Development and application of real-time qPCR assay for detecting Sendai virus gene copy number / 中华实验和临床病毒学杂志

Objective@#To develop a real-time quantitative PCR (qPCR) assay for detecting Sendai virus gene copy number.@*Methods@#The recombinant plasmid was constructed and a reference used to establish the real-time qPCR method with the TaqMan probe and verified for reproducibility and sensitivity.@*Results@#The linear range of the developed real-time qPCR assay was 1.024 × 103 ~5 × 1010 copies/μl, with an R2 value of more than 0. 99. The standard recovery of the tested samples was 84.56%~119.48%.@*Conclusions@#Compared with traditional hernagglutination assay for particle number detection, this method has higher sensitivity, better repeatability and high quantitative accuracy. It can be used for the detection of particle number of vaccine with sendai virus as the carrier, so as to detect the specific activity of vaccine products.

Induction of pluripotent stem cells by reprogramming human ocular fibroblasts under xeno-free conditions / Indução de células-tronco pluripotentes pela reprogramação de fibroblastos oculares humanos sob condições xeno-livre

ABSTRACT Purposes: To develop an efficient and xeno-free standard eye-derived induced pluripotent stem cell reprogramming protocol for use during induced pluripotent stem cell-based cell therapies in treating retinal degenerative diseases and to compare the relative effectiveness of both animal- and non-animal-derived culture systems in the generation of induced pluripotent stem cells. Methods: Primary cultured human pterygium fibroblasts and human Tenon's capsule fibroblasts were induced to induced pluripotent stem cells using a non-in­tegrated virus under two xeno-free systems; as part of this study, a traditional non-xeno-free reprogramming system was also assessed. Induced pluripotent stem cell clones were selected and counted by live staining. Reprogramming efficiencies were evaluated between the fibroblasts and among different culture systems. In a series of experiments, such as PCR and immunofluorescence staining, the induced pluripotent stem cells were characterized. Results: Human pterygium fibroblast- and human Tenon's capsule fibroblast-derived induced pluripotent stem cells were successfully established using different reprogramming systems, under which they exhibited properties of induced pluripotent stem cells. Reprogramming efficiencies of induced pluripotent stem cells using the cell therapy system, the traditional system, and the E6/E8 system were 0.014%, 0.028%, and 0.001%, respectively, and those of human pterygium fibroblast- and human Tenon's capsule fibroblast-derived induced pluripotent stem cells-using the aforementioned systems-were 0.018% and 0.017%, respectively. Conclusions: Sendai virus facilitates induced pluripotent stem cell reprogramming of ocular fibroblasts-both human pterygium and human Tenon's capsule fibroblasts being safe and efficient for induced pluripotent stem cell reprogramming. Although the reprogramming efficiencies of ocular-derived induced pluripotent stem cells under xeno-free conditions were not superior to those observed using the traditional reprogramming system, the cell therapy system reprogramming system is a good option when induced pluripotent stem cells are to be induced under xeno-free conditions.

RESUMO Objetivos: Desenvolver um protocolo padrão, eficiente e xeno-livre, para a reprogramação de células-tronco pluripotentes induzidas, que possa ser usado durante as terapias de células-tronco pluripotentes induzidas para o tratamento de doenças degenerativas da retina, e comparar a eficácia relativa de sistemas de cultivo de origem animal e de origem não animal na geração de células-tronco pluripotentes induzidas. Métodos: Cultivos primários de fibroblastos de pterígio humano e de fibroblastos da cápsula de Tenon humanos foram induzidos a células-tronco pluripotentes induzidas usando um vírus não integrado sob dois sistemas xeno-livres; um sistema tradicional de reprogramação não xeno-livre também foi avaliado como parte deste estudo. Os clones de células-tronco pluripotentes induzidas foram selecionados e contados por coloração de células vivas. As eficiências de reprogramação foram avaliadas entre os diferentes fibroblastos e entre os diferentes sistemas de cultivo. Uma série de experimentos, como o PCR e a coloração por imunofluorescência, foram conduzidos para caracterizar as células-tronco pluripotentes induzidas. Resultados: Célu­las-tronco pluripotentes induzidas derivadas de fibroblastos de pterígio humano e fibroblastos da cápsula de Tenon humanos foram estabelecidas com sucesso sob diferentes sistemas de reprogramação e exibiram propriedades de células-tronco pluripotentes induzidas. As eficiências de reprogramação das células-tronco pluripotentes induzidas usando o sistema de terapia celular, o sistema tradicional e o sistema E6/E8 foram 0,014, 0,028% e 0,001%, respectivamente. Além disso, as efi­ciências de reprogramação de células-tronco pluripotentes induzidas derivadas de fibroblastos de pterígio humano e de fibroblastos da cápsula de Tenon humanos usando todos os sistemas acima foram de 0,018% e 0,017%, respectivamente. Conclusões: O vírus Sendai pode ser usado para facilitar a reprogramação de fibroblastos oculares pelas células-tronco pluripotentes induzidas. Tanto os fibroblastos de pterígio humano quanto os fibroblastos da cápsula de Tenon humanos são seguros e eficientes para a reprogramação de células-tronco pluripotentes induzidas. Embora as eficiências de reprogramação das células-tronco pluripotentes induzidas de origem ocular sob condições xeno-livres não tenham sido superiores às eficiências observadas para o sistema tradicional de reprogramação, o sistema de reprogramação sistema de terapia celular é uma boa opção para a indução de células-tronco pluripotentes induzidas sob condições xeno-livres.

Inactivated Sendai Virus Induces ROS-dependent Apoptosis and Autophagy in Human Prostate Cancer Cells / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>The current study aims to investigate the effect of Hemagglutinating virus of Japan envelope (HVJ-E) on induction of apoptosis and autophagy in human prostate cancer PC3 cells, and the underlying mechanisms.</p><p><b>METHODS</b>PC3 cells were treated with HVJ-E at various multiplicity of infection (MOI), and the generated reactive oxygen species (ROS), cell viability, apoptosis, and autophagy were detected, respectively. Next, the role of ROS played in the regulation of HVJ-E-induced apoptosis and autuphagy in PC3 cells were analysed. In the end, the relationship between HVJ-E-induced apoptosis and autuophagy was investigated by using rapamycin and chloroquine.</p><p><b>RESULTS</b>Flow cytometry assay revealed that HVJ-E treatment induced dose-dependent apoptosis and that the JNK and p38 MAPK signaling pathways were involved in HVJ-E-induced apoptosis in PC3 cells. In addition, HVJ-E was able to induce autophagy in PC3 cells via the class III PI3K/beclin-1 pathway. The data also implyed that HVJ-E-triggered autophagy and apoptosis were ROS dependent. When ROS was blocked with N-acetylcysteine (NAC), HVJ-E-induced LC3-II conversion and apoptosis were reversed. Interestingly, HVJ-E-induced apoptosis was significantly increased by an inducer of autophagy, rapamycin pretreatment, both in vitro and in vivo.</p><p><b>CONCLUSION</b>HVJ-E exerts anticancer effects via autophagic cell death in prostate cancer cells.</p>

Reprogramming effects of two kinds of human odontogenic induced pluripotent stem cells and comparison of biological characteristics / 重庆医学

Objective To compare the reprogramming effects and biological characteristics of two types of human odontogen-ic induced pluripotent stem cells(iPSCs).Methods Human dental pulp stem cells(DPSCs)and stem cells from apical papilla (SCAP)were isolated and primarily cultured.The Sendai reprogramming system was utilized to induce DPSCs and SCAP into iP-SCs.The morphology,reprogramming efficiency,reprogramming and time were compared between human DPSCs-iPSCs and SCAP-iPSCs.The SeV and exogenous transcriptional gene expression were detected by RT-PCR.Results Human DPSCs and SCAP were reprogrammed as iPSCs with classical ES-like clonal morphology.The reprogramming efficiencies were(0.68 ± 0.02)% and(0.7 ± 0.01)% respectively,the difference was not statistically significant(P>0.05).The reprogramming time was(26.0 ± 2.1)d and (27.0 ± 1.4)d respectively,the difference was not statistically significant(P>0.05).The RT-PCR results showed that no expres-sion of exogenous virus or transcriptional gene sequence in both iPSCs.Conclusion Human DPSCs and SCAP can be reprogrammed as virus-free and transgene-free iPSCs,which are the ideal sources of iPSCs.

Inactivated Sendai virus induces apoptosis in cisplatin-resistant human lung adenocarcinoma A549 cells in vitro and in vivo / 中国比较医学杂志

Objective Cisplatin(DDP)is widely used in the chemotherapy of lung cancer. However, cisplatin resistance represents a major obstacle in its effective treatment. Our preliminary work has demonstrated that inactivated Sendai virus(HVJ-E)shows that it induces apoptosis in murine melanoma cells(B16)and obviously inhibites the tumor growth in tumor-bearing BALB/c nude mice. This study aims to investigate whether inactivated HVJ-E has an effect of inducing apoptosis in cisplatin-resistant A549/DDP lung adenocarcinoma cells in vitro and in vivo. Methods HVJ-E and A549/DDP cells were co-cultured in vitro,and the effect of HVJ-E on the apoptosis in A549/DDP cells was detected by flow cytometry. In addition,HVJ-E was injected into the tumor in vivo, and its oncolytic effect was observed by TUNEL assay of tissue sections and measurement of tumor size. Results After co-cultured with HVJ-E for 12 h,24 h and 36 h, the apoptosis rate of A549/DDP cells in late stage detected by flow cytometry was 7.7%, 12.6% and 18.9%,respectively,showing a significant difference between 12 h and 24 h, and between 24 h and 36 h. TUNEL assay showed that there was more apoptosis in tumor cells in vivo in the experimental group than in the control group. Meanwhile, intratumoral injection of HVJ-E induced a significantly smaller tumor volume in the experimental group compared with the control group(P ﹤ 0.05). Conclusions Our findings indicate that inactivated HVJ-E can induce apoptosis in A549/DDP cells both in vitro and in vivo, and intratumoral injection of inactivated Sendai virus significantly reduces the tumor growth in vivo.

Reprogramming human dermal fibroblast into induced pluripotent stem cells using non-integrative Sendai virus for transduction

@#Introduction: Induced pluripotent stem cells (iPSC) that exhibit embryonic stem cell-like properties with unlimited self-renewal and multilineage differentiation properties, are a potential cell source in regenerative medicine and cell-based therapy. Although retroviral and lentiviral transduction methods to generate iPSC are well established, the risk of mutagenesis limits the use of these products for therapeutic applications. Materials and Methods: In this study, reprogramming of human dermal fibroblasts (NHDF) into iPSC was carried out using non-integrative Sendai virus for transduction. The iPSC clones were characterised based on the morphological changes, gene expression of pluripotency markers, and spontaneous and directed differentiation abilities into cells of different germ layers. Results: On day 18-25 post-transduction, colonies with embryonic stem cell-like morphology were obtained. The iPSC generated were free of Sendai genome and transgene after passage 10, as confirmed by RT-PCR. NHDF-derived iPSC expressed multiple pluripotency markers in qRT-PCR and immunofluorescence staining. When cultured in suspension for 8 days, iPSC successfully formed embryoid body-like spheres. NHDF-derived iPSC also demonstrated the ability to undergo directed differentiation into ectoderm and endoderm. Conclusion: NHDF were successfully reprogrammed into iPSC using non-integrating Sendai virus for transduction.

A comparative study of two reprogramming systems for inducing pluripotent stem cells from human dental origin / 重庆医学

Objective To comparatively study the features of two reprogramming systems of induced pluripotent stem cells (iPSCs)from human dental origin.Methods Two kinds of reprogramming system,i.e.STEMCCA lentivirus /feed layer and Sen-dai virus /matrigel were used to induce human stem cells from apical papilla(SCAP)into iPSCs,respectively.The induction efficien-cies,workload of generating iPSCs,aneuploidy karyotype ratio,complexities of eliminating exogenous transcription factors and spe-cific markers expression were compared between these two systems.Results The STEMCCA reprogramming system required to prepare the feeder cell MEF.The reprogramming efficiency was 0.1%.Transcription gene-free iPSCs cells were obtained by the Cre-loxp enzyme digestion technique at the later stage.Sendai virus reprogramming system was feeder-free and the preparation of matrigel was quite simple with unified standard.The reprogramming efficiency was 0.7%,which was much higher than that of STEMCCA system(P <0.05).The exogenous virus and transgenes could be gradually eliminated after several passages of natural subclone.Conclusion The Sendai virus/matrigle reprogramming system is much more applicable for the induction of iPSCs from dental origin than the STEMCCA system.

Generation of pluripotent stem cells using non-integrated approach in human azoospermia patients / 天津医药

Objective To establish induced pluripotent stem cells (iPSCs) in patients with azoospermia by non-integrated approach. Methods Using the commercially available serum-free medium (TeSR?2) and embryonic stem cell culture medium (Stem Adhere? Defined Matrix) to define the culture system, the iPSCs were established by using non-integrated Sendai virus infection in peripheral blood mononuclear cells of azoospermia patients. The immunofluorescence, karyotype analysis, embryoid body differentiation and teratoma formation were used to identify pluripotency, karyotype and differentiation ability of iPSCs. Results The established iPSCs showed the characteristics of human embryonic stem cells. Immunofluorescence analysis showed that octamer-binding transcription factor 4 (OCT4), SRY-related-box protein-2 (SOX2), stage-specific embryonic antigen-4 (SSEA-4) and tumor rejection antigen-1-60 (TRA-1-60) were positive for the expression of stem cell pluripotency markers. Karyotype analysis showed that they had normal karyotype. In addition, embryoid body and teratoma tests showed that the iPSCs had the ability to differentiate into three germ layers in vitro and in vivo. Conclusion The induction of pluripotent stem cell line is successfully constructed by non-integrated approach in azoospermia patients.

Establishment of a visualized detection method of Sendai virus by reverse transcription loop-mediated isothermal amplification / 中国实验动物学报

Objective To establish a simple and sensitive detection method of Sendai virus ( SeV ) by reverse transcription loop-mediated isothermal amplification ( RT-LAMP) technique. Methods According to the published Gen-Bank sequences (DQ219803. 1), six pairs of primers were designed targeting the conserved region of SeV. The amplifica-tion products were detected with a LAMP real-time Turbidimeter. (LA-302). Through optimizing the LAMP primers and re-action conditions, a rapid and specific detection method of SeV was established. Meanwhile, the amplified products were colored by fluorescence detection reagent after completion of the reaction, so that the amplification could be visualized and detected by naked eyes. Then, methodological evaluation of the RT-LAMP was tested. Results The method of RT-LAMP showed a highly efficient amplification for SeV viral target gene which was performed at 63℃ for 60 min with the LAMP re-al-time Turbidimeter (LA-302). The detection limit was 2. 1 TCID50, 100 times higher than that of RT-PCR, and no cross-reaction with other RNA and DNA viruses of mice was observed. The results of SeV LAMP reaction was visualized and the tube could be directly observed by naked eyes with the addition of fluorescence detection reagent. The results were consist-ent with the results detected by real-time tubidimeter. 92 clinical samples were detected byRT- LAMP, RT-PCR and indi- rect ELISA, and the coincidence rate was 100%. Conclusions This established SeV RT-LAMP detection method is fast, specific, highly sensitive,easy to perform under simple conditions, and is suitable for rapid detection of Sendai viirus.

Inactivated Sendai virus induces apoptosis mediated by reactive oxygen species in murine melanoma cells / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>This paper aims to investigate the apoptotic effect of inactivated Sendai virus (hemagglutinating virus of Japan-enveloped, HVJ-E) on murine melanoma cells (B16F10) and the possible mechanisms involved in the putative apoptotic reactions.</p><p><b>METHODS</b>B16F10 cells were treated with HVJ-E at various multiplicities of infection (MOI), and the reactive oxygen species (ROS), cell viability, and apoptosis were measured. Next, the roles of ROS in the regulation of Bcl-2/Bax and the activation of mitogen-activated protein kinase (MAPK) pathways in HVJ-E-treated B16F10 cells were analyzed. To further evaluate the cytotoxic effect of HVJ-E-generated ROS on B16F10 cells, HVJ-E was intratumorally injected, both with and without N-acetyl-L-cysteine (NAC), into melanoma tumors on BALB/c mice. Tumor volume was then monitored for 3 weeks, and the tumor proteins were separated for immunoblot assay.</p><p><b>RESULTS</b>Treatment of B16F10 cells with HVJ-E resulted in a dose-dependent inhibition of cell-viability and an induction of apoptosis. The latter effect was associated with the generation of ROS. Inhibition of ROS generation by NAC resulted in a significant reduction of HVJ-E-induced Erk1/2, JNK, and p38 MAPK activation. Additionally, ROS inhibition caused a decrease in the Bcl-2/Bax ratio as well as promoting activation of apoptosis both in vitro and in vivo.</p><p><b>CONCLUSION</b>These results suggest that HVJ-E possesses potential anticancer activity in B16F10 cells through ROS-mediated mitochondrial dysfunction involving the MAPK pathway.</p>

Development of an amplified luminescent proximity homogeneous assay for detecting Sendai virus / 中国比较医学杂志

Objective To establish the amplified luminescent proximity homogeneous assay(AlphaLISA) for the detection of Sendai virus.Methods The antigen concentration,serum concentration and the donor beads/acceptor beads ratio used in the AlphaLISA method were optimumized by the phalanx experiments, then the antibodies of Sendai Virus in 40 rat sera were detected by the established AlphaLISA method and ELISA detection method.The results were compared and the differentia between the two methods was confirmed by IFA.Results The optimum concentration of SV bio-peptide in AlphaLISA assay was 250 nmol/L, the best proportion of donor beads/acceptor beads ratio was 1 ∶1, using the concentration of 20 μg/mL and the serum dilution was 1∶10000.7 of the 40 rat sera were detected SV positive by ELISA, the positive rate was 17.5%, 8 of the 40 rat sera were determined SV positive by AlphaLISA, and the positive rate was 20.0%, the AlphaLISA positive serum was confirmed by IFA.Conclusions We preliminary established the Amplified Luminescent Proximity Homogeneous Assay(AlphaLISA) for the detection of Sendai virus.The sensitivity of the method is comparable to classical ELISA method, but this method use less serum samples and without washing steps.The method has some advantages in degeneracy and accuracy.

Induction of apoptosis in hormone-resistant human prostate cancer PC3 cells by inactivated Sendai virus / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Inactivated Sendai virus particle [hemagglutinating virus of Japan envelope (HVJ-E)] has a potential oncolytic effect due to its ability to induce apoptosis in tumor cells. However, the molecular mechanism of apoptosis induction in cancer cells mediated by HVJ-E has not been fully elucidated. This paper aims to investigate the underlying mechanism of apoptosis induction by HVJ-E in prostate cancer cells (PC3).</p><p><b>METHODS</b>PC3 cells were treated with HVJ-E at various MOI, and then interferon-β (IFN-β) production, and the cell viability and apoptosis were detected by ELISA, MTT-based assay and flow cytometry, respectively. Next, the roles of Jak-Stat, MAPK and Akt pathways played in HVJ-E-induced apoptosis in PC3 cells were analyzed by immunoblot assay. To further evaluate the cytotoxic effect of HVJ-E on PC3 cells, HVJ-E was intratumorally injected into prostate cancers on BALB/c-nude mice, and the tumor volume was monitored for 36 days.</p><p><b>RESULTS</b>HVJ-E induced IFN-β production and activated Jak-Stat signaling pathway, which resulted in the activation of caspase-8, caspase-3, and PARP in PC3 prostate cancer cells post HVJ-E treatment. Furthermore, we observed for the first time that p38 and Jnk MAPKs in PC3 cells contributed to HVJ-E-induced apoptosis. In addition, intratumoral HVJ-E treatment displayed a direct inhibitory effect in an in vivo BALB/c nude mouse prostate cancer model.</p><p><b>CONCLUSION</b>Our findings have provided novel insights into the underlying mechanisms by which HVJ-E induces apoptosis in tumor cells.</p>

Study on the apoptosis of rat glioma cells induced by defective interfering particles of Sendai virus strain Tianjin / 中华微生物学和免疫学杂志

Objective To investigate the apoptosis of rat glioma C 6 cells induced by defective in-terfering( DI) particles of Sendai virus strain Tianjin .Methods Rat glioma C6 cells were treated with dif-ferent titers of DI particles of Sendai virus strain Tianjin in vitro with culture media as negative control and intact virus as positive control .At different time point , cells were collected and their apoptosis was detected by DNA gel electrophorsis , TUNEL assay and AnnexinⅤ/PI double-labeled flow cytometry .The C6 glioma-bearing rat model was established and then treated with three intratumoral injections of DI particles , intact virus or saline three times at interval of two days .The antitumor effects of ID particles were evaluated through daily measuring of the tumor size .Hematoxylin-eosin( HE) staining was used to observe the patho-logical changes in tumor tissues .TUNEL assay was performed to detect the apoptosis of tumor tissues .Re-sults Rat glioma C6 cells treated with DI particles or intact virus in vitro showed typical DNA ladder pattern in agarose gel electrophoresis in a time-and dose-dependent manner .With the intervention of DI particles , the apoptosis rate of C6 cells showed a time-and dose-dependent manner and was significantly higher than that of the control group (P0.05).Conclusion The DI particles of Sendai virus strain Tianjin could induce apoptosis of rat glioma C 6 cells in a time-and dose-dependent manner both in vitro and in vivo, suggesting that the DI particles might be applicable for the treatment of neurogliocytoma in the future.

Microbiological Contamination of Laboratory Mice and Rats in Korea from 2007 to 2008 / 한국실험동물학회지

In order to assess the microbiological contamination of laboratory mice and rats in Korea over the 2-year period from 2007 to 2008, we monitored animals housed in mouse and rat facilities equipped with barrier systems. In a barrier animal facility in Korea, the most important viruses in the identified pathogen were the mouse hepatitis virus (MHV) and Pasteurella (Pa.) pneumotropica, and Staphylococcus aureus was identified as the most common bacterial pathogen in Korea. The most commonly detected parasite in the identified pathogen was Trichomonas spp. in the mouse facilities and Entamoeba spp. in the rat facilities. In many cases, these pathogen-contaminated animals were genetically modified animals obtained from the university. Currently, consistent with the increased transfer of genetically modified animals between domestic and foreign animal facilities, the Pa. pneumotropica and parasites infection rates were shown to have increased as compared to those of the 2004-2006 period. Indeed, the MHV infection rate has been maintained at almost 20% in Korean animal facilities over the past 10 years. These results showed that effective quarantine programs for contaminated genetically engineered mutant mice and the monitoring of regular or irregular MHV monitoring in animal colonies should help to reduce pathogen contamination in Korean animal facilities.

Microbiological Contamination of Laboratory Mice and Rats in Korea from 2007 to 2008 / 한국실험동물학회지

In order to assess the microbiological contamination of laboratory mice and rats in Korea over the 2-year period from 2007 to 2008, we monitored animals housed in mouse and rat facilities equipped with barrier systems. In a barrier animal facility in Korea, the most important viruses in the identified pathogen were the mouse hepatitis virus (MHV) and Pasteurella (Pa.) pneumotropica, and Staphylococcus aureus was identified as the most common bacterial pathogen in Korea. The most commonly detected parasite in the identified pathogen was Trichomonas spp. in the mouse facilities and Entamoeba spp. in the rat facilities. In many cases, these pathogen-contaminated animals were genetically modified animals obtained from the university. Currently, consistent with the increased transfer of genetically modified animals between domestic and foreign animal facilities, the Pa. pneumotropica and parasites infection rates were shown to have increased as compared to those of the 2004-2006 period. Indeed, the MHV infection rate has been maintained at almost 20% in Korean animal facilities over the past 10 years. These results showed that effective quarantine programs for contaminated genetically engineered mutant mice and the monitoring of regular or irregular MHV monitoring in animal colonies should help to reduce pathogen contamination in Korean animal facilities.

Bioinformatics Analysis of the NP, P, M and L Protein of Paramyxovirus Tianjin Strain / 微生物学通报

In order to demonstrate the taxonomic position of paramyxovirus Tianjin strain and explore its mechanism of pathogenesis. Bioinformatics methods were used to analyze the deduced amino acid sequences of NP, P, M, and L protein of Tianjin strain. Phylogenetic analysis based on NP, P, M, and L protein sequences demonstrated that Tianjin strain belonged to the genus Respirovirus, in the subfamily Paramyxovirinae and most likely a new genotype of Sendai virus. Sequence similarities comparisons indicated that Tianjin strain P protein was poorly conserved, sharing only 78.7%~91.9% amino acid identity with 6 known Sendai viruses, while L protein was the most conserved, having 96.0%~98.0% amino acid identity with other Sendai viruses. Multiple-sequence alignments of Tianjin strain NP, P, M, and L protein with those of 6 known Sendai viruses showed that Tianjin strain possessed a lot of unique amino acid substitutions in protein sequences, 15 in NP, 29 in P, 6 in M, and 29 in L. The presence of these unique amino acid substitutions suggests that Tianjin strain maybe has a significant difference in host or pathological characteristics from the known Sendai viruses.

Ca2+ and the interaction of pore-formers with membranes.

The interaction of pore-forming agents, such as Sendai virus, influenza virus (at pH 5 3), activated complement, Staphylococcus aureus α-toxin, melittin and polylysine, with the surface membrane of cells has been studied. In each case the following changes are initiated: collapse of membrane potential, leakage of ions, and leakage of phosphorylated metabolites. The changes can be inihibited by extracellular Ca2+ at physiological concentration; Mg2+ is less effective, and Zn2+ is more effective, than Ca2+ Ca2+ appears to act at a stage subsequent to the binding of pore-forming agent to cells. It is concluded that divalent cations are able to protect cells against the damaging effects of certain viruses, toxins or the components of activated complement in a manner that is worthy of further investigation.

Effect of sendai virus replication sequence on protein changes in immature dendritic cells / 第三军医大学学报

Objective To investigate the protein change in immature dendritic cells(DC) exposed to sendai virus replication sequence.Methods The total cellular proteins expressed in DC infected with Sev/△F or SeV were collected and quantitated,then separated by two-dimensional gel electrophoresis and visualized by silver staining.Digital images were analyzed by PDQuest software.The differentially expressed protein spots were picked,digested in gel and subjected to peptide mass fingerprinting using MALDI-TOF MS.Results Protein spots in 2-DE gel for DC treated by Sev/△F were more than that by SeV,among which a lot of proteins increased.Conclusion It was relevant to the replication sequence that proteins decreased in dendritic cells infected with sendai virus vector.