutases, catalase, and glutathione peroxidase in the testes showed variable dose-dependent relationships with nickel doses and duration of exposure. Nickel doses also disrupted the development of the testes by decreasing the weight and volume of testes and the number of eupyrene and apyrene sperm bundles in treatment groups compared with the control. When the nickel-treated males mated with normal females, fecundity was inhibited by the higher nickel doses in all three generations, but fecundity significantly increased during the second generation, which received 5 mg kg(-1) nickel. Hatching rates in all treatments significantly decreased in a dose-dependent manner in the three successive generations. The effects of nickel on these parameters correlated with the duration of nickel exposure. Results indicate assays of testes may be a novel and efficient means of evaluating the effects of heavy metals on phytophagous insects in an agricultural environment.

De novo transcriptome characterization of the ghost moth, Thitarodes pui, and elevation-based differences in the gene expression of its larvae.

Thitarodes pui larvae are the hosts of a medicinal fungus, Ophiocordyceps sinensis, and are naturally distributed at an altitude of 4100-4650 m on Segrila Mountain of the Tibetan Plateau. Here, we conducted transcriptome profiling of T. pui larvae based on the Illumina high-throughput sequencing platform. Subsequently, we explored elevation-based differences of T. pui by comparing gene expression profiles between larvae at high-altitude (natural conditions) and larvae exposed to short-term (2months) low-altitude conditions. A total of 105,935,208 clean reads were assembled into 70,048 unigenes with a mean length of 639 bp. All unigenes were searched against public databases, with 51.26% unigenes being successfully annotated in the NR, SWISS-PROT, EuKaryotic Orthologous Groups (KOG), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genome (KEGG) databases. A total of 11,846 unigenes were functionally classified into 239 KEGG pathways. Metabolism was the most represented pathway, with 4271 unigenes (36.05%). Using the transcriptome data as a reference, 629 (311 up-regulated/318 down-regulated) genes were differentially expressed by low-altitude larvae when compared with those of high-altitude larvae. The most significantly differentially expressed genes were annotated in the processes of carbohydrate metabolism, lipid metabolism, and respiration. This report provides valuable information about the T. pui transcriptome for future genomic studies, including how gene expression is altered in larvae reared at different elevations.

Measurement of ground and nearby building vibration and noise induced by trains in a metro depot.

Metro depots are where subway trains are parked and where maintenance is carried out. They usually occupy the largest ground areas in metro projects. Due to land utilization problems, Chinese cities have begun to develop over-track buildings above metro depots for people's life and work. The frequently moving trains, when going into and out of metro depots, can cause excessive vibration and noise to over-track buildings and adversely affect the living quality of the building occupants. Considering the current need of reliable experimental data for the construction of metro depots, field measurements of vibration and noise on the ground and inside a nearby 3-story building subjected to moving subway trains were conducted in a metro depot at Guangzhou, China. The amplitudes and frequency contents of velocity levels were quantified and compared. The composite A-weighted equivalent sound levels and maximum sound levels were captured. The predicted models for vibration and noise of metro depot were proposed based on existing models and verified. It was found that the vertical vibrations were significantly greater than the horizontal vibrations on the ground and inside the building near the testing line. While at the throat area, the horizontal vibrations near the curved track were remarkably greater than the vertical vibrations. The attenuation of the vibrations with frequencies above 50 Hz was larger than the ones below 50 Hz, and the frequencies of vibration transmitting to adjacent buildings were mainly within 10-50 Hz. The largest equivalent sound level generated in the throat area was smaller than the testing line one, but the instantaneous maximum sound level induced by wheels squeal, contact between wheels and rail joints as well as turnout was close to or even greater than the testing line one. The predicted models gave a first estimation for design and assessment of newly built metro depots.