枣庄市2008年麻疹减毒活疫苗强化免疫效果评价

为了评价枣庄市麻疹疫苗(MV)强化免疫的效果,于2008年在所辖区开展了流行病学调查、麻疹血清学监测和费用-效益分析等,对MV强化免疫进行了评价。枣庄市麻疹年平均报告发病率在强化免疫前为14.1/10万,对1-6岁儿童开展MV强化免疫后,2010年麻疹发病率为0.8/10万,降低了93.5%;1-6岁儿童麻疹发病构成,强化免疫前为39.87%,强化免疫后为25.90%,大大减少了5岁以下儿童的麻疹发病;强化免疫后,1-6岁儿童麻疹抗体阳性率为99.3%,抗体几何平均滴度(GMT)从强化免疫前的1∶563.0增长到强化免疫后的1∶814.9。开展MV强化免疫费用-效益比值为1∶2.23~1∶3.12。开展MV初始化强化免疫是加速麻疹控制最有效的措施之一。     

Characterizing the role of mechanical signals in gene regulatory networks using Long SAGE

A systems understanding of mechanical regulation is critical for determining how cells proliferate and differentiate. To better understand the biological process in which mechanical signals regulate cells, we globally investigated the gene expression profiling via long serial analysis of gene expression (Long SAGE) in osteoblasts after exposure to mechanical stretching. The analysis showed that the differentially expressed genes were related with many physiological processes, including signal transduction, cell proliferation and apoptosis. Several genes that were seldom or never studied in osteoblasts have been found in this study. We further analyzed the signal pathways and provided gene regulatory networks activated by mechanical signals. Many changed genes in our data were contributed to ECM-integrin-FAK mediated pathway and mainly influenced actin-cytoskeleton dynamic remodeling, cell proliferation and differentiation. We also provided evidence supporting the hypothesis that endoplasmic reticulum and mitochondrion were combined to dedicate to calcium regulation. Taken together, our experiments provided a systemic view on biological processes and mechanotransduction network in osteoblasts, suggesting that mechanical signals regulate osteoblast through a greater diversity of interactions and pathways than previously appreciated.     

A proteoliposome containing apolipoprotein A-I mutant (V156K) enhances rapid tumor regression activity of human origin oncolytic adenovirus in tumor-bearing zebrafish and mice

We recently reported that the efficiency of adenoviral gene delivery and virus stability are significantly enhanced when a proteoliposome (PL) containing apolipoprotein (apo) A-I is used in an animal model. In the current study, we tested tumor removal activity of oncolytic adenovirus (Ad) using PL-containing wildtype (WT) or V156K. Oncolytic Ad with or without PL was injected into tumors of zebrafish and nude mice as a Hep3B tumor xenograft model. The V156K-PL-Ad-injected zebrafish, group showed the lowest tumor tissue volume and nucleic acids in the tumor area, whereas injection of Ad alone did not result in adequate removal of tumor activity. Reactive oxygen species (ROS) contents increased two-fold in tumor-bearing zebrafish; however, the V156K-PL-Ad injected group showed a 40% decrease in ROS levels compared to that in normal zebrafish. After reducing the tumor volume with the V156K-PL-Ad injection, the swimming pattern of the zebrafish changed to be more active and energetic. The oncolytic effect of PL-Ad containing either V156K or WT was about two-fold more enhanced in mice than that of Ad alone 34 days after the injection. Immunohistochemical analysis revealed that the PL-Ad-injected groups showed enhanced efficiency of viral delivery with elevated Ad-E1A staining and a diminished number of proliferating tumor cells. Thus, the antitumor effect of oncolytic Ad was strongly enhanced by a PL-containing apoA-I and its mutant (V156K) without causing side effects in mice and zebrafish models.     

Distinct actions of strontium on mineral formation in matrix vesicles

Matrix vesicles (MVs) are involved in the initial step of mineralization in skeletal tissues and provide an easily model to analyze the hydroxyapatite (HA) formation. Sr stimulates bone formation and its effect was tested on MVs. Sr²⁺ (15-50 μM) in the mineralization medium containing MVs, 2 mM Ca²⁺ and 3.42 mM P_i, retarded HA formation. Sr²⁺ (1-5 mM) in the same medium-induced other types of mineral than HA and cancelled the ATP-, ADP- or PP_i-induced retardation in the mineral formation. Our findings suggest that the beneficial effect of Sr²⁺ at a low dose (15-50 μM) is rather an inhibitor of bone resorption than an activator of mineral formation, while at high Sr²⁺ concentration (1-5 mM), mineral formation, especially other types of mineral than HA, is favored.     

靶向Wnt信号的溶瘤腺病毒抑制肝癌干细胞研究

肝细胞癌(hepatocellular carcinoma,HCC)是全球第五大癌症并成为癌症死亡的主要原因,传统治疗早期肝癌取得了一定的进展,但是癌症的复发、转移和耐药仍未得到根本解决,这些现象可通过癌症干细胞理论(cancer stem cell,CSC)进行解释.本研究通过悬浮富集培养的方法,获得了MHCC-97H细胞的三维立体球细胞(sphere cell),并检测其干细胞特性,通过删除5型腺病毒的E1A CR2区域24 bp碱基,并用Wnt活性转录元件TCF/TEF调控E1A基因表达,同时插入抗癌基因TSLC1,得到了双靶向溶瘤腺病毒Ad.wnt-E1A(△24 bp)-TSLC1,通过MTT、结晶紫染色实验、Hoechst、细胞划痕、蛋白质印迹技术、Transwell及免疫荧光技术检测重组病毒对肝癌类干细胞的EMT(epithelial-mesenchymal transition)转化、杀伤、凋亡以及迁移的作用.结果表明,悬浮富集培养的MHCC-97H sphere细胞具有自我更新、分化能力、静息性以及耐药性,高表达肝癌干细胞表面标志物(如CD133等),重组病毒处理后表现出明显的杀伤效果及抑制细胞迁移与侵袭的特性,靶向抑制MHCC-97H sphere细胞能力更强(P0.001),且重组病毒能有效诱导肝癌类干细胞通过caspase途径发生凋亡.因此,重组病毒Ad.wnt-E1A(△24 bp)-TSLC1有可能成为靶向肝癌干细胞的治疗药物,具有一定的临床应用前景.     

The viral approach to breast cancer immunotherapy

Despite years of intensive research, breast cancer remains the leading cause of death in women worldwide. New technologies including oncolytic virus therapies, virus, and phage display are among the most powerful and advanced methods that have emerged in recent years with potential applications in cancer prevention and treatment. Oncolytic virus therapy is an interesting strategy for cancer treatment. Presently, a number of viruses from different virus families are under laboratory and clinical investigation as oncolytic therapeutics. Oncolytic viruses (OVs) have been shown to be able to induce and initiate a systemic antitumor immune response. The possibility of application of a multimodal therapy using a combination of the OV therapy with immune checkpoint inhibitors and cancer antigen vaccination holds a great promise in the future of cancer immunotherapy. Display of immunologic peptides on bacterial viruses (bacteriophages) is also increasingly being considered as a new and strong cancer vaccine delivery strategy. In phage display immunotherapy, a peptide or protein antigen is presented by genetic fusions to the phage coat proteins, and the phage construct formulation acts as a protective or preventive vaccine against cancer. In our laboratory, we have recently tested a few peptides (E75, AE37, and GP2) derived from HER2/neu proto-oncogene as vaccine delivery modalities for the treatment of TUBO breast cancer xenograft tumors of BALB/c mice. Here, in this paper, we discuss the latest advancements in the applications of OVs and bacterial viruses display systems as new and advanced modalities in cancer immune therapeutics.     

Kinetics of the plastoquinone pool oxidation following illumination Oxygen incorporation into photosynthetic electron transport chain

The oxidation of the PQ-pool after illumination with 50 or 500 micromol quantam(-2)s(-1) was measured in isolated thylakoids as the increase in DeltaA(263), i.e., as the appearance of PQ. While it was not observed under anaerobic conditions, under aerobic conditions it was biphasic. The first faster phase constituted 26% or 44% of total reappearance of PQ, after weak or strong light respectively. The dependence on oxygen presence as well as the correlation with the rate of oxygen consumption led to conclusion that this phase represents the appearance of PQ from PQ(*-) produced in the course of PQH(2) oxidation by superoxide accumulated in the light within the membrane.     

Impairment of the photosynthetic apparatus by oxidative stress induced by photosensitization reaction of protoporphyrin IX

Treatment with the herbicide acifluorfen-sodium (AF-Na), an inhibitor of protoporphyrinogen oxidase, caused an accumulation of protoporphyrin IX (Proto IX) , light-induced necrotic spots on the cucumber cotyledon within 12-24 h, and photobleaching after 48-72 h of light exposure. Proto IX-sensitized and singlet oxygen (¹O₂)-mediated oxidative stress caused by AF-Na treatment impaired photosystem I (PSI), photosystem II (PSII) and whole chain electron transport reactions. As compared to controls, the F_v/F_m (variable to maximal chlorophyll a fluorescence) ratio of treated samples was reduced. The PSII electron donor NH₂OH failed to restore the F_v/F_m ratio suggesting that the reduction of F_v/F_m reflects the loss of reaction center functions. This explanation is further supported by the practically near-similar loss of PSI and PSII activities. As revealed from the light saturation curve (rate of oxygen evolution as a function of light intensity), the reduction of PSII activity was both due to the reduction in the quantum yield at limiting light intensities and impairment of light-saturated electron transport. In treated cotyledons both the Q (due to recombination of Q_A⁻ with S₂) and B (due to recombination of Q_B⁻ with S₂/S₃) band of thermoluminescence decreased by 50% suggesting a loss of active PSII reaction centers. In both the control and treated samples, the thermoluminescence yield of B band exhibited a periodicity of 4 suggesting normal functioning of the S states in centers that were still active. The low temperature (77 K) fluorescence emission spectra revealed that the F₆₉₅ band (that originates in CP-47) increased probably due to reduced energy transfer from the CP47 to the reaction center. These demonstrated an overall damage to the PSI and PSII reaction centers by ¹O₂ produced in response to photosensitization reaction of protoporphyrin IX in AF-Na-treated cucumber seedlings.     

Development of a second-generation oncolytic Herpes simplex virus expressing TNFalpha for cancer therapy

BACKGROUND: Tumour necrosis factor alpha (TNFalpha) therapy is a promising anti-cancer treatment when combined with radiotherapy due to its potent radio sensitising effects, but systemic toxicity has limited its clinical use. Previously, non-replicative adenovirus vectors have been used to deliver TNFalpha directly to the tumour, including under the control of a radiation sensitive promoter. Here, we have used an ICP34.5 deleted, oncolytic herpes simplex virus (HSV) for delivery to increase expression levels and spread through the tumour, and the use of the US11 true late HSV promoter to limit expression to where the virus replicates, i.e. selectively in tumour tissue. METHODS: TNFalpha expression under the CMV or US11 promoter was compared on cell lines CT26, BHK and Fadu. To further compare the activities of the promoters, expression of human TNFalpha was analysed in the presence and absence of acyclovir--an inhibitor of viral DNA replication and on HSV/ICP34.5- non-permissive cell line 3T6. The in vivo efficacy and toxicity of TNFalpha viruses were compared using A20 double flank tumour model in Balb/C mice and Fadu tumour model in nude mice. RESULTS: The results demonstrated that the US11 promoter significantly reduced and delayed TNFalpha expression as compared to use of the CMV promoter, especially in non-permissive cells or in the presence of acyclovir. Despite the reduced and more selective expression levels, US11 driven TNFalpha showed improved anti-tumour effects compared to CMV driven TNFalpha, and without the toxic side effects. CONCLUSIONS: This approach is therefore beneficial in increasing localised TNFalpha expression as compared to the use of non-replicative approaches, and combines the effects of TNFalpha with oncolytic virus replication which is expected to further enhance the efficacy of radiotherapy in a combined treatment approach.