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Comparative Study on the Infectivity and Spore Surface Protein of Nosema bombycis and Its Morphological Variant Strain

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Agricultural Sciences in China 2005年第6期4卷 475-480页

作者： HUANGShao-kang LUXing-meng InvertebratePathologyLaboratory AnimalScienceCollegeZhejiangUniversityHangzhou310029P.R.China／／BeeScienceCollegeFujianAgricultureandForestryUniversityFuzhou350002P.R.China InvertebratePathologyLaboratory AnimalScienceCollegeZhejiangUniversityHangzhou310029P.R.China

A new morphological variant strain of microsporidium was produced by infecting the mulberry looper, Hemerophilaatrilineata [Phthonandria atrilineata], with Nosema bombycis successively for 24 times, and named 24Nbh. Comparativestudies on morphology, infectivity and spore surface protein were conducted. 24Nbh was short and wide, and had asignificant difference (P<0.01) over the Nb spores. The infectivity tests conducted on second instar silkworm larvaeshowed that IC50 of 24Nbh was 1.98104 spores mL-1 and of Nb was 1.72103 spores mL-1, thus indicating that the infectivityof Nb decreased 11.5 times after multiplying in mulberry looper for 24 times. The IC50 of spores from silkworm infected with24 Nbh was 6.9 times less than Nb, showing that the infectivity of 24Nbh spores rejuvenated very fast when reinfected tosilkworms, further more, the length and width of such spore was larger than 24Nbh (P<0.01) and smaller than Nb (P<0.05).The SDS-PAGE profiles of Nb and 24Nbh were generally the same, 4 distinct proteins of 12, 17, 30, 33 kDa were obtainedwith difference in quantity. When 120 g of protein was applied for 2D-PAGE, five suspected different proteins withdifference in quantity were observed. These results demonstrate that these differential proteins maybe associated withvariation in infectivity of the spores.

关键词： Nosema bombycis Morphological variation Infectivity Surface protein two dimensional electrophoresis

Identification of squamous cell carcinoma associated proteins by proteomics and loss of beta tropomyosin expression in esophageal cancer

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World Journal of Gastroenterology 2006年第44期12卷 7104-7112页

作者： Ferdous Rastgar Jazii Zahra Najafi Reza Malekzadeh Thomas P Conrads Abed Ali Ziaee Christian Abnet Mansour Yazdznbod Ali Asghar Karkhane Ghasem H Salekdeh National Institute of Genetic Engineering and Biotechnology POBOX: 14155 6343 Tehran Iran Digestive Disease Research Center Shariati Hospital Medical University Tehran Iran Laboratory of Proteomics and Analytical Technologies SAIC -Frederick B Frederick MD 21702 United States Institute of Biochemistry and Biophysics Tehran University Tehran Iran NCI Center for Cancer Research CancerPrevention Studies Branch Bethesda United States Department of surgery Shariati Hospital Medical University Tehran Iran Agricultural Biotechnology ResearchInstitute of Iran Karaj Iran

AIM： To assess the proteome of normal versus tumor tissue in squamous cell carcinoma of the esophagus （SCCE） in Iranian patients and compare our results with former reports by using proteomics. METHODS： Protein was extracted from normal and tumor tissues. two dimensional electrophoresis was carried out and spots with differential expression were identified with mass spectrometry. RNA extraction and RT-PCR along with immunodetection were performed. RESULTS： Fourteen proteins were found whose expression levels differed in tumor compared to normal tissues. Mass spectrometric analysis resulted in the identification of β-tropomyosin （TMβ）, myosin light chain 2 （and its isoform）, myosin regulatory light chain 2, peroxyredoxin 2, annexin I and an unknown polypeptide as the down regulated polypeptides in tumor tissue. Heat shock protein 70 （HSP70）, TPM4-ALK fusion oncoprotein 2, myosin light polypeptide 6, keratin I, GH16431p and calreticulin were the up-regulated polypeptides found in tumor tissue. Several of these proteins, such as TMβ, HSP70, annexin Ⅰ, calreticulin, TPM4-ALK and isoforms of myosins, have been well recognized in tumorigenesis of esophageal or other types of cancers. CONCLUSION： Our study not only supports the involvement of some of the formerly reported proteins in SCCE but also introduces additional proteins found to be lost in SCCE, including TMβ.

关键词： Squamous Cell Carcinoma Esophagus Esophageal Proteomics two dimensional electrophoresis Polypeptide marker

Difference of gene expression between the central and the peripheral epithelia of the bovine lens

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Chinese Medical Journal 2009年第9期122卷 1072-1080页

作者： MA Xuan WU Ming-xing ZHANG Yan-li CUI Dong-mei LI Ming-tao WU Kai-li Zhongshan Ophthalmic Center State Key Laboratory ofOphthalmology Sun Yat-sen University Guangzhou Guangdong510060 China Proteomics Laboratory Zhongshan Medical College Sun Yat-senUniversity Guangzhou Guangdong 510080 China Department of Ophthalmology Second People's Hospital ofYunnan Province Kunming Yunnan 650021 China

Background Equatorial lens epithelial cells proliferate and differentiate into fiber cells throughout life, while central lens epithelial cells proliferate little and do not form fiber ceils. This study aimed to investigate the differences in gene expression between the central and the peripheral epithelial cells of the bovine lens. Methods Lens epithelia were dissected into central （≤11.5 mm diameter, cLEC） and peripheral regions （pLEC）. The differences in gene expression and protein accumulation between these two regions were assayed by microarray analysis and two-dimensional electrophoresis （2-DE） combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS）. Differently expressed proteins were validated by immunoanalyses. Results By microarray analysis, 67 transcripts were at least two-fold lower and 269 at least two-fold higher in pLEC compared with that in cLEC. Thirty-four protein spots, including 20 in cLEC and 14 in pLEC, were identified by two dimensional electrophoresis and mass spectrometry. Of these 34 protein products, 28 were represented by probe sets on the microarray. Nine transcripts changed in the same direction and four transcripts in the opposite direction to their protein products. Immunoanalyses revealed that three （mitogen-activated protein kinase 1 （MAPK1）, nidogen （NID）, small nuclear ribonucleoprotein N （SNRPN）） out of four transcripts with opposite change between 2-DE and microarray assay showed the same changes as the results of 2-DE gel analyses. The genes differently expressed between cLEC and pLEC mainly include those related to the MAPK, transforming growth factor β （TGFβ） signaling and glycolysis pathways. Conclusion The results suggested that there were distinctly different genome activities, including a specific group of pathways, between central and peripheral lens epithelial cells.

关键词： microarray two dimensional electrophoresis mass spectrum lens epithelial cell

Secondary Structure and Neurotrophic Effect of a 33.1 kDa Specific Protein (SSP-33.1) in Spinal Sensory Ganglia

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Journal of Chinese Pharmaceutical Sciences 2003年第2期12卷 106-111页

作者：沈建英俞庆声王琪李泉蒲小平北京大学药学院分子与细胞药理学系 100083

Aim To analyze the secondary structure and neurotrophic effect of a specific protein in sensory neurons. Methods Comparison of the proteins expressed in the rat spinal sensory neurons and motor neurons was made by two dimensional electrophoresis. One specific protein in sensory neurons was isolated and purified by DEAE Sephacel ion exchange chromatography and high performance liquid chromatography. A primary analysis of its secondary structure by circular dichroism, and its neurotrophic effects were investigated using the model of dorsal root ganglia(DRG) cultured in vitro . Results The molecular weight and isoelectric point of the protein were 33 1 kDa and 5 52, respectively. Its circular dichroism showed that there were 20 8% α helix, 54 8% β sheet, 7 3% turn, and 17 1% random coil in its secondary structure. Biological experiments showed that the protein could promote the neurite outgrowth of DRG. Conclusion A specific protein in spinal sensory tissue with molecular weight of 33 1 kDa has been purified. There is mainly β sheet in the secondary structure of the protein. And the protein has neurotrophic effects in the model of DRG.

关键词： spinal sensory specific 33 1 kDa protein two dimensional electrophoresis high performance liquid chromatography circular dichroism neurotrophic effect