[Study on quality control of Holotrichia diomphalia larvae by TLC and HPLC].[Pubmed: 29676127]

The TLC method was established for identification of Holotricha diomphalia larvae and the HPLC method was used to determine the content of inosine and guanosine in H. diomphalia larvae. The HPLC analysis was performed on a Waters HSS T3（4.6 mm×250 mm, 5 μm） column of with mobile phase consisting of acetonitrile (A) and 0.08% trifluoroacetic acid (B) in gradient elution. The detection wavelength was 260 nm. The flow rate was 1.0 mL·min⁻¹. The column temperature was 30 °C. As a result, TLC identification method had a good reproducibility and highly specificity. The linear equations of inosine and guanosine were in good linear range (r>0.999 8). The average recovery of inosine and guanosine was 96.53% (RSD=1.6%), 99.71% (RSD=2.7%). The method is simple, accurate and reproducible, which can provide a basis for quality standard improvement H. diomphalia larvae.

Anticoagulant activity of crude extract of Holotrichia diomphalia larvae.[Pubmed: 26578186]

Holotrichia diomphalia larvae are one classical folk medicinal material in East Asia which has clinically been used to promote blood circulation and dispel blood stasis for several hundred years.

[Analysis of liposoluble constituents in Holotrichia diomphalia by GC-MS and investigation their anti-inflammatory and analgesic activities].[Pubmed: 22876670]

To analysis liposoluble constituents of Holotrichia diomphalia by GC-MS and measure their anti-inflammatory and analgesic activities.

Anatomical organization of antennal-lobe glomeruli in males and females of the scarab beetle Holotrichia diomphalia (Coleoptera: Melolonthidae).[Pubmed: 21889404]

The glomerular organization of the primary olfactory brain center, the antennal lobe, was studied in males and females of Holotrichia diomphalia adults using serial histological sections labeled by the reduced silver-stain technique. The results revealed an apparent sexual dimorphism. Whereas an enlarged cap-shaped glomerulus was found at the antennal nerve entrance into the antennal lobe in males, no such unit was present in females. Also the size of the antennal lobe differed between the sexes, the antennal lobe of males being larger than that of females. We estimated the total number of glomeruli at approximately 60 units in the female antennal lobe. In males, we could discriminate only those glomeruli that were located in the anterior area of the antennal lobe.

[Analysis of proteins, amino acids and inorganic elements in Holotrichia diomphalia from different areas].[Pubmed: 21355187]

To analyze the content of proteins,amino acids and inorganic elements of Holotrichia diomphalia in different growing areas as the references for quality evaluation and reasonable application of them.

Beetle immunity.[Pubmed: 21528698]

Genetic studies have elegantly characterized the innate immune response in Drosophila melanogaster. However, these studies have a limited ability to reveal the biochemical mechanisms underlying the innate immune response. To investigate the biochemical basis of how insects recognize invading microbes and how these recognition signals activate the innate immune response, it is necessary to use insects, from which larger amounts of hemolymph can be extracted. Using the larvae from two species of beetle, Tenebrio molitor and Holotrichia diomphalia, we elucidated the mechanisms underlying pathogenic microbe recognition. In addition, we studied the mechanism of host defense molecule amplification. In particular, we identified several pattern recognition proteins, serine proteases, serpins and antimicrobial peptides and examined how these molecules affect innate immunity.

Mechanisms of inhibiting proliferation of vascular smooth muscle cells by serum of rats treated with Dahuang Zhechong pill.[Pubmed: 19527826]

Dahuang Zhechong pill (DHZCP), a famous and classical Chinese herbal prescription, consists of twelve traditional Chinese drugs: Eupolyphaga sinensis Walker., Rheum officinale Baill., Scutellaria baicalensis Georgi., Glycyrrhiza uralensis Fisch., Prunus persica Batsch., Prunus armeniaca L., Paeonia lactiflora Pall., Rehmannia glutinosa Libosch., Toxicodendron vernicifluum F.A. Barkl., Tabanus bivittatus Mats., Hirudo nipponica Whitman. and Holotrichia diomphalia Bates., and is clinically used to treat hepatic diseases, gynecopathy and atherosclerosis in China. Our previous studies confirm that DHZCP is able to significantly inhibit proliferation of vascular smooth muscle cells (VSMCs) in vivo and in vitro.

Antifungal activity of crude extracts and fat-soluble constituents of Holotrichia diomphalia larvae.[Pubmed: 18424040]

In this study, chemical compositions of fatty oils and bioactivity of crude extracts from Holotrichia diomphalia larvae as Chinese materia medica were investigated for the first time. The chemical compositions of the fatty oils were obtained by two different methods and determined by GC/MS. In total, the petroleum ether extract produced 21 compounds (96.3%) while the supercritical fluid extract produced six compounds (99.53%) for identification. The effect of petroleum ether and other crude extracts on Pyricularia oryzae was also examined. Results indicated that ethanol and petroleum ether extracts had excellent antifungal activities. These findings demonstrated that fatty oils from H. diomphalia larvae had great potential to be used as a source for natural health products.

A novel 40-kDa protein containing six repeats of an epidermal growth factor-like domain functions as a pattern recognition protein for lipopolysaccharide.[Pubmed: 16849495]

Determination of structures and functions of pattern recognition proteins are important for understanding pathogen recognition mechanisms in host defense and for elucidating the activation mechanism of innate immune reactions. In this study, a novel 40-kDa protein, named LPS recognition protein (LRP), was purified to homogeneity from the cell-free plasma of larvae of the large beetle, Holotrichia diomphalia. LRP exhibited agglutinating activities on Escherichia coli, but not on Staphylococcus aureus and Candida albicans. This E. coli-agglutinating activity was preferentially inhibited by the rough-type LPS with a complete core oligosaccharide. LRP consists of 317 aa residues and six repeats of an epidermal growth factor-like domain. Recombinant LRP expressed in a baculovirus system also showed E. coli agglutination activity in vitro and was able to neutralize LPS by inhibition of LPS-induced IL-6 production in mouse bone marrow mast cells. Furthermore, E. coli coated with the purified LRP were more rapidly cleared in the Holotrichia larvae than only E. coli, indicating that this protein participates in the clearance of E. coli in vivo. The three amino-terminal epidermal growth factor-like domains of LRP, but not the three carboxyl epidermal growth factor-like domains, are involved in the LPS-binding activity. Taken together, this LRP functions as a pattern recognition protein for LPS and plays a role as an innate immune protein.

Overexpression and preliminary X-ray crystallographic analysis of prophenoloxidase activating factor II, a clip domain family of serine proteases.[Pubmed: 15953772]

A clip domain family of serine proteases has been identified in invertebrates as a crucial enzyme involved in diverse biological processes including immune responses and embryonic development. Although these proteins contain at least one clip domain at the N-terminal of the serine protease domain, the roles and three-dimensional structure of the clip domain are unknown. Prophenoloxidase activating factor-II (PPAF-II), a clip domain family of serine proteases, derived from the beetle Holotrichia diomphalia larvae, was overexpressed in the baculovirus system, and crystallized using the hanging-drop vapor-diffusion method. High-quality single crystals of PPAF-II were obtained in a precipitant solution containing 0.15 M ammonium sulfate, 1.25 M lithium sulfate monohydrate, and 0.1 M sodium citrate dehydrate (pH 5.5). These crystals belong to space group C2 with unit-cell parameters a=107.84, b=76.78, c=70.49 A and beta=113.93 degrees , and contain one or two molecules in the asymmetric unit. Determination of the three-dimensional structure of PPAF-II would clarify the functions of the clip domains.