于常海
神经细胞
个性化签名
- 姓名:于常海
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学术头衔:
博士生导师
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学科领域:
病理学
- 研究兴趣:神经细胞
于常海教授,多年来主要从事神经细胞的研究,利用原代培养的神经细胞在分子和基因水平研究其损伤和再生。于常海教授在细胞培养、损伤模型的建立以及细胞神经化学方面具有非常丰富的经验。他早期的关于谷氨酸代谢的研究阐明了谷氨酸在脑中代谢的途径。他首次在厌氧箱内对细胞进行低密度培养,解决了神经科学界长期无法找到合适的体外缺血模型的问题。在1992年他首创了原代培养的星形胶质细胞的划伤模型,以此模拟中枢神经系统的机械损伤。这些工作为深入研究脑损伤和修复奠定了方法学基础,并在国际上广为应用。于常海教授是纽约科学院、美国神经化学协会、国际脑血流及代谢协会、神经科学协会和国际神经化学协会的会员,并在其中起骨干作用。在过去的10多年间,他先后五十多次受邀在各种国际科学学术会议上发表专题演讲。他积极致力于推动中国神经科学的国际交流,并多次组织在中国召开国际性神经科学会议;与此同时他还担任数个学术刊物,包括《中华医学杂志英文版》、《中国神经科学杂志》、《生理科学进展》、《Neurochemical Research》的编委。已出版英文编著2本。他共在国际性学术刊物发表论文一百余篇。并于2002-2003连续两年获得北京大学SCI优秀论文三等奖。
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【期刊论文】Nucleic acid sequence-based amplification methods to detect avian influenza virus
于常海
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-1年11月30日
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【期刊论文】A real-time PCR for SARS-coronavirus incorporating target gene pre-amplification
于常海
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-1年11月30日
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于常海, Albert Cheung Hoi Yu, , *, Hon Wa Yung, Michael Hung Kit Hui, Lok Ting Lau, Xiao Qian Chen, and Richard A. Collins
Journal of Neuroscience Research 74:318-325 (2003),-0001,():
-1年11月30日
An in vitro ischemia model was established and the effect of the metabolic inhibitors cycloheximide (CHX) and actinomycin D (ActD) on apoptosis in astrocytes under ischemia studied. CHX decreased by 75% the number of cells dying after 6 hr of ischemia compared with control cultures. TdT-mediated dUTP nick end labelling (TUNEL) staining of comparable cultures was reduced by 40%. ActD decreased cell death by 60% compared with controls. The number of TUNEL-positive cells was reduced by 38%. The nuclear shrinkage in TUNEL-positive astrocytes in control cultures did not occur in ActD-treated astrocytes, indicating that nuclear shrinkage and DNA fragmentation during apoptosis are two unrelated processes. Expression of bcl-2 (α and β), bax, and Ice in astrocytes under similar ischemic conditions, as measured by quantitative reverse transcription-polymerase chain reaction, indicated that ischemia down-regulated bcl-2 (α and β) and bax. Ice was initially down-regulated from 0 to 4 hr, before returning to control levels after 8 hr of ischemia. ActD decreased the expression of these genes. CHX reduced the expression of bcl-2 (α and β) but increased bax and Ice expression. It is hypothesized that the balance of proapoptotic (Bad, Bax) and antiapoptotic (Bcl-2, Bcl-Xl) proteins determines apoptosis. The data suggest that the ratio of Bcl-2/Bad in astrocytes following ActD and CHX treatment does not decrease as much in untreated cells during ischemia. Our data indicate that it is the ratio of Bcl-2 family members that plays a critical role in determining ischemia-induced apoptosis. It is also important to note that ischemia-induced apoptosis involves the regulation of RNA and protein synthesis.
actinomycin D, apoptosis, astrocytes, cycloheximide, Bcl-2, Bax, Ice, ischemia
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于常海, XIAO QIAN CHEN, , JIAN GUO CHEN, YUN ZHANG, WENDY WEN LUAN HSIAO, AND ALBERT CHEUNG HOI YU, *
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-1年11月30日
An in vitro ischemia model was established and the effect of the metabolic inhibitors cycloheximide (CHX) and actinomycin D (ActD) on apoptosis in astrocytes under ischemia studied. CHX decreased by 75% the number of cells dying after 6 hr of ischemia compared with control cultures. TdT-mediated dUTP nick end labelling (TUNEL) staining of comparable cultures was reduced by 40%. ActD decreased cell death by 60% compared with controls. The number of TUNEL-positive cells was reduced by 38%. The nuclear shrinkage in TUNEL-positive astrocytes in control cultures did not occur in ActD-treated astrocytes, indicating that nuclear shrinkage and DNA fragmentation during apoptosis are two unrelated processes. Expression of bcl-2 (α and β), bax, and Ice in astrocytes under similar ischemic conditions, as measured by quantitative reverse transcription-polymerase chain reaction, indicated that ischemia down-regulated bcl-2 (α and β) and bax. Ice was initially down-regulated from 0 to 4 hr, before returning to control levels after 8 hr of ischemia. ActD decreased the expression of these genes. CHX reduced the expression of bcl-2 (α and β) but increased bax and Ice expression. It is hypothesized that the balance of proapoptotic (Bad, Bax) and antiapoptotic (Bcl-2, Bcl-Xl) proteins determines apoptosis. The data suggest that the ratio of Bcl-2/Bad in astrocytes following ActD and CHX treatment does not decrease as much in untreated cells during ischemia. Our data indicate that it is the ratio of Bcl-2 family members that plays a critical role in determining ischemia-induced apoptosis. It is also important to note that ischemia-induced apoptosis involves the regulation of RNA and protein synthesis.
RNA arbitrarily primed PCR, differential gene expression, injury, LY294002, U0126
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【期刊论文】Central Nervous System and Limb Anomalies in Case Reports of First-Trimester Statin Exposure
于常海
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-1年11月30日
The 14-3-3 protein family comprises critical regulatory molecules involved in signaling during cell division, proliferation, and apoptosis. Despite extensive study, the functions of the 14-3-3 proteins in brain remain unclear. 14-3-3γ, a subtype of the 14-3-3 family of proteins, was thought to be brain-and neuron-specific. Using RNA arbitrarily primed PCR, we identified an upregulated cDNA fragment of the 14-3-3γ gene in primary cultures of astrocytes. Using Northern blot analysis, we confirmed this fragment was brain-specific. In cultures of astrocytes, 14-3-3γgenes and proteins were differentially expressed at different ages and the proteins were distributed only in the cytoplasm. These results indicated that 14-3-3 was not neuron-specific but also expressed in astrocytes. The function of this protein in brain is unclear. Northern and Western blot analyses demonstrated that 14-3-3γmRNA and protein were upregulated in cultured astrocytes in an anaerobic chamber-induced ischemia model. The induction of 14-3-3γproteins was neither suppressed by an MAP kinase inhibitor (U0126) nor a PI-3 kinase inhibitor (LY294002). These data indicated that induction of 14-3-3γmight not involve PI-3 and MAP kinase-dependent pathways. Using coimmunoprecipitation, we demonstrated that endogenous 14-3-3γbound to c-Raf-1 and p-Raf 259. As Raf is one of the critical serine/threonine kinases controlling cell growth, differentiation, and death, the binding of 14-3-3γto Raf indicates the critical role of this protein in ischemiainduced apoptosis and the changes in signal transduction in astrocytes in culture. GLIA 42: 315-324, 2003.
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【期刊论文】Rapid and sensitive detection of avian influenza virus subtype H7 using NASBA
于常海, Richard A. Collins, a Lung-Sang Ko, a King-Yip Fung, a Ka-Yun Chan, a Jun Xing, a Lok-Ting Lau, a, b and Albert Cheung Hoi Yua, b, *
Biochemical and Biophysical Research Communications 300 (2003) 507-515,-0001,():
-1年11月30日
The Six Cities Study by Dockery et al.1 has captured renewed attention (Jan. 8 issue).2,3 Claiming to identify an increased mortality rate in Steubenville, Ohio, as compared with the rate in Portage, Wisconsin, the authors posited that air pollution in Steubenville, an Ohio River Valley mining and steel-making city, was responsible for its increased death rate. That interpretation is not permissible, inasmuch as the socioeconomic and ethnic makeups of the populations of Portage and Steubenville are markedly different. Compounding that invidious comparison, the calculated difference in mortality rates is seen to be only a hair’s breadth within the bounds of statistical significance. Indeed, as shown in Table 2 of the report,1 there was no difference in mortality between women in Portage and women in Steubenville, who breathed the same air as men in that city. Steinbrook3 quotes David M. Michaels, formerlyformerly of the Clinton administration, who suggests that critics of clean-air studies are “hired guns working for dirty companies.” Joel Schwartz, later associated with the clean-air group at Harvard, reportedly dismissed critics of clean-air studies as “industry thugs.”4 Is this the language of collegial scientific discourse?
NASBA, Avian influenza, H7, Haemagglutinin, Electrochemiluminescence
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【期刊论文】Comparison of nucleic acid-based detection of avian influenza H5N1 with virus isolation
于常海, Songhua Shan, a Lung-Sang Ko, b Richard A. Collins, b Zhongliang Wu, a Jiahua Chen, a Ka-Yun Chan, b Jun Xing, b Lok-Ting Lau, b, c and Albert Cheung-Hoi Yub, c, *
Biochemical and Biophysical Research Communications 302 (2003) 377-383,-0001,():
-1年11月30日
Nucleic acid sequence-based amplification with electrochemiluminescent detection (NASBA/ECL) is an isothermal technique allowing rapid amplification and detection of specific regions of nucleic acid from a diverse range of sources. It is especially suitable for amplifying RNA. A NASBA/ECL technique has been developed allowing the detection of RNA from avian influenza virus subtype H7 derived from allantoic fluid harvested from inoculated chick embryos and from cell cultures. Degenerate amplification primers and amplicon capture probes were designed enabling the detection of low and highly pathogenic avian influenza of the H7 subtype from the Eurasian and North American lineages and the Australian sub-lineage. The NASBA/ECL technique is specific for subtype H7 and does not cross-react with other influenza subtypes or with viruses containing haemagglutinin-like genes. The assay is 10- to 100-fold more sensitive than a commercially available antigen capture immunoassay system. The NASBA/ECL assay could be used in high throughput poultry screening programmes.
Highly pathogenic avian influenza, Nucleic acid sequence-based amplification, H5N1, Electrochemiluminescence, Virus isolation
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【期刊论文】Genome comparison of a novel foot-and-mouth disease virus with other FMDV strains
于常海, Qian Fenga, Huan Yub, Yingying Liua, Chengqiang Hea, Jingsong Hua, Huachun Sanga, Neizheng Dinga, Mingxiao Dinga, Yin-Wan Wendy Fungc, e, Lok-Ting Lauc, Albert Cheung-Hoi Yuc, Jianguo Chena, d, *
Biochemical and Biophysical Research Communications 323 (2004) 254-263,-0001,():
-1年11月30日
Nucleic acid sequence-based amplification with electrochemiluminescent detection (NASBA/ECL) of avian influenza virus was compared with viral culture in embryonated chicken eggs. Virus was isolated from blood or anal swabs of chickens artificially infected with highly pathogenic avian influenza A/Chicken/Hong Kong/1000/97 (H5N1). Viral nucleic acid was detected in blood samples by NASBA/ECL immediately prior to death, whilst nucleic acid extracted from anal swabs was detected from the day following artificial infection until death. Thus, blood and/or anal swabs are a suitable source of material for the detection of avian influenza in dead birds, but anal swabs are more suitable for detection of viral genetic material in live birds. Dilution of a known viral standard was used to determine the limit of sensitivity for both NASBA/ECL and egg culture detection methods. The NASBA/ ECL method was equivalent in sensitivity to egg culture. The NASBA/ECL results agreed with egg culture data in 71/94 (75.5%) tissue samples obtained from artificially infected birds.
Picornavirus, Serotype, Phylogenesis, Sequence, Alignment
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【期刊论文】The necessity of molecular diagnostics for avian flu
于常海, Yin-Wan Wendy Fung, Lok-Ting Lau & Albert Cheung-Hoi Yu
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-1年11月30日
The genome of a novel foot-and-mouth disease virus, HKN/2002, was 8104 nucleotides (nt) in length (excluding the poly(C) tract and poly(A) tail) and was composed of a 1042-nt 50-untranslated region (UTR), a 6966-nt open reading frame, and a 93-nt 30-UTR. Genome sequences of HKN/2002 and other known FMDV strains were compared. The VP1, VP2, and VP3-based neighbor-joining (NJ) trees were divided into distinct clusters according to different serotypes, while other region-based NJ trees exhibited some degree of intercross among serotypes. Mutations in HKN/2002 were revealed, including frequent deletions and insertions in the G-H loop of VP1, and deletion involving 10 amino acid residues in the 3A protein. An evolutionary relationship of HKN/2002 with an Asian FMDV lineage isolated from a Hong Kong swine host in 1970 was postulated. A 43-nt deletion identified in the 50-UTR of HKN/2002 possibly contributed to the loss of one pseudo-knot domain.
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