Objective To study the growth regulation, environmental adaption and epigenetic regulation of Chrysomyia Megacephala pupae, in order to obtain the transcriptome data of Chrysomyia Megacephala in different growing periods, and lay the foundation for forensic application. Methods The Chrysomyia Megacephala was cultivated and after pupation, 3 pupae were collected every 24 h from pupation to emergence, and stored at -80 ℃ for later use. High-throughput sequencing was performed by Illumina Hiseq 4000 and Unigenes were obtained. The Unigenes were compared by comparison tool BLAST from NCBI in databases such as NR, STRING, SWISS-PROT （including Pfam）, GO, COG, KEGG in order to obtain the corresponding annotation information. The expression amount of Unigenes obtained by sequencing in Chrysomyia Megacephala in six different growing periods was calculated by FPKM method, and the discrepant genes were screened according to the following standards： the log₂ multiple absolute value of FPKM expression amount between two different growing periods must be larger than 1 （log₂|FC|>1）, and the false discovery rate must be less than 0.05. Results When the mean temperature was 25.6 ℃, Chrysomyia Megacephala emerged 6 d after they pupated. A total of 43 408 pieces of Unigenes were obtained and their mean length was 905 bp, of which 32 500, 18 720, 13 542, 9 191 and 18 720 pieces were annotated by NR, SWISS-PORT, Pfam, STRING and KEGG databases. According to the discrepant gene analysis of pupae in two different growing periods, the number of genes with variants ranged from 801 to 5 307, and the total number of discrepant genes was 45 676. Conclusion The gene expressions of the transcriptome data of Chrysomyia Megacephala pupae in different growing periods are different. The results provided a good foundation for further research on the transcriptome changes in each period of the pupae of sarcosaprophagous flies and provided the basis for exploring the genes associated with the growth of Chrysomyia Megacephala pupae.
Animals ; Epigenesis, Genetic ; Gene Expression Profiling ; High-Throughput Nucleotide Sequencing ; Molecular Sequence Annotation ; Pupa/genetics* ; Transcriptome

PURPOSE: To investigate the causative allergens and clinical characteristics of Korean adult patients with food allergy (FA). METHODS: This retrospective, cross-sectional single-institutional study enrolled Korean adult patients (n = 812) suspected of having FA. For diagnosis, causality assessment history taking, ImmunoCAP specific immunoglobulin E measurement and/or skin prick test were performed. RESULTS: Among 812 patients, 415 were diagnosed as having FA. The most common causative allergen was fruit, with a diagnosis of pollen food allergy syndrome (PFAS: 155, 37.3%), followed by crustaceans (111, 26.7%), wheat (63, 15.1%), fruits in patients without PFAS(43, 10.3%), buckwheat (31, 7.4%), peanut (31, 7.4%), walnut (25, 6.0%), red meat due to reaction to galactose-α-1,3-galactose (α-Gal) (8, 1.9%), and silkworm pupa (13, 3.1%). Allergy to egg, milk, fish, or shellfish was rare in Korean adults. One-third of patients with FA exhibited multiple FAs (238/415, 57.3%); the average number of causative allergens was 2.39. About 129 patients (31.0%) were diagnosed as having anaphylaxis; in these patients, wheat was the most frequent causative food. Twenty patients were further diagnosed with food-dependent exercise-induced anaphylaxis (FDEIA); all were due to wheat. In particular, crustaceans, wheat, PFAS, buckwheat, and red meat (α-Gal) were also frequent causes of anaphylaxis. CONCLUSIONS: Wheat, fruits with or without PFAS, and crustaceans are important and frequent causative allergens in Korean adult FA; these allergens differ from those found in childhood FA. It is notable that non-classic allergies, such as PFAS, FDEIA, and α-Gal allergy, are the important causes of anaphylaxis in Korean adult FA.
Adult ; Allergens ; Anaphylaxis ; Arachis ; Bombyx ; Diagnosis ; Fagopyrum ; Food Hypersensitivity ; Fruit ; Humans ; Hypersensitivity ; Immunoglobulin E ; Immunoglobulins ; Juglans ; Korea ; Milk ; Ovum ; Pollen ; Pupa ; Red Meat ; Retrospective Studies ; Shellfish ; Skin ; Triticum

OBJECTIVES: The Aedes mosquito is a vector for transmitting many arboviruses. Knowledge of the breeding habitat of this vector is vital for implementing appropriate interventions. Thus, this study was conducted to determine the breeding habitats and presence of Aedes mosquito species in the study areas.METHODS: A house-to-house cross-sectional survey of Aedes mosquito breeding habitats was carried out in Metema and Humera, Ethiopia, in August 2017. All available water-holding containers present in and around houses were inspected for the presence of immature stages of Aedes mosquitoes, and they were collected and reared to the adult stage for species identification. In the larval survey, the house index, container index, and Breteau index were computed as risk indices.RESULTS: Of the 384 houses surveyed for the presence of Aedes mosquito larval breeding, 98 were found to be positive for larvae. During the survey, a total of 566 containers were inspected, of which 186 were found to be infested with Aedes mosquito larvae, with a container index of 32.9, a house index of 25.5, and a Breteau index of 48.4. The most common Aedes mosquito breeding habitats were discarded tires (57.5%), followed by mud pots (30.0%). Of the 1,077 larvae and pupae collected and reared, Aedes aegypti (49.3%), Ae. vittatus (6.5%), and Culex species (44.2%) were identified.CONCLUSIONS: Discarded tires were the most preferred breeding habitats for Aedes mosquitoes. Moreover, Ae. aegypti, the main vector of dengue and other arboviruses, was identified for the first time in this region, suggesting a high potential for arbovirus transmission in the study areas.
Adult ; Aedes ; Arboviruses ; Breeding ; Cross-Sectional Studies ; Culex ; Culicidae ; Dengue ; Ecosystem ; Ethiopia ; Humans ; Larva ; Pupa

OBJECTIVES: The Aedes mosquito is a vector for transmitting many arboviruses. Knowledge of the breeding habitat of this vector is vital for implementing appropriate interventions. Thus, this study was conducted to determine the breeding habitats and presence of Aedes mosquito species in the study areas. METHODS: A house-to-house cross-sectional survey of Aedes mosquito breeding habitats was carried out in Metema and Humera, Ethiopia, in August 2017. All available water-holding containers present in and around houses were inspected for the presence of immature stages of Aedes mosquitoes, and they were collected and reared to the adult stage for species identification. In the larval survey, the house index, container index, and Breteau index were computed as risk indices. RESULTS: Of the 384 houses surveyed for the presence of Aedes mosquito larval breeding, 98 were found to be positive for larvae. During the survey, a total of 566 containers were inspected, of which 186 were found to be infested with Aedes mosquito larvae, with a container index of 32.9, a house index of 25.5, and a Breteau index of 48.4. The most common Aedes mosquito breeding habitats were discarded tires (57.5%), followed by mud pots (30.0%). Of the 1,077 larvae and pupae collected and reared, Aedes aegypti (49.3%), Ae. vittatus (6.5%), and Culex species (44.2%) were identified. CONCLUSIONS: Discarded tires were the most preferred breeding habitats for Aedes mosquitoes. Moreover, Ae. aegypti, the main vector of dengue and other arboviruses, was identified for the first time in this region, suggesting a high potential for arbovirus transmission in the study areas.
Adult ; Aedes* ; Arboviruses ; Breeding* ; Cross-Sectional Studies ; Culex ; Culicidae* ; Dengue ; Ecosystem* ; Ethiopia* ; Humans ; Larva ; Pupa

Mulberry (Morus alba) leaves are known to have therapeutic effects on lipid metabolism including lipogenesis, lipolysis and hyperlipidemia. However, novel compounds with strong lipolytic ability among 27 extracts of the mulberry leaves fermented with Cordyceps militaris (EMfCs) have not yet been identified. Therefore, the cAMP concentration and cell viability were measured in the primary adipocytes of SD (Sprague Dawley) rats and 3T3-L1 cells after treatment of 27 EMfCs. Briefly, mulberry leaves powders amended with three different concentrations (0, 25 and 50%) of silkworm pupae (SWP) powder were fermented with 10% C. militaris (v/w) during three different periods (3, 4 and 6 weeks). A total of 27 extracts were obtained from the fermented mulberry leaves powders using three different solvents (dH2O, 50% EtOH and 95% EtOH). Among the 27 EMfCs treated groups, a significant increase in the concentration of cAMP was detected in primary adipocytes treated with 10 extracts when compared with the Vehicle treated group. However, their cAMP concentration did not agree completely with the non-toxicity, although most extracts showed non-toxicity. Furthermore, the concentration of cAMP and level of free glycerol gradually increased in a dose dependent manner (100, 200 and 400 µg/mL) of 4M3-95 contained cordycepin without any significant toxicity. Overall, the results of this study provide strong evidence that 4M3-95 extract derived from EMfCs can stimulate the lipolysis of primary adipocytes at an appropriate concentration and therefore have the potential for use as lipolytic agents to treat obesity.
3T3-L1 Cells ; Adipocytes* ; Animals ; Bombyx ; Cell Survival ; Cordyceps* ; Glycerol ; Hyperlipidemias ; Lipid Metabolism ; Lipogenesis ; Lipolysis ; Morus* ; Obesity ; Powders ; Pupa ; Rats* ; Solvents ; Therapeutic Uses

Boiled silkworm pupa is a traditional food in Asia, and patients with silkworm pupa food allergy are common in these regions. Still now only one allergen from silkworm, arginine kinase, has been identified. The purpose of this study was to identify novel food allergens in silkworm pupa by analyzing a protein extract after heat treatment. Heat treated extracts were examined by proteomic analysis. A 27-kDa glycoprotein was identified, expressed in Escherichia coli, and purified. IgE reactivity of the recombinant protein was investigated by ELISA. High molecular weight proteins (above 100 kDa) elicited increased IgE binding after heat treatment compared to that before heat treatment. The molecular identities of these proteins, however, could not be determined. IgE reactivity toward a 27-kDa glycoprotein was also increased after heating the protein extract. The recombinant protein was recognized by IgE antibodies from allergic subjects (33.3%). Glycation or aggregation of protein by heating may create new IgE binding epitopes. Heat stable allergens are shown to be important in silkworm allergy. Sensitization to the 27-kDa glycoprotein from silkworm may contribute to elevation of IgE to silkworm.
Adolescent ; Adult ; Allergens/*chemistry/*immunology ; Amino Acid Sequence ; Animals ; Bombyx/*chemistry/genetics/growth & development/*immunology ; Epitopes/immunology ; Female ; Food Hypersensitivity/etiology ; Glycoproteins/*chemistry/genetics/*immunology ; Hot Temperature ; Humans ; Immunoglobulin E/immunology ; Male ; Molecular Sequence Data ; Molecular Weight ; Proteomics ; Pupa/chemistry/immunology ; Recombinant Proteins/biosynthesis/chemistry/immunology ; Sequence Alignment

Forensic entomology investigates postmortem interval (PMI) estimation using insect evidence. We estimated the minimum PMI of a putrefied male cadaver using pupae in the soil and maggots found in the body. Most of the maggots, collected during the autopsy, were postfeeding third instar larvae with empty crop contents, which developed freezing injuries when the cadaver was placed in the freezer. Pupae in the soil were collected 45 days after the discovery of the body. DNA barcoding revealed that most pupae and maggots were Chrysomya pinguis, with a few exceptions. The minimum PMI was estimated at 10 days before the discovery time based on the scene investigation, maggot developmental stage, distribution of pupae moving away from the body toward pupariation sites, DNA barcoding results, and weather information. To reduce the gap between the minimum and maximum PMI values, complete entomological evidence collection should be conducted at the time of discovery.
Autopsy ; Cadaver ; Diptera* ; DNA ; Entomology* ; Forensic Sciences ; Freezing* ; Humans ; Insects ; Larva* ; Male ; Postmortem Changes ; Pupa* ; Soil ; Weather

microRNAs (miRNAs) are an extensive class of -22-nucleotide (nt) endogenous noncoding RNAs regulating life activities ofmetazoans through binding to 3'-untranslated regions (3'-UTRs) of their target genes. This work aimed to identify yan gene in the silkworm, reveal its expression profile and confirm if it is one target of bmo-miR-7 and, as such, have potential for contributing to better understanding of the molecular mechanisms involved in the metamorphosis of silkworm. Based on homolog searching and PCR amplification, we cloned the coding sequence (CDS) of Bmyan, which encodes 476 amino acid residues and contains SAM-PNT and ETs domains. Quantitative PCR (q-PCR), RT-PCR and microarray data revealed high expression of Bmyan in the head, body wall and ovary of day-3 fifth instar larval silkworm, low or no expression in other tissues. It was lowly expressed in the early larval stages, but highly expressed from late spinning to day 4 pupa. The 3'-UTR of Bmyan was obtained by rapid-amplification of cDNA ends (3'RACE) and predicted to contain two potential recognition sites of bmo-miR-7. The luciferase reporter vector containing the 3'-UTR of Bmyan was constructed and co-transfected into BmE cell line with the mimic of bmo-miR-7 and the decreased relative activity of luciferase showed that Bmyan is one target of bmo-miR-7. This work helps further functional analysis of bmo-miR-7 and Bmyan in the silkworm.
3' Untranslated Regions ; Animals ; Bombyx ; genetics ; Cell Line ; Cloning, Molecular ; Female ; Genetic Vectors ; Insect Proteins ; genetics ; Larva ; Metamorphosis, Biological ; MicroRNAs ; genetics ; Pupa

OBJECTIVETo clone and identify olfactory receptor odorant receptor 7 (OR7) gene of Aedes albopictus and analyze its expression profile and calcium regulation function.

METHODRT-PCR was used to amplify the olfactory receptor OR7 gene of Ae. albopictus and OR7 expression was detected in different tissues and organs. The coding sequence of OR7 gene was cloned in eukaryotic expression vector pME18s, which was then transfected into HEK293 cells. The calcium callback function in response to odor molecule stimulation was analyzed by calcium imaging technique.

RESULTSThe OR7 gene of Ae. albopictus was cloned and sequence analysis showed that its coding region was 1395 bp. RT-PCR detected OR7 expression in the larvae, pupae and adult mosquitoes, especially in female mosquitos. Preliminary analysis of calcium callback function demonstrated the specific regulation of calcium absorption by OR7 in response to odor molecule stimulation.

CONCLUSIONThe OR7 gene of Ae. albopictus has been cloned successfully. OR7 is highly expressed in female mosquitos and is capable of specific recognition of the odor molecules.

Aedes ; genetics ; Animals ; Cloning, Molecular ; Female ; Gene Expression ; Genes, Insect ; HEK293 Cells ; Humans ; Larva ; Pupa ; Receptors, Odorant ; genetics

Cordyceps is a fungus used as a traditional medicine in China, Japan, and Korea. Paecilomyces (P.) japonica is a new cordyceps that was recently cultivated on silkworm pupae in Korea. The present study evaluated the toxicological effects of P. japonica in rats. Forty rats were treated with oral doses of P. japonica (0, 20, 100, or 500 mg/kg/day) for 4 weeks. Twenty additional rats were treated with 0 or 500 mg/kg/day of P. japonica for 4 weeks and then maintained for 2 weeks without treatment. Clinical signs, body weight, food and water consumption, and organ weight as well as hematology, serum biochemistry, and histopathology data were examined. Body weight gain of the group treated with 500 mg/kg/day was significantly reduced. Microscopically, karyomegaly, single cell necrosis, and mitosis were observed in the renal tubular epithelium of all treated groups. In conclusion, P. japonica caused a reduction of body weight and renal injury in rats. The no observed adverse effect level (NOAEL) of P. japonica was less than 20 mg/kg/day.
Animals ; Biochemistry ; Body Weight ; Bombyx ; China ; Cordyceps ; Drinking ; Epithelium ; Fungi ; Hematology ; Japan ; Korea ; Medicine, Traditional ; Mitosis ; Necrosis ; No-Observed-Adverse-Effect Level ; Organ Size ; Paecilomyces* ; Pupa ; Rats*