A DNA vaccine expressing an optimized secreted FAPα induces enhanced anti-tumor activity by altering the tumor microenvironment in a murine model of breast cancer

Cancer-associated fibroblasts (CAFs), major components of the tumor microenvironment (TME), promote tumor growth and metastasis and inhibit the anti-tumor immune response. We previously constructed a DNA vaccine expressing human FAPα, which is highly expressed by CAFs, to target these cells in the TME, and observed limited anti-tumor effects in the 4T1 breast cancer model. When the treatment time was delayed until tumor nodes formed, the anti-tumor effect of the vaccine completely disappeared. In this study, to improve the safety and efficacy, we constructed a new FAPα-targeted vaccine containing only the extracellular domain of human FAPα with a tissue plasminogen activator signal sequence for enhanced antigen secretion and immunogenicity. The number of CAFs was more effectively reduced by CD8⁺ T cells induced by the new vaccine. This resulted in decreases in CCL2 and CXCL12 expression, leading to a significant decrease in the ratio of myeloid-derived suppressor cells in the TME. Moreover, when mice were treated after the establishment of tumors, the vaccine could still delay tumor growth. To facilitate the future application of the vaccine in clinical trials, we further optimized the gene codons and reduced the homology between the vaccine and the original sequence, which may be convenient for evaluating the vaccine distribution in the human body. These results indicated that the new FAPα-targeted vaccine expressing an optimized secreted human FAPα induced enhanced anti-tumor activity by reducing the number of FAPα⁺ CAFs and enhancing the recruitment of effector T cells in the 4T1 tumor model mice.     

Study on the Mechanism of the Annexin I -Mediated Co-Assembly of t-PA and Plasminogen

In order to further investigate the effect of annexin Ⅱ (Ann- Ⅱ ) on tissue plasminogen activator (t-PA)-dependent plasminogen (PLG) activation and its interactive mechanism, recombinant native Ann- Ⅱ bound t-PA, PLG and plasmin with high affinity was examined. The flow cytometric assay showed that the ann- Ⅱ expression rate was higher in the human umbilical vein endothelial cell (HUVEC) (87. 65 %) than in the HL-60 cells as controls (35. 79 %). Two irrelevant proteins,bovine serum albumin (BSA) and equine IgG (EIG) had no effect on the production of plasmin.Ann- Ⅱ -mediated enhancement of t-PA-dependent PLG activation was inhibited by ε-aminocaproic acid or by pretreatment of Ann- Ⅱ with carboxypeptidase B with the inhibitive rate being 77.8 % and 77. 0 %, respectively. It was revealed that the effect of Ann- Ⅱ on PLG activation was specific for tPA. Urokinase didn‘t bind to Ann- Ⅱ , demonstrating the role of receptor-related lysine residues on activation of PLG, showing that the Ann- Ⅱ -PLG interaction was dependent upon carboxyl-terminal lysine residues. These findings suggest that annexin Ⅱ -mediated co-assembly of t-PA and PLG may promote plasmin generation and play a key role in modulating fibrinolysis on the endothelial surface.     

年龄与代谢综合征的关系及防治对策

目的了解中老年人群年龄与多种代谢异常的聚集状况,为心脑血管疾病的一级预防提供依据。方法对744例中老年人按不同年龄分为5组,分析代谢综合征(MS)各项指标的变化规律及特点。结果年龄与肥胖呈明显负相关(P<0.001),与高血糖、高血压及冠心病、脑卒中呈明显正相关(P<0.05或0.001);40～60岁组MS的患病率基本一致,约为30%,70岁以后明显增加,到80岁时可高达51%(P<0.05或0.01);MS与冠心病和脑卒中的患病率均明显相关(P<0.05),随着临床指标个数的增加,冠心病的患病率增加了6.8%,脑卒中的患病率增加了7.44%。结论中年人群超重或肥胖的患病率明显高于老年组,应引起足够重视;随着年龄增长,代谢异常指标数目增多,尤其是≥3项指标的人数明显增加;MS作为心血管危险因素直接导致冠心病、脑卒中发病率增加。针对中老年人群MS的特点,制定相应的干预措施十分重要。     

Changes in Neutrophil Oxidative Potential in Patients Undergoing Cardiopulmonary Bypass with Polypropylene Hollow Fiber Oxygenators

Neutrophil oxidative metabolism, C3d and beta 2 microglobulin levels, were assessed in nine consecutive patients undergoing cardiopulmonary bypass surgery with polypropylene hollow fiber oxygenators for open cardiac operations. Generation of oxygen free radicals by neutrophils was measured as luminol-enhanced chemiluminescence after stimulation with opsonized Zymosan and phorbol myristate acetate. A significant increase in light emission was detected by using both of the chemiluminescence stimulators. Moreover, a remarkable and significant increase in C3d levels was found already at 10 min. Conversely minimal changes in levels of beta 2 microglobulin were detected during cardiopulmonary bypass surgery. These data suggest that the impact of the patient blood with the foreign surface of cardiopulmonary bypass results in activation of phagocyte cells with increased potential in oxygen consumption. These effects could be partially complement-mediated.     

Study on lymphocyte activation and proliferation induced by anti-CD3 McAb

Summary T cell activation and proliferation via CD₃-TCR complex were investigated by lymphocyte DNA synthesis in vitro. Several interfering factors were also discussed. The result indicated that lymphocyte activation and proliferation are calciumdependent. A rise of cytoplasmic free Ca²⁺ quickly following activation with CD₃ McAb is mainly due to intracellular mobilization of Ca²⁺, while lymphocyte proliferation needs both intracellular mobilization of Ca²⁺ as well as influx of extracellular Ca²⁺. It was confirmed that CTX sensitive G protein plays a role in regulating T cell proliferation by pretreatment with CTX suppressing lymphocyte³H-TdR incorporation obviously. PLC and PKC inhibitor neomycin and P. S. S could also decrease T cell proliferation.     

Mutagenicity of benzo(a)pyrenyl-1-sulfate in the Ames test.

Comparison of the mutagenicity of nine isomeric benzo(a)pyrenyl [B(a)P] phenols conjugated with either sulfate or glucuronide was carried out using strain Salmonella typhimurium TA98. Of the nine conjugates tested, only B(a)P-1-sulfate was mutagenic. Accordingly, the mutagenicity of B(a)P-1-sulfate was compared with that of B(a)P and 1-hydroxybenzo(a)pyrene [B(a)P-1-OH] in the presence and absence of rat lung S9 and Aroclor-induced liver S9 with and without an NADPH-generating system. B(a)P-1-sulfate was slightly mutagenic, whereas B(a)P and the 1-hydroxy derivative were nonmutagenic when S9 fractions and NADPH were omitted. Addition of induced liver S9 with NADPH caused mutagenicity with B(a) -1-OH greater than B(a)P greater than B(a)P-1-sulfate. B(a)P-1-sulfate was the only mutagenic species when lung S9 was added. This mutagenicity did not require NADPH. Sodium sulfite, an inhibitor of arylsulfatase, decreased the mutagenicity of B(a)P-1-sulfate. These data suggest that a unique mutagenic species is generated from B(a)P-1-sulfate via arylsulfatase in rat lung.