JRMS J Res Med Sci Journal of Research in Medical Sciences Journal of Research in Medical Sciences 1735-1995 1735-7136 Medknow Publications Pvt Ltd India JRMS-18-534 24250709 Letter to Editor Analysis of serum peptidome patterns of patients stung by honey bees Sui Weiguo Li Fengyan Hou Yuwen Dai Yong Nephrology Department of the 181st Hospital, Guangxi Key Laboratory of Metabolic Diseases Research, Central Laboratory, Guilin, China Nephrology Department of the 181st Hospital, Guangxi Key Laboratory of Metabolic Diseases Research, Central Laboratory, Guilin, China Nephrology Department of the 181st Hospital, Guangxi Key Laboratory of Metabolic Diseases Research, Central Laboratory, Guilin, China Nephrology Department of the 181st Hospital, Guangxi Key Laboratory of Metabolic Diseases Research, Central Laboratory, Guilin, China Address for correspondence:Yong Dai, Clinical Medical Research Center,The Second Clinical Medical College of Jinan University, Shenzhen People′s Hospital,Shenzhen,Guangdong, 518020, China daiyong2222@gmail.com June 2013 18 6 534 535 Copyright: © Journal of Research in Medical Sciences 2013

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<p>Sir,</p> <p>Recently, the number of the people who died from bitten by honeybees is gradually increasing. Hymenoptera venom of component is extremely complicated, and can impact function of multi-organ in vivo, such as intravascular hemolysis, rhabdomyolysis, acute renal failure, liver loss, heart loss, nervous system loss, disseminated intravascular coagulation, etc., Especially, the renal damage is common. <sup> <xref ref-type="bibr" rid="ref1">1</xref> </sup>Stings of Hymenoptera may lead to anaphylaxis. Anaphylaxis to insect stings has occurred in 3% of adults and can be fatal even on the first reaction. The body systems of patients stung by honey bees exert differentially expression proteins, which can threat human lives. <sup> <xref ref-type="bibr" rid="ref2">2</xref> </sup>2-D gel electrophoresis (2-DE) combined with protein identification by mass spectrometry (MS) is the most classic technology used for proteomics and is now recognized as an accurate method to determine and quantify human proteins, can reduce inter-gel variability and simplifying gel analysis. It is also a rapid and sensitive method. <sup> <xref ref-type="bibr" rid="ref3">3</xref> </sup></p> <p>We used the technique of 2-DE to separate differentially expression proteins in the serum of patients <xref ref-type="fig" rid="F1">Figure 1</xref>stung by honey bees, and compared with healthy control individuals <xref ref-type="fig" rid="F2">Figure 2</xref>. Then, we identified the bee venom-associated key protein molecules by MS analysis. The function and classification of these proteins were determined by searching the public database. <fig id="F1"> <label>Figure 1</label> <caption> <p>Patient group gel. The pH direction of the IEF is indicated on the top of the figure. Isoelectric Focusing pH 4~9</p> </caption> <alt-text>Figure 1</alt-text> <graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="JResMedSci_2013_18_6_534_119422_u1.tif" /> </fig> <fig id="F2"> <label>Figure 2</label> <caption> <p>Control group gel. The pH direction of the IEF is indicated on the top of the figure. Isoelectric Focusing pH 4~9</p> </caption> <alt-text>Figure 2</alt-text> <graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="JResMedSci_2013_18_6_534_119422_u2.tif" /> </fig></p> <p>Protein samples were separated on 13 cm linear pH 4-9 IPG strips, followed by sodium salt-Polyacrylamide gel electrophoresis (SDS-PAGE), and stained with Coomassie Blue R-250. The images were analyzed using the image master 2D platinum 5.0 software. An effective exclusion, in advance, of the given high-abundance proteins is the key to a successful 2-DE. In this study, the trials consequently contributed to the methods in the exclusion of serum albumin and Immunoglobulin G (IgG) through affinity and absorption by ProteoSeek <sup>TM</sup>Albumin/IgG Removal Kit, the purification of the protein samples by 2D clean-up kit and also an improved protocol of silver staining. <sup> <xref ref-type="bibr" rid="ref4">4</xref> </sup>This technique is very useful for applications requiring accurate quantization and direct differential proteomic analysis of some tissues or serum. <sup> <xref ref-type="bibr" rid="ref5">5</xref> </sup></p> <p>Another finding was that the abundances of some cytoskeletal proteins including Keratin1 type II (KRT1- II), alpha-2-macroglobulin and Spectrin beta chain, brain 1. They were decreased in the patients group. There has been reported alpha-2-macroglobulin which related to patients with bees venom. However, there is still one protein, protein-coding gene SDA1, domain containing (Sdad1), whose function is totally undetermined. These novel candidates would be more worthy for further investigation.</p> <p>The goal of this preliminary work focuses on primary delineating comparative protein profiles of patients with sting by boney bees and wasps and healthy controls with two dimensional electrophoresis and matrix assisted laser desorption/ionization time of flight MS. The study shows the potential application of MALDI-TOF-TOF spectrometry proteomics in identifying protein changes and detecting interested biomarker candidates in some diseases.</p> <p>Acknowledgment</p> <p>The authors would gratefully acknowledge the financial support by funds received from Guangxi Natural Science Foundation (No. 2012GXNSFDA053017), Guangxi Key Laboratory of Metabolic Diseases Research (No. 11-031-33 and 12-071-32), and Guilin Scientific Research and Technology Development Program (No. 20110119-8-1). 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