1. Developed a temperature-controllable nano-liter electrospray ion source, which can effectively enhance the detection rate of hydrophobic peptides of membrane proteins.
Membrane protein analysis is a technological challenge in proteomics research due to its low abundance and hydrophobicity. Fractionation, extraction, digestion and subsequent low detection rate by mass spectrometry analysis should be considered in membrane protein analysis. The centrifugal proteomic reactor was developed for membrane protein sample preparation through the collaboration between Dr. Figeys' lab in University of Ottawa and us. To further improve the mass spectrometry detection capability of hydrophobic peptides from membrane proteins, we have developed a temperature-controlled nanoliter electrospray ionization source. The range of this ion source can control the column temperature from room temperature to 80 °C, and the temperature deviation is less than 0.3 °C. By heating the column temperature to 60 degrees, the detection rate of membrane proteins was effectively improved, especially the hydrophobic transmembrane peptides.

2. Established quantitative proteomics technology platform for drug action mechanism study.
We have established quantitative proteomics technology platform which can quantify the proteins changes from cell lines, primary cultured cells or tissues with or without drug treatments. Through collaboration with Prof. Zhang HY and Prof. Jiang HL' groups, quantitative proteomics was employed to reveal the neuroprotective effects of huperzine A towards amyloid beta treated neuroblastoma N2a cells. Huperzine A, a novel lycopodium alkaloid isolated from tranditional Chinese medicine Huperzia serrata (Qian Ceng Ta), has been shown have neuroprotective effects for Alzheimer's disease, but the action mechanism of huperzine A is unclear and need to be further investigated. In this study, proteins from untreated N2a cells (Con), cells pre-incubated with huperzine A followed by Aβ (1-42) oligomers treatment (HupA) and cells treated with Aβ (1-42) oligomers (Aβ) with 5 bilogcial replicates in each cohort, were processed in centrifugal proteomic reactor and quantified by label-free quantitation . A total of 2860 proteins were quantified with high confidence, and 198 proteins were significantly changed (with p-value <0.05) between HupA and Aβ cohorts. The pathway and direct protein-protein interaction network analysis showed that huperzine A protects N2a cells against Aβ oligomer-induced cell death by down-regulation of cellular tumor antigen p53 (Trp53) expression

3. Cooperate with other research group to promote the progress of related projects.
1) Cooperation with Prof. ZhangHY, we applied label-free quantitative proteomic approach to study iron damage to the primary cultured neurons. We found that proteins associated with formation of Aβ peptides were significantly changed. Subsequent western blot results showed that the changed proteins were consistent with mass-spectrometry based proteomics research. Further biological functional experiments are undergoing now.
2) Work together with Prof. Liu JG, quantitative proteomics study of the dorsal hippocampus from rat heroin self-administration model found that the formation of cytoskeletal protein and Wnt signaling pathway were involved in drug addiction. For example, β-catenin was found down-regulated after heroin treatment and, and the further experiments were performed for biological validation.
3) The transcription factor AbfR is involved in biofilm formation of Staphylococcus epidermidis. By collaboration with Prof. Yang CG’s group, quantitative proteomics is performed to compare the proteome from wild-type and Abfr-KO strains to investigate the mechanism of AbfR-mediated biofilm formation.

CAS-Bayer Young Scholar’s Award, CAS
Jul 30-Aug 2, 2009 The Young Scientist Program of the 21st IUBMB, Shanghai, China (116 candidates overall the world)

(2009.1.1- )
1. Ma Y, Yang C, Tao Y, Zhou H#, Wang Y#. Recent technological developments inproteomics shed new light on translational research on diabetic microangiopathy. FEBS J. 2013 Nov;280(22):5668-81. #co-corresponding authors

2. Tao YM, Fang L, Yang YM, Jiang HL, Yang HY# , Zhang HY# and Zhou H# Quantitative proteomic analysis reveals the neuroprotective effects of huperzine A for amyloid beta treated neuroblastoma N2a cells. Proteomics 2013 Apr;13(8):1314-24. #co-corresponding authors

3. Fowler SL, Akins M, Zhou H, Figeys D, Bennett SA. The liver connexin32 interactome is a novel plasma membrane-mitochondrial signaling nexus. J Proteome Res. 2013 Jun 7;12(6):2597-610.

4. Yao Z, Zhou H, Figeys D, Wang Y, Sundaram M. Microsome-associated lumenal lipid droplets in the regulation of lipoprotein secretion. Curr Opin Lipidol. 2013 Apr;24(2):160-70.

5. Wang F, Wei X, Zhou H, Liu J, Figeys D, Zou H. Combination of online enzyme digestion with stable isotope labeling for high-throughput quantitative proteome analysis. Proteomics. 2012 Nov;12(21):3129-37.

6. Zhou H, Ning Z, Starr AE, Abu-Farha M, Figeys D. Advancements in top-down proteomics. Anal Chem. 2012 Jan 17;84(2):720-34.

7. Zhou H, Ning Z, Wang F, Seebun D, Figeys D. Proteomic reactors and theirapplications in biology. FEBS J. 2011 Oct;278(20):3796-806.

8. Zhou H, Wang F, Wang Y, Ning Z, Hou W, Wright TG, Sundaram M, Zhong S, Yao Z, Figeys D. Improved recovery and identification of membrane proteins from rathepatic cells using a centrifugal proteomic reactor. Mol Cell Proteomics. 2011Oct;10(10):O111.008425.

9. Ning Z, Zhou H, Wang F, Abu-Farha M, Figeys D. Analytical aspects of proteomics: 2009-2010. Anal Chem. 2011 Jun 15;83(12):4407-26.