Monkey B virus(Macacine herpesvirus 1; BV) is noted for its extreme neurovirulence in humans. Since the vhs protein encoded by the UL41 gene has been shown to be a neurovirulence factor in the related human herpes sim...Monkey B virus(Macacine herpesvirus 1; BV) is noted for its extreme neurovirulence in humans. Since the vhs protein encoded by the UL41 gene has been shown to be a neurovirulence factor in the related human herpes simplex viruses, the role of the UL41 gene in BV neurovirulence was investigated. BV mutants were constructed that lacked the entire UL41 ORF(Δ41) or had the RNase active site mutated(Δ41A). Neither mutant shut off host protein synthesis, degraded β-actin mRNA, or prevented an IFN-β response, indicating that the vhs protein and its RNase activity are both necessary for these activities. Replication of both mutants in primary mouse cells was impaired and they exhibited a prolonged disease course in mice. Whereas Δ41 infected mice were euthanized for symptoms related to central nervous system(CNS) infection, Δ41A infected mice were euthanized primarily for symptoms of autonomic nervous system dysfunction. While neuroinvasiveness was not affected, lesions in the CNS were more limited in size, anatomical distribution, and severity than for wild-type virus. These results indicate that the vhs protein affects the general replicative efficiency of BV in vivo rather than being a specific neurovirulence factor critical for invasion of or preferential replication in the CNS.展开更多
Large DNA viruses normally have complex structures with many of protein components derived from both viral and host origins. The development in proteomics, especially mass spectrometry identification techniques provid...Large DNA viruses normally have complex structures with many of protein components derived from both viral and host origins. The development in proteomics, especially mass spectrometry identification techniques provide powerful tools for analyzing large viruses. In this review, we have summarized the recent achievements on proteomic studies of large DNA viruses, such as herpesvirus, poxvirus, nimavirus and baculoviruse. The proteomics of baculovirus occlusion-derived virions (ODV) were emphasized. Different mass spectrometry techniques used on various baculoviruses were introduced, and the identified structurally associated proteins of baculoviruses are summarized.展开更多
Wheat rosette stunt virus (WRSV) contains an RNA-dependent RNA polymerase. The RNA polymerase activity is associated with the viral nucleocapsid (NP).In vitro transcription of purified NP required all of the four nucl...Wheat rosette stunt virus (WRSV) contains an RNA-dependent RNA polymerase. The RNA polymerase activity is associated with the viral nucleocapsid (NP).In vitro transcription of purified NP required all of the four nucleoside triphosphates and Mg^(2+).There was a need of a proper salt concentration and some reducing reagents in the system for increasing the RNA polymerase activity.The optimum temperature for in vitro transcription was around 25℃.Within the first 90 min of in vitro reaction, incorporation rose linearly with the time course of incubation. The experiments of ribonuclease and deoxyribonuclease treatments showed that single-stranded RNAs were synthesized in vitro by the RNA polymerase.Two fractions of WRSV-NP could be separated by SDS-dissociation and ultracentrifugation.The supernatant fraction contained three structural proteins of NP: L, N and NS;and the pellet fraction contained the viral RNA.When the supernatant proteins and the viral RNA were mixed together, RNA polymerase activity could be reconstituted.When the ratio between the amounts of the supernatant proteins and the viral RNA in the mixture was about 100:7.7, the reconstituted RNA polymerase activity reached the maximum.展开更多
Discrete computer simulations are quite helpful in understanding dynamic structures in complex systems.Recently,using the Mole-8.5 supercomputer and molecular dynamics simulations as a"computational microscope&qu...Discrete computer simulations are quite helpful in understanding dynamic structures in complex systems.Recently,using the Mole-8.5 supercomputer and molecular dynamics simulations as a"computational microscope",we simulated the dynamic structure of a whole H1N1 influenza virion in solution for the first time at the atomic level.In total,300 million atoms in a periodic cube with an edge length of 148.5 nm were simulated.Using 288 low level hybrids with 1728 C2050 GPUs and a software package developed specifically for the hardware,the simulation executed 770 ps/d with an integration time step of 1 fs,and analyzed the dynamic structure.With the tremendous computational power of GPUs,efficient software packages for various hardware designs,and consistent physical models,more challenging applications will be carried out in the near future.展开更多
Objective: To explore the effect of morphological changes in the development of condyloma acuminatum (CA) Materials and Methods: Lesions in five patients with CA were observed . Results: Upon electron microscopy, the ...Objective: To explore the effect of morphological changes in the development of condyloma acuminatum (CA) Materials and Methods: Lesions in five patients with CA were observed . Results: Upon electron microscopy, the most characteristic feature of the lesions important for diagnosis, was the presence of distinct perinuclear vacuolizations, or so-called koilocytes, among some epithelial cells. These cells possessed hyperchromatic nuclei, swollen mitochondria, dilated endoplasmic reticulum and dissolved glycogen. There were interchromatin granules and perichromatin granules in some nuclei. Moreover some virus particles were also seen in the nuclei of some infected cells. Conclusions: The ultrastructural findings may be used to histopathologically explain the pathogenesis and mechanism of this disease, and it is helpful for diagnosis of CA .展开更多
While host proteins incorporated into virions during viral budding from infected cell are known to play essential roles in multiple process of the life cycle of progeny virus,these characteristics have been largely ne...While host proteins incorporated into virions during viral budding from infected cell are known to play essential roles in multiple process of the life cycle of progeny virus,these characteristics have been largely neglected in studies on rabies virus(RABV).Here,we purified the RABV virions with good purity and integrity,and analyzed their proteome by nano LC–MS/MS,followed by the confirmation with immunoblot and immuno-electronic microscopy.In addition to the 5 viral proteins,49 cellular proteins were reproducibly identified to be incorporated into matured RABV virions.Function annotation suggested that 24 of them were likely involved in virus replication.Furthermore,cryo-EM was employed to observe the purified RABV virions,generating high-resolution pictures of the bullet-shaped virion structure of RABV.This study has provided new insights into the host proteins composition in RABV virion and shed the light for further investigation on molecular mechanisms of RABV infection,as well as the discovery of new anti-RABV therapeutics.展开更多
目的观察国产成人流感裂解疫苗的安全性。方法对2013年四川省内3岁及3岁以上人群接种长春生物制品研究所有限责任公司(简称长春公司)生产的成人流感裂解疫苗以及其余所有厂家的流感裂解疫苗(对照流感疫苗)后报告的疑似预防接种异常反应(...目的观察国产成人流感裂解疫苗的安全性。方法对2013年四川省内3岁及3岁以上人群接种长春生物制品研究所有限责任公司(简称长春公司)生产的成人流感裂解疫苗以及其余所有厂家的流感裂解疫苗(对照流感疫苗)后报告的疑似预防接种异常反应(adverse events following immunization,AEFI)情况进行回顾性调查。结果2013年四川省接种长春公司流感裂解疫苗后共报告AEFI 36例,报告发生率为11.78/10万,其中一般反应发生率为8.5/10万,异常反应发生率为2.62/10万。与对照流感疫苗相比,长春公司流感疫苗的大年龄组别不良反应构成比较高。接种长春公司流感疫苗后,一般反应(发热、硬结、红肿)的发生率低于对照流感疫苗;除荨麻疹外,其余异常反应的报告发生率均低于对照流感疫苗;疫苗的异常反应无批号聚集性。结论长春公司生产的成人流感裂解疫苗安全性较好。展开更多
基金supported in part by PHS grants 2P40 OD010988 and 1P40 OD010431
文摘Monkey B virus(Macacine herpesvirus 1; BV) is noted for its extreme neurovirulence in humans. Since the vhs protein encoded by the UL41 gene has been shown to be a neurovirulence factor in the related human herpes simplex viruses, the role of the UL41 gene in BV neurovirulence was investigated. BV mutants were constructed that lacked the entire UL41 ORF(Δ41) or had the RNase active site mutated(Δ41A). Neither mutant shut off host protein synthesis, degraded β-actin mRNA, or prevented an IFN-β response, indicating that the vhs protein and its RNase activity are both necessary for these activities. Replication of both mutants in primary mouse cells was impaired and they exhibited a prolonged disease course in mice. Whereas Δ41 infected mice were euthanized for symptoms related to central nervous system(CNS) infection, Δ41A infected mice were euthanized primarily for symptoms of autonomic nervous system dysfunction. While neuroinvasiveness was not affected, lesions in the CNS were more limited in size, anatomical distribution, and severity than for wild-type virus. These results indicate that the vhs protein affects the general replicative efficiency of BV in vivo rather than being a specific neurovirulence factor critical for invasion of or preferential replication in the CNS.
基金The grants of National Science Foundation of China (30630002, 30670078)973 Program(2009CB118903)Programme Strategic Scientific Alliances between China and the Netherlands (2008AA000238)
文摘Large DNA viruses normally have complex structures with many of protein components derived from both viral and host origins. The development in proteomics, especially mass spectrometry identification techniques provide powerful tools for analyzing large viruses. In this review, we have summarized the recent achievements on proteomic studies of large DNA viruses, such as herpesvirus, poxvirus, nimavirus and baculoviruse. The proteomics of baculovirus occlusion-derived virions (ODV) were emphasized. Different mass spectrometry techniques used on various baculoviruses were introduced, and the identified structurally associated proteins of baculoviruses are summarized.
基金Project supported by the National Natural Science Foundation of China.
文摘Wheat rosette stunt virus (WRSV) contains an RNA-dependent RNA polymerase. The RNA polymerase activity is associated with the viral nucleocapsid (NP).In vitro transcription of purified NP required all of the four nucleoside triphosphates and Mg^(2+).There was a need of a proper salt concentration and some reducing reagents in the system for increasing the RNA polymerase activity.The optimum temperature for in vitro transcription was around 25℃.Within the first 90 min of in vitro reaction, incorporation rose linearly with the time course of incubation. The experiments of ribonuclease and deoxyribonuclease treatments showed that single-stranded RNAs were synthesized in vitro by the RNA polymerase.Two fractions of WRSV-NP could be separated by SDS-dissociation and ultracentrifugation.The supernatant fraction contained three structural proteins of NP: L, N and NS;and the pellet fraction contained the viral RNA.When the supernatant proteins and the viral RNA were mixed together, RNA polymerase activity could be reconstituted.When the ratio between the amounts of the supernatant proteins and the viral RNA in the mixture was about 100:7.7, the reconstituted RNA polymerase activity reached the maximum.
基金supported by Ministry of Finance(ZDYZ2008-2)the National Key Science and Technology Project(2008ZX05014-003-006HZ)+1 种基金the Knowledge Innovation Project of Chinese Academy of Sciences (KGCX2-YW-124)the National Natural Science Foundation of China (20821092)
文摘Discrete computer simulations are quite helpful in understanding dynamic structures in complex systems.Recently,using the Mole-8.5 supercomputer and molecular dynamics simulations as a"computational microscope",we simulated the dynamic structure of a whole H1N1 influenza virion in solution for the first time at the atomic level.In total,300 million atoms in a periodic cube with an edge length of 148.5 nm were simulated.Using 288 low level hybrids with 1728 C2050 GPUs and a software package developed specifically for the hardware,the simulation executed 770 ps/d with an integration time step of 1 fs,and analyzed the dynamic structure.With the tremendous computational power of GPUs,efficient software packages for various hardware designs,and consistent physical models,more challenging applications will be carried out in the near future.
基金Funded by the Natural Science Foundation of Heilongjiang,China.
文摘Objective: To explore the effect of morphological changes in the development of condyloma acuminatum (CA) Materials and Methods: Lesions in five patients with CA were observed . Results: Upon electron microscopy, the most characteristic feature of the lesions important for diagnosis, was the presence of distinct perinuclear vacuolizations, or so-called koilocytes, among some epithelial cells. These cells possessed hyperchromatic nuclei, swollen mitochondria, dilated endoplasmic reticulum and dissolved glycogen. There were interchromatin granules and perichromatin granules in some nuclei. Moreover some virus particles were also seen in the nuclei of some infected cells. Conclusions: The ultrastructural findings may be used to histopathologically explain the pathogenesis and mechanism of this disease, and it is helpful for diagnosis of CA .
基金This research was funded by the National Key Research and Development Plan(Grant No.2016YFD0500401)National Natural Science Foundation of China(Grant No.31402214)China Postdoctoral Science Foundation(Grant No.2014M552638).
文摘While host proteins incorporated into virions during viral budding from infected cell are known to play essential roles in multiple process of the life cycle of progeny virus,these characteristics have been largely neglected in studies on rabies virus(RABV).Here,we purified the RABV virions with good purity and integrity,and analyzed their proteome by nano LC–MS/MS,followed by the confirmation with immunoblot and immuno-electronic microscopy.In addition to the 5 viral proteins,49 cellular proteins were reproducibly identified to be incorporated into matured RABV virions.Function annotation suggested that 24 of them were likely involved in virus replication.Furthermore,cryo-EM was employed to observe the purified RABV virions,generating high-resolution pictures of the bullet-shaped virion structure of RABV.This study has provided new insights into the host proteins composition in RABV virion and shed the light for further investigation on molecular mechanisms of RABV infection,as well as the discovery of new anti-RABV therapeutics.
文摘目的观察国产成人流感裂解疫苗的安全性。方法对2013年四川省内3岁及3岁以上人群接种长春生物制品研究所有限责任公司(简称长春公司)生产的成人流感裂解疫苗以及其余所有厂家的流感裂解疫苗(对照流感疫苗)后报告的疑似预防接种异常反应(adverse events following immunization,AEFI)情况进行回顾性调查。结果2013年四川省接种长春公司流感裂解疫苗后共报告AEFI 36例,报告发生率为11.78/10万,其中一般反应发生率为8.5/10万,异常反应发生率为2.62/10万。与对照流感疫苗相比,长春公司流感疫苗的大年龄组别不良反应构成比较高。接种长春公司流感疫苗后,一般反应(发热、硬结、红肿)的发生率低于对照流感疫苗;除荨麻疹外,其余异常反应的报告发生率均低于对照流感疫苗;疫苗的异常反应无批号聚集性。结论长春公司生产的成人流感裂解疫苗安全性较好。
