OBJECTIVETo study the chemical constituents of Bombyx batryticatus.

METHODThe chemical constituents were isolated from the title materials by column chromatography using silica gel, purified by crystallization, and identified by spectroscopic methods.

RESULTSeven compounds were isolated and identified.

CONCLUSIONSix compounds were isolated from Bombyx batryticatus for the first time.

Animals ; Bombyx ; chemistry ; Mannitol ; chemistry ; isolation & purification ; Materia Medica ; chemistry ; Palmitic Acid ; chemistry ; isolation & purification ; Sitosterols ; chemistry ; isolation & purification

OBJECTIVETo analysis the nutrient and effective ingredients of in Cordyceps militaris and make the best use of its medical value.

METHODAdenosine, cordycepin, polysaccharides, cordyceps acid, protein and fat in different parts of C. militaris were extracted, they are quantified by HPLC and other colorimetric analysis.

RESULTThe contents of polysaccharide was found to be 86.49 mg x g(-1) in C. militaris, 6.82 mg x g(-1) of adenosine in stroma, 13.28 mg x g(-1) of cordycepin and 44.07 mg x g(-1) of cordyceps acid in sclerolium.

CONCLUSIONIn different parts of C. militaris, the biosynthesis of effective ingredients is different. The total amount of effective ingredients is highest in C. militaris, the production of cordycepin and cordyceps acid is highest in sclerotium in comparison with other parts. Growth of C. militaris largely relies on its capability to utilize fat and protein from silkworm.

Adenosine ; analysis ; Animals ; Bombyx ; chemistry ; microbiology ; Cordyceps ; chemistry ; Deoxyadenosines ; analysis ; Fungal Proteins ; analysis ; Polysaccharides ; analysis

OBJECTIVEThe approach of this paper is make extracting solution' clarity achieve the request level and reduce the pollution of resinic columns through improved technical procedure. Silkworm dropping have sediment with alcohol extract and acid precipitation, affect the clarify performance quality, and polysaccharide and protein in extracting solution is badly for resinic columns in purification process.

METHODAdded freeze-thaw pretreatment in clarified process and researched three thaw methods effect on clarify performance of extracting solution with silkworm dropping. The adding effects were evaluated by content of polysaccharide and protein, transmittance and the yield of active ingredients during this adding process.

RESULTThrough comparing researches of three thaw methods, the microwave thaw method was the best approach.

CONCLUSIONAdding freeze-thaw pretreatment in clarify process is not only can improve the clarity of extracting solution with silkworm dropping on ideal level, reduce pollution of resinic columns, but also remain ideally the active ingredients.

Animals ; Biological Factors ; isolation & purification ; Bombyx ; chemistry ; Chemical Fractionation ; instrumentation ; methods ; Feces ; chemistry ; Freezing

The observation statistics suggested that the haemolymph melanization speed of larvae became fast and the growth inhibition of Escherichia coli was strong as the quantities of feeding on mulberry leaves increased. The RT-PCR result showed that the mRNA expressions of melanin biosynthesis enzyme BmTan, BmPo-1, BmYellow-f and BmDdc were high in the haemolyph of 5 L 3 d larvae. The qPCR analysis showed Bmtan, Bmddc, Bmyellow, Bmebony and Bmblack, especially Bmddc expression were significantly higher in black disease larvae than in normal larvae. Compared with control, Ddc inhibitors drastically inhibited the lipopolysaccharide-induced haemolymph melanization. In addition, the content of Dopa and Dopamine markedly rose after E. coli injection. These indicated that haemolymph melanization was linked to immune defenses and Bmddc may play a role in melanization response of haemolymph immune in silkworm.
Animals ; Bombyx ; enzymology ; genetics ; microbiology ; Escherichia coli ; Genes, Insect ; Hemolymph ; chemistry ; Larva ; Melanins ; biosynthesis

In silkworm moth the colleterial gland markedly enlarged due to the secretion and accumulation of glue like substances before adult emergence. However, the Ng mutant female moth only secreted little glue-like substance and laid loose eggs naturally. In the present experiment, it was extracted the proteins of secretory part of the variety E981 and its Ng mutant line and analyzed by two-dimensional electrophoresis. More than 700 protein spots were resolved both in two samples and most of the proteins were distributed in the area from 30 kD to 70 kD and pH 4 - 8. Through the comparison and analysis, it was found that 4 proteins were only expressed in E981 and 2 proteins were only expressed in Ng mutant. Furthermore, there are about 29 proteins were expressed higher in 981 and about 15 proteins expressed volume were higher in Ng mutant. These differential proteins may be have some relations with the Ng mutant form and directly lead to the Ng mutant can't secret the glue-like substance.
Animals ; Bombyx ; metabolism ; Electrophoresis, Gel, Two-Dimensional ; Exocrine Glands ; chemistry ; Female ; Insect Proteins ; analysis

Compared with herbal drugs, medicine processed from animals (animal medicine) was thought to have more bioactive substances and higher activities. Biotransformation effect often plays an important role in their effect. However, researches about effect of animal medicine on diabetic nephropathy and applying animal medicine as natural bio-transformer were seldom reported. The purpose of this paper was to reveal the use of Bombyx Mori L. on diabetic nephropathy from ancient to modern times. The classical literature indicated that Saosi Decoction (), which contains Bombyx Mori L. or silkworm cocoon, was applied to treat disorders congruent with modern disease diabetic nephropathy from the Ming to Qing Dynasty in ancient China. Modern studies showed that Bombyx Mori L. contains four main active constituents. Among these, 1-deoxynojirimycin (1-DNJ) and quercetin showed promising potential to be new agents in diabetic nephropathy treatment. The concentrations of 1-DNJ and the activities of quercetin in Bombyx Mori L. are higher than in mulberry leaves, because of the biotransformation in the Bombyx Mori L. body. However, these specifific components need further human and mechanistic studies to determine their therapeutic potential for this challenging condition.
Animals ; Biological Products ; therapeutic use ; Biotransformation ; Bombyx ; chemistry ; Diabetic Nephropathies ; drug therapy ; Medicine, Chinese Traditional

The aim of this study was to elucidate the scientific connotation of Bombyx Batryticatus processing with wheat bran under high temperature. The contents of soluble protein extracted from Bombyx Batryticatus and its processed products and the limited content of AFT in Bombyx Batryticatus and the processed one were compared. The concentration of protein was measured with the Bradford methods and the difference of protein between Bombyx Batryticatus and its processed products was compared by SDS-PAGE analysis. Aflatoxin B1, B2, G1, and G2 were determined by reversed-phase HPLC. The results showed that the soluble protein content of Bombyx Batryticatus and its processed products were (47.065 +/- 0.249), (29.756 +/- 1.961) mg x g(-1), correspondingly. Analysis of protein gel electrophoresis showed that there were no significant differences between the crude and processed one in protein varieties. 6 bands were detected: 31.90, 26.80, 18.71, 15.00, 10.18, 8.929 kDa. Below 10 kDa, the color of bands of the processed one was deeper than the crude one, which demonstrate that macromolecular protein was degradated into micromolecule. The content of AFG1, AFB1, AFG2, AFB2 were 0.382, 0.207, 0.223, 0.073 g x kg(-1), not exceeded 5 microg x kg(-1) while the processed one was not detected. Through processing with wheat bran under high temperature, the content of soluble protein in Bombyx Batryticatus decreased, the processing purpose for alleviating drug property was achieved. Meanwhile, the limited content of aflatoxins were reduced or cleared by processing procedure or absorbed by processing auxillary material, adding the safety of the traditional Chinese Medicine. In conclusion, as a traditional processing method, bran frying Bombyx Batryticatus was scientific and reasonable.
Animals ; Bombyx ; chemistry ; Chromatography, High Pressure Liquid ; Electrophoresis, Polyacrylamide Gel ; Hot Temperature ; Insect Proteins ; chemistry ; Molecular Weight

Starting with the relationship between mulberry leaves and silkworm droppings as food and metabolites, this study systematically compared the chemical components, screened out differential components, and quantitatively analyzed the main differential components based on ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) and UPLC-Q-TRAP-MS combined with principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). Moreover, the in vitro enzymatic transformation of the representative differential components was studied. The results showed that(1) 95 components were identified from mulberry leaves and silkworm droppings, among which 27 components only exist in mulberry leaves and 8 components in silkworm droppings. The main differential components were flavonoid glycosides and chlorogenic acids.(2) Nineteen components with significant difference were quantitatively analyzed, and the components with significant differences and high content were neochlorogenic acid, chlorogenic acid, and rutin.(3) The crude protease in the mid-gut of silkworm significantly metabolized neochlorogenic acid and chlorogenic acid, which may be an important reason for the efficacy change in mulberry leaves and silkworm droppings. This study lays a scientific foundation for the development, utilization, and quality control of mulberry leaves and silkworm droppings. It provides references for clarifying the possible material basis and mechanism of the pungent-cool and dispersing nature of mulberry leaves transforming into the pungent-warm and dampness-resolving nature of silkworm droppings, and offers a new idea for the study of nature-effect transformation mechanism of traditional Chinese medicine.
Animals ; Bombyx ; Morus/chemistry* ; Chlorogenic Acid/analysis* ; Gas Chromatography-Mass Spectrometry ; Chromatography, High Pressure Liquid/methods* ; Plant Leaves/chemistry*

The aim of this study was to prepare tandem multimeric proteins of BmSPI38, a silkworm protease inhibitor, with better structural homogeneity, higher activity and stronger antifungal ability by protein engineering. The tandem multimeric proteins of BmSPI38 were prepared by prokaryotic expression technology. The effects of tandem multimerization on the structural homogeneity, inhibitory activity and antifungal ability of BmSPI38 were explored by in-gel activity staining of protease inhibitor, protease inhibition assays and fungal growth inhibition experiments. Activity staining showed that the tandem expression based on the peptide flexible linker greatly improved the structural homogeneity of BmSPI38 protein. Protease inhibition experiments showed that the tandem trimerization and tetramerization based on the linker improved the inhibitory ability of BmSPI38 to microbial proteases. Conidial germination assays showed that His₆-SPI38L-tetramer had stronger inhibition on conidial germination of Beauveria bassiana than that of His₆-SPI38-monomer. Fungal growth inhibition assay showed that the inhibitory ability of BmSPI38 against Saccharomyces cerevisiae and Candida albicans could be enhanced by tandem multimerization. The present study successfully achieved the heterologous active expression of the silkworm protease inhibitor BmSPI38 in Escherichia coli, and confirmed that the structural homogeneity and antifungal ability of BmSPI38 could be enhanced by tandem multimerization. This study provides important theoretical basis and new strategies for cultivating antifungal transgenic silkworm. Moreover, it may promote the exogenous production of BmSPI38 and its application in the medical field.
Animals ; Antifungal Agents/pharmacology* ; Escherichia coli/metabolism* ; Proteins/metabolism* ; Protease Inhibitors/chemistry* ; Bombyx/chemistry* ; Saccharomyces cerevisiae/metabolism* ; Peptide Hydrolases

Two kinds of gel materials were prepared from silk fibroin with polyepoxy compound at subfreezing temperature and higher temperature. In order to evaluate the feasibility of their application in biomaterials, we tested the cytotoxicity of silk fibroin gels by detecting the effect of the extracted liquid on the cell relative proliferation rate of L-929 mouse fibroblasts. The results indicated that both of the gel materials displayed high relative proliferation rate and grade 1 cytotoxicity, being in the allowed range of medical application. The cytotoxicity tests on polyepoxy compound and glutaraldehyde were conducted too, and the cytotoxicity of polyepoxy compound was obviously lower than that of glutaraldehyde. Polyepoxy compound can be used as a more safe cross-link reagent for silk fibroin modification.
Animals ; Biocompatible Materials ; chemistry ; Bombyx ; chemistry ; Cross-Linking Reagents ; chemistry ; Fibroblasts ; drug effects ; Fibroins ; chemistry ; Gels ; chemical synthesis ; toxicity ; Mice ; Polyethylene Glycols ; chemistry ; Temperature