秦启伟
鱼类细胞和分子免疫学,海洋分子病毒学,基因组学和蛋白质组学,基因芯片,微芯片实验室(Lab-on-chip)以及流式细胞技术(Flow Cytometry)在海洋环境微生物检测中的应用
个性化签名
- 姓名:秦启伟
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学术头衔:
博士生导师
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学科领域:
海洋生物学
- 研究兴趣:鱼类细胞和分子免疫学,海洋分子病毒学,基因组学和蛋白质组学,基因芯片,微芯片实验室(Lab-on-chip)以及流式细胞技术(Flow Cytometry)在海洋环境微生物检测中的应用
秦启伟博士为中山大学"百人计划"引进人才,中山大学生命科学学院,生物防治国家重点实验室教授,博士生导师,专业为海洋生物学及生化与分子生物学,研究方向为海洋分子微生物学及功能基因组学.秦教授曾作为课题组长(PI)或实验室主任在国内外多个著名的海洋研究机构从事海洋生物学的研究及技术开发工作.近年来,在鱼类细胞和分子免疫学,海洋分子病毒学,基因组学和蛋白质组学,基因芯片,微芯片实验室(Lab-on-chip)以及流式细胞技术(Flow Cytometry)在海洋环境微生物检测中的应用等方面,取得了一系列学术成果.近五年来(2000-2004),在国内外重要学术期刊上已发表论文20多篇,其中SCI收录15篇.
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秦启伟, 吴灶和, 周永灿, 潘金培
热带海洋,2000,19(1):58~63,-0001,():
-1年11月30日
调查了饵料维生素C对青石斑鱼Epinephelus awoara非特异性的体液和细胞免疫调节作用。在冰冻饵料小杂鱼中添加不同剂量的维生素C(每公斤饵料鱼含量分别为0,500,1000,1500和2000mg),连续投喂青石斑鱼20周后,青石斑鱼血液白细胞总数及不同种类白细胞(包括淋巴细胞、中性粒细胞和单核细胞)组成比例不受饵料维生素C的影响。青石斑鱼血清补体经典型溶解羊红细胞的能力随饵料维生素C添加量的提高而显著增强(p<0.01)。然而,维生素C对血清的杀菌活性没有作用。鱼体非特异性的细胞免疫活动(包括血液白细胞和头肾细胞的吞噬活动)也不受维生素C的影响。
青石斑鱼Epinephelus awoara, 维生素C, 免疫刺激剂, 非特异性免疫
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【期刊论文】Production and characterization of monoclonal antibodies to a grouper iridovirus
秦启伟, Chengyin Shi a, , Qi Wei a, b, *, Karina Yew Hoong Gin c, Toong Jin Lam a
Journal of Virological Methods 107(2003)147-154,-0001,():
-1年11月30日
A panel of six monoclonal antibodies (mAbs) against a grouper iridovirus (SGIV) were produced by immunization of Balb/c mice with purified virus preparations. Isotyping test revealed all the mAbs were IgG1. None of the mAbs possessed the ability to neutralize SGIV in cell cultures. Western blot showed that 4mAbs reacted with 2 SGIV proteins at molecular mass of approximately 100 and 117kDa in gradient-purified virus. Immunofluorescent studies showed that the two specific viral proteins VP100 and VP117 were localized within virus assembly sites in the cytoplasm of SGIV-infected grouper cells (GP). Fractionations of the iridovirus in a 20-60% sucrose gradient were detected successfully by all the six mAbs using immunodot blot. An antigen-capture enzyme-linked immunosorbent assay (ELISA) system, based on the use of mAb 7E11 for capture and a rabbit polyclonal antibody to SGIV for detection was developed. SGIV antigen was detected in gradient-purified virus and virus-infected grouper blood. These novel mAbs will facilitate the development of more specific and standardized diagnostic techniques for marine fish iridovirus.
Monoclonal antibodies, Grouper, Epinephelus spp., , Iridovirus
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秦启伟, Canhua Huang a, Xiaobo Zhang b, Karina Y.H. Gin c, Qi Wei Qin d, e, *
Journal of Virological Methods 117(2004)123-128,-0001,():
-1年11月30日
A DNA probe of 531 base pairs for Singapore grouper iridovirus (SGIV) was generated by polymerase chain reaction and labeled with nonradioactive digoxigenin. An in situ hybridization based method was developed to detect SGIV in formalin-fixed tissues from maricultured Malabar grouper, Epinephelus malabaricus Bloch and Schneider. The in situ hybridization detected SGIV in the kidney, spleen, liver, intestine, stomach and gills from naturally infected fish. Strong hybridization signals were obtained from the kidney and spleen tissues, while intermediate intensity signals were observed in the intestine and liver tissues. The weakest signals were obtained from the stomach and gills. The signals were located specifically within epithelial, endothelial and sub-endothelial hypertrophic cells in all tested tissues. The in situ hybridization procedure will provide an important diagnostic tool to complement histopathological methods, and contribute to epidemiological studies on the origin and distribution of iridovirus in mariculture.
DNA probe, In situ hybridization, Iridovirus, Grouper
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【期刊论文】Graft-versus-host reaction (GVHR) in clonal amago salmon, Oncorhynchus rhodurus
秦启伟, Q.W. Qin a, *, M. Ototake b, H. Nagoya b, T. Nakanishi c
Veterinary Immunology and Immunopathology 89(2002)83-89,-0001,():
-1年11月30日
The graft-versus-host reaction (GVHR) was demonstrated in a salmonid model system of clonal diploid and triploid amago salmon. Triploid operculum grafts on clonal diploid evoked an acute rejection within 12 days. Grafts exchanged among triploid amago salmon exhibited prolonged survival for 18 days. In contrast, diploid grafts on triploid, and allografts among clonal diploid amago salmon were accepted. A typical GVHR was induced in triploid recipients by intraperitonal injection of head kidney cells from sensitised diploid donors. The clinical signs of graft-versus-host disease (GVHD) were observed in the recipients after 1 week of cell injection as a loss of appetite and appearance of solid faeces, followed by haemorrhage, local swelling of ventral skin and an enlarged spleen. Three of six fish died within 1 month. Water temperature and frequency of sensitisation are critical to induce GVHR. Diploid donors had to be sensitised three times at 20 8C to induce the typical GVHR. GVHR was most effectively induced by head kidney cells, followed by peripheral blood leucocytes (PBL) and spleen cells. Ploidy analysis by flow cytometry revealed that the donor head kidney cells greatly increased in the recipient liver, head kidney and spleen, and reached the peak after 9 days of donor cell injection. The results in the present study are quite similar to the findings in ginbuna and ginbuna-gold fish hybrid system, suggesting the presence of T cells in salmonid as well as cyprinid fish.
Graft-versus-host reaction (, GVHR), , Clonal amago salmon, Oncorhynchus rhodurus, Graft-versus-host disease (, GVHD), , Cell-mediated immunity
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秦启伟, Wen Jun Song, Qi Wei Qin, Jin Qiu, Can Hua Huang, Fan Wang, and Choy Leong Hew , *
JOURNAL OF VIROLOGY, Nov. 2004, p. 12576-12590,-0001,():
-1年11月30日
Here we report the complete genome sequence of Singapore grouper iridovirus (SGIV). Sequencing of the random shotgun and restriction endonuclease genomic libraries showed that the entire SGIV genome consists of 140,131 nucleotide bp. One hundred sixty-two open reading frames (ORFs) from the sense and antisense DNA strands, coding for lengths varying from 41 to 1,268 amino acids, were identified. Computer-assisted analyses of the deduced amino acid sequences revealed that 77 of the ORFs exhibited homologies to known virus genes, 23 of which matched functional iridovirus proteins. Forty-two putative conserved domains or signatures were detected in the National Center for Biotechnology Information CD-Search database and PROSITE database. An assortment of enzyme activities involved in DNA replication, transcription, nucleotide metabolism, cell signaling, etc., were identified. Viruses were cultured on a cell line derived from the embryonated egg of the grouper Epinephelus tauvina, isolated, and purified by sucrose gradient ultracentrifugation. The protein extract from the purified virions was analyzed by polyacrylamide gel electrophoresis followed by in-gel digestion of protein bands. Matrix-assisted laser desorption ionization-time of flight mass spectrometry and database searching led to identification of 26 proteins. Twenty of these represented novel or previously unidentified genes, which were further confirmed by reverse transcription-PCR (RT-PCR) and DNA sequencing of their respective RT-PCR products.
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秦启伟, Q.W. Qin a, *, T.J. Lam a, b, Y.M. Sin b, H. Shen a, S.F. Chang c, G.H. Ngoh c, C.L. Chen b
Journal of Virological Methods 98(2001)17-24,-0001,():
-1年11月30日
The morphogenesis and the ultrastructure of a marine fish iridovirus isolated from diseased grouper, Epinephelus tau ina were studied by electron microscopy. The virus was grown on a marine fish cell line (GP) at 25℃. After appearance of advanced cytopathic effect (CPE), various morphogenetic stages of virus amplification, maturation and assembly were detected in the cytoplasm of virus-infected cells. The matured nucleocapsids were probably formed by insertion of electron-dense core material into a partly forming empty capsid just before completely sealed. The nucleocapsids were located at the assembly sites as pseudocrystalline arrays or scattered individually. In the late phase of infection, the nucleocapsids were enveloped and released by budding from the plasma membrane. The budding virus particles could directly enter neighbouring cells by endocytosis to start the next round infection. Ultrastructure of the grouper iridovirus was studied using the methods of enzymatic digestions and detergent degradations. The purified iridovirus particles showed a three-layered membrane including an external lipoprotein envelope, an inner periodic protein capsid and a lipid-containing membrane. The regular array of surface capsid subunits was observed after degradation with detergent.
Morphogenesis, Ultrastructure, Iridovirus, Grouper, Epinephelus tauvina
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秦启伟, QIN Qi-wei>, >, WU Zao-he>, PAN Jin-pei>
Chinese Journal of Oceanology and Limnology Vol. 18, No.3, P. 247-252, 2000,-0001,():
-1年11月30日
The disease resistance and humoral immunomodulatory effects of vitamin C administered orally to grouper, Epinephelus awoara maintained on a frozen fish diet supplemented with vitamin C at 500, 1000, 1500 and 2000mg/kg were investigated. After 20 weeks, the growth rates of the groups with high level of vitamin C apparently increased. The untreated fish had symptoms of vitamin C deficiency. The endogenous liver tissue vitamin C levels were found to reflect well the dietary treatments. After intraperitoneal injection or bath challenge with a virulent strain of Vibrio vulnificus, fish fed with high level vitamin C showed significantly higher survival rate compared with the normal control group. Vaccination with formalin inactivated V. vulnificus significantly enhanced the specific antibody production in fish treated with vitamin C, and completely protected from strong bacterial challenge the groups fed on fish with vitamin C 1500 and 2000mg/kg diet.
grouper,, Epinephelus awoara,, vitamin C,, disease resistance,, immunomodulation,, pathogen of Vibrio vulnificus
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秦启伟, S.F. Chang a, ), G.H. Ngoh a, L.F.S. Kueh a, Q.W. Qin b, C.L. Chen c, T.J. Lam c, Y.M. Sin c
Aquaculture 192(2001)133-145,-0001,():
-1年11月30日
A tropical marine fish cell line SF.was established from Asian seabass Lates calcarifer.fry. The cell line was maintained in Earle's minimum essential medium EMEM.supplemented with foetal calf serum and incubated at 258C. The susceptibility of the cell-line at the 16th, 35th and 46th subculture to several iridoviruses, birnaviruses, reoviruses, a rhabdovirus, and a nodavirus was studied. Cytopathic effect CPE. was observed daily after virus inoculation and viral replication efficiency was determined for six viruses. SF cell cultures infected with an iridovirus, a nodavirus and a reovirus were further elucidated by electron microscopy. All the viruses tested were shown to induce CPE on SF cells. This suggests that the SF cell line has good potential for the isolation of various fish viruses. The SF cell line consists predominantly of epithelial-like cells and has been subcultured 85 times over a period of 24 months.
Lates calcarifer, Tropical marine fish cell line, Virus
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秦启伟, Qi Wei Qin a, b, *, Karina Yew-Hoong Gin c, Li Yen Lee b, Alice Ilaya Gedaria b, Sheng Zhang b
Journal of Virological Methods 125(2005)49-54,-0001,():
-1年11月30日
A sensitive and accurate flow cytometry (FCM) based method has been developed to detect and quantitate a novel marine fish iridovirus (Singapore grouper iridovirus, SGIV) after amplification in cell cultures. Confluent grouper cell (GP) monolayers were infected with SGIV. When advanced cytopathic effect (CPE) appeared, the cell cultures were fixed and permeabilized, and then reacted with monoclonal antibodies specific against SGIV, followed by a second antibody conjugated with FITC (anti-mouse IgG-FITC). A Coulter
Flow cytometry, Marine fish iridovirus (, SGIV), , Cell culture, Monoclonal antibody, Immunofluorescence
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【期刊论文】Antiviral properties of hemocyanin isolated from shrimp Penaeus monodon
秦启伟, Xiaobo Zhang a, Canhua Huang b, Qiwei Qin c, *
Antiviral Research 61(2004)93-99,-0001,():
-1年11月30日
Penaeid shrimp aquaculture has suffered from many diseases, especially from viral origin such as white spot syndrome virus (WSSV). In an attempt to obtain antiviral-relevant proteins, two peptides with molecular masses at 73 and 75kDa were isolated from shrimp Penaeus monodon using affinity chromatography coupled with the purified WSSV or a fish iridovirus (Singapore grouper iridovirus, SGIV), and identified as hemocyanin by mass spectrometry. The results, using fish viruses capable of cell culture, showed for the first time that the hemocyanin had non-specific antiviral properties and no cytotoxicity against host cells.
Shrimp, Penaeus monodon, Hemocyanin, Antivirus
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