Aims: This study was designed to investigate the antibacterial properties of the sericin protein Bombyx mori and Samia cynthia ricini against Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus strains. Methodology and results: Sericin protein was extracted from the cocoons of B. mori races A, B, C, D, F1-1, F1-2 and S. ricini from Indonesia using a degumming process. The extracted sericin protein was evaluated as anti-microbials using Kirby Breuer disc diffusion, spectrometer measurement in decreasing OD value and imaging scanning electron microscope (SEM). The tests revealed that sericin protein from B. mori was more capable of inhibiting bacteria S. aureus than E. coli. Sericin protein from S. ricini was also capable of strongly inhibiting both S. aureus and E. coli bacteria. The sericin proteins of B. mori and S. ricini were found to reduce the number of S. aureus and E. coli bacteria as the OD value decreased at a concentration of 9%. The SEM imaging suggests that sericin B. mori and S. ricini proteins caused changes in cell morphology in S. aureus and E. coli bacteria, resulting in bacterial cell function disruption and death. Conclusion, significance and impact of study Bombyx mori and S. ricini sericin proteins were found to act against S. aureus and E. coli bacteria. Therefore, sericin protein has a greater antibacterial activity against Gram-positive strain S. aureus.
Anti-Bacterial Agents ; Sericins ; Bombyx ; Lepidoptera ; Escherichia coli ; Staphylococcus aureus

OBJECTIVE: To explore the mechanism of Flos Puerariae and Semen Hoveniae in treatment of alcoholic liver injury (ALI) based on network pharmacology and molecular docking. METHODS: The information of chemical constituents and targets of Flos Puerariae and Semen Hoveniae was collected from TCMSP and Swiss databases, and the threshold values of oral bioavailability (OB) ≥ 30%, drug likeness (DL) ≥0.18 were used to screen the potential active compounds. The GeneCard and DrugBank databases were used to obtain the targets corresponding to ALI. The common targets were queried using Venn Diagram, and the network of PPI and Gene Ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed through DAVID and Reactome database. Autodock Vina software was used for molecular docking of potential ingredients and key targets. RESULTS: A total of 21 potential active compounds and 431 therapeutic targets were gathered in Flos Puerariae and Semen Hoveniae, which involved 273 biological functions, 90 KEGG pathways and 362 Reactome pathways. The GO functions involved protein binding, ATP binding, etc.; the KEGG pathways mainly included PI3K-Akt signaling pathway and TNF signaling pathway; the Reactome pathways contained signal transduction and immune system, etc. The results of molecular docking showed that 21 potential active ingredients had good affinity with the core targets Akt1, TP53 and IL-6. CONCLUSIONS The network pharmacology and molecular docking analysis demonstrate the synergetic effect of Flos Puerariae and Semen Hoveniae with multi-compounds, multi-targets and multi-pathways in the treatment of ALI; and also predict the possible medicinal substance, key targets and pathways, which provides clues for the new drug development and mechanism research.
Animals ; Computer Simulation ; Drugs, Chinese Herbal/therapeutic use* ; Lepidoptera/chemistry* ; Liver/drug effects* ; Liver Diseases, Alcoholic/therapy* ; Molecular Docking Simulation ; Phosphatidylinositol 3-Kinases/metabolism* ; Plant Extracts/therapeutic use* ; Rhamnaceae/chemistry* ; Signal Transduction/drug effects*

Heortia vitessoides is the most serious pest of Aquilaria sinensis,which is an economically important evergreen tree native to China and is the principal source of Chinese agarwood. In severe infestations,the insects completely eat up the leaves of A. sinensis,causing severe economic losses. In a more recent study,we found that the antennal sensilla of adult play important roles in the host location,mating and oviposition of H. vitessoides. Here,the external morphology of the antennal sensilla of H. vitessoides were examined using scanning electron microscopy. The result showed that the antennae of both sexes of H. vitessoides were filiform in shape,which consist of the scape,pedicel and about 64 segments of flagellomeres. Eight morphological sensilla types were recorded in both sexes,including sensilla trichodea,sensilla chaetica,sensilla basiconica,sensilla coeloconica,sensilla styloconica,sensilla auricillica,sensilla squamiformia and böhm bristle. Major differences were recorded in the distribution and quantity of different sensilla types in each segment of antenna. The sensillas are almost confined to the ventral and lateral surfaces rather than the back side of antennae. Antennal flagella contained the most sensilla while the scape and pedicel segments only contained böhm bristles and sensilla squamiformias. Sensilla trichodea Ⅲ were only found on male antennae. These results are discussed in relation to the possible roles of the sensilla types in the host location,mating and oviposition selection behavior of H. vitessoides.
Animals ; China ; Female ; Lepidoptera ; anatomy & histology ; Male ; Microscopy, Electron, Scanning ; Sensilla ; ultrastructure ; Thymelaeaceae ; anatomy & histology

Under indoors simulating natural growing condition, the 4th-instar Mythimna separata larvae were fed by using poi- son leaf disk method. The effect of total ginsenosides on the protective enzymes (PPO, T-SOD, CAT and POD) of M. separata larvae was studied. The total ginsenosides could influence the protective enzymes of 4th-instar M. separata larvae significantly. After treated by total ginsenosides, the PPO activities increased firstly then decreased, and tended to equilibrium, and reached the maximum after 48 h. Furthermore, the total ginsenosides disturbed the dynamic balance of SOD, CAT and POD of M. separata larvae, and the yield of O2-* speeded. The results suggest that the total ginsenosides influence the protective enzymes of 4th-instar M. separata larvae, and disturb the original dynamic balance of protective enzymes. Consequently the insect suffers from the harm of O2-*.
Animals ; Enzymes ; metabolism ; Ginsenosides ; metabolism ; Larva ; metabolism ; Lepidoptera ; metabolism ; Oxygen ; metabolism

To address the role of Pr1 gene in the process of Hirsutella sinensis infecting Hepialus gonggaensis, differential expression of subtilisin-like protease gene was detected. In the present study, Pr1 gene analogues from H. sinensis were obtained by PCR strategy using specific primers designed from conserved regions of Pr1 gene reported in the GenBank. Then we detected the changes in the expression of Pr1 gene before and after infecting H. gonggaensis using real-time quantitative PCR. We obtained the partial sequence of Pr1 gene with the length of 535 bp (GenBank accession: KC009680). Real-time PCR results showed that the expression level of Pr1 gene was significantly different among 8 samples (P < 0.01). Pr1 gene showed the obvious higher expression level (2-3 folds) after infecting the H. gonggaensis, suggesting that the Pr1 gene may play an important role in the process of H. sinensis infecting H. gonggaensis. The present study paves a way for further identification on infectivity assessment of H. sinensis.
Amino Acid Sequence ; Animals ; Hypocreales ; genetics ; metabolism ; pathogenicity ; Larva ; Lepidoptera ; microbiology ; Real-Time Polymerase Chain Reaction ; Subtilisin ; genetics ; metabolism

Chinese Caterpillar Fungus (CCF) is one of the rare Chinese traditional drugs. As the resource is reducing sharply, the price is rising higher and higher, and there have been much more adulterants in the markets, but until now we don't have a scientific and accurate research on the identification study for this drug. On the basis of resource investigation, during the study of the samples collected by ourselves and the specimens stored in the museum, using the macroscopic and microscopic methods, referring to the literatures of entomology, emphasizing on the characteristics of polypide part, we have studied this species in detail of the macroscopic characters such as the insertion position of the stroma part, the annulations and segments of the caterpillar, the abdominal leg, the pinaculum, and the microscopic characters of the body wall; firstly added the microscopic character of the crotchets on the planta of abdominal leg. The result turned out that the characters which we have studied are regular and stable, and it have laid the foundation for the powder products and patent medicines which have used the crude drug of CCF.
Animals ; Cordyceps ; ultrastructure ; Larva ; anatomy & histology ; ultrastructure ; Lepidoptera ; anatomy & histology ; ultrastructure ; Materia Medica ; Medicine, Chinese Traditional

OBJECTIVETo evaluate the efficacy of Atalantia monophylla (A. monophylla) leaf in different solvent crude extracts and fractions against eggs of Spodoptera litura (S. litura).

METHODSHexane, ethyl acetate and chloroform solvent extracts of A. monophylla leaf and 12 fractions from hexane extract were screened at 5.0%, 2.5%, 1.0% and 0.5% for crude extracts and 1 000, 500, 250 and 125 mg/kg for fractions against the eggs of S. litura for the ovicidal activity. LC50 and LC90 were calculated using probit analysis.

RESULTSHexane crude extract showed maximum ovicidal activity of 61.94% at 5.0% concentration with a correlation value of r (2)=0.81, and least LC50 value of 3.06%. Hexane extract was fractionated using silica gel column chromatography and 12 fractions were obtained. Fraction 9 was active which showed maximum ovicidal activity of 75.61% at 1 000 mg/kg with the LC50 value of 318.65 mg/kg and LC90 value of 1 473.31 mg/kg. In linear regression analysis, significant and high correlation (r (2)=0.81%) was seen between concentration and ovicidal activity of hexane crude extracts and its active fraction.

CONCLUSIONSAs per our knowledge, this is the first report for ovicidal activity of A. monophylla against S. litura, A. monophylla could be used for the management of S. litura and other insect pests.

Animals ; Biological Assay ; Hexanes ; chemistry ; Humans ; Insecticides ; pharmacology ; Lepidoptera ; drug effects ; growth & development ; Plant Extracts ; pharmacology ; Plant Leaves ; chemistry ; Rutaceae ; chemistry ; physiology ; Spodoptera ; drug effects ; growth & development

To develop a method based on immunoreactions for detection of Ectropis obliqua Nucleopolyhedrovirus (EoNPV), the polyhedra of the virus were purified and used to immunize the mouse BALB/c. The spleen cells from the immunized mice were then fused with the myeloma cell line Sp2/0. A hybridoma cell line which can stably secrete the monoclonal antibody against EoNPV was achieved by using indirect ELISA screening and cloning methods, and was named as 7D3. Meanwhile, the polyhedrin gene was cloned from EoNPV and expressed in E. coli. Western blotting analysis showed that the monoclonal antibody prepared from 7D3 could specifically react with the recombinant polyhedrin. An indirect ELISA method based on this monoclonal antibody for detecting EoNPV in infected tea looper was developed.
Animals ; Antibodies, Monoclonal ; biosynthesis ; genetics ; immunology ; Antibody Specificity ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Hybridomas ; secretion ; Lepidoptera ; growth & development ; virology ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Viral Structural Proteins ; biosynthesis ; genetics ; immunology

cry1Ah1, one of holo-type cry genes, cloned in this laboratory from Bacillus thuringiensis strain has been patented in China, and it encoded a protein with strong insecticidal activity against certain lepidopteran insect pests, such as Chilo suppressalis. cry1Ah1 gene is exhibiting good application prospects. In order to improve the expression level of cry1Ah1 gene in rice, and investigate the effect of codon usage preference of gene expression, we designed five different optimized schemes for cry1Ah1 insecticidal critical fragment in accordance with bias of rice codon, to improve G+C content, removed the shear signal and unstable factors. Optimized cry1Ah1 genes were transformed into Escherichia coli Rosetta (DE3) respectively, and 65 kDa polypeptides was expressed normally in inclusion body separately. All of these expressed polypeptides showed insecticidal activity against 2nd-instar larvae of Plutella xylostella and neonate of Chilo suppressalis. After transformation with modified cry1Ah1 genes into Var nippobare, the transgenic rice seedlings were detected by PCR, the positive rate containing target gene was more than 87%. Afterwards, the results of real-time RT-PCR and ELISA assay indicated that the highest expression level of five modified cry1Ah1 genes was that using the highest frequent codons. Average expression amount of Cry1Ah1 polypeptides was 0.104% of total soluble proteins from the positive transgenic rice.
Animals ; Bacillus thuringiensis ; genetics ; metabolism ; Bacterial Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Codon ; genetics ; Endotoxins ; biosynthesis ; genetics ; Hemolysin Proteins ; biosynthesis ; genetics ; Insecticides ; Lepidoptera ; Oryza ; genetics ; Pest Control, Biological ; methods ; Plants, Genetically Modified ; genetics ; Recombinant Proteins ; biosynthesis ; genetics

OBJECTIVETo analyze the allergenicity and immunogenicity of Psilogramma menephron allergen so as to provide the basis for preparing recombinant and standardized allergen vaccines of Psilgramma menephorn.

METHODSThe extracts of Psilgramma menephorn were analyzed by SDS-PAGE, and the allergenicity and immunogenicity of the extracts were tested with 9 sera from allergic patients by means of immunoblotting.

RESULTSMore than 20 allergen proteins were separated from the extract of Psilgramma menephorn by SDS-PAGE, with the relative molecular weight ranging from 12,000 to 128,000. The relative molecular weight of the allergenic proteins were 74,000 (88.9%), 66,000 (22.2%), 49,000 (22.2%), 36,000 (77.8%), or 25,000 (33.3%), and those of the immunogenic proteins were 79,000 (33.3%), 74,000 (66.7%), 66,000 (22.2%), 49,000 (22.2%), 36,000 (44.4%), or 25,000 (55.6%).

CONCLUSIONThe relative molecular weight of the major allergenic proteins of Psilgramma menephorn are 74,000 and 36,000, and 74,000 and 25,000 for the major immunogenic proteins. These proteins constitute the major allergenic components for diagnosis and specific treatment of Psilgramma menephorn allergy.

Adolescent ; Adult ; Allergens ; immunology ; isolation & purification ; Animals ; Asthma ; blood ; immunology ; Blotting, Western ; Female ; Humans ; Immunoglobulin G ; blood ; Lepidoptera ; immunology ; Male ; Middle Aged ; Young Adult