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.     

人胚雪旺细胞组织工程神经修复坐骨神经缺损的实验研究

目的：探索人胚雪旺细胞作为组织工程的种子细胞修复周围神经缺损的可行性。方法：通过组织工程方法用PLGA导管和polyglactin 910纤维负载人胚雪旺细胞预先构置好人工神经，然后用于修复大鼠20mm的坐骨神经缺损，并与神经切断后原位缝合以及用单纯的PLGA导管进行修复的实验组进行对照。通过活体肢体功能观察、靶器官肌肉测量、电生理检测、辣根过氧化物酶示踪、连续组织切片图像分析以及透射电镜等检查神经再生情况。结果：人工神经修复组神经再生良好，效果接近于神经原位缝合组，明显优于单纯的PLGA导管修复组。结论：人胚雪旺细胞构建的人工神经可以修复20mm的周围神经缺损。     

趾蹼血管分型的临床认识

目的分析120例足趾移植再造手指病例，对趾蹼间的血管形态和交通情况进行观察，并阐述其临床意义。方法临床观察120例156侧趾蹼，对第一跖骨背动脉和第一趾底动脉在趾蹼间的走向、口径以及两者的交通支情况进行记录并进行分型。结果按孙博的分型方法：Ⅰ型72侧，占46．2％；Ⅱ型57侧，占36．5％；Ⅱ型7侧，占4．5％；Ⅳ型3侧，占1．9％；Ⅴ型15侧，占9．6％；另有2侧（占1．3％）第一跖骨背血管口径〉1mm，虽发出趾背动脉供养蹲趾胫侧和第二趾腓侧，但在趾蹼内与趾底血管无吻合，故称之为孙博Ⅵ型。结论（1）从解剖形态看应增加孙博Ⅵ型较全面；（2）从临床实用角度出发，趾蹼间血管可以简单分为三型：Ⅰ型交通支口径在0．5mm以上。Ⅱ型交通支在0．5mm以下。Ⅲ型无交通；（3）手术中采用第一套或第二套供血系统，首先要看趾蹼间血管的交通支，即按本组的分型属Ⅰ型采用第一套供血系统，Ⅱ、Ⅲ型采用第二套供血系统。     

树枝状静脉移植在手部创伤中的应用

目的探讨手掌严重碾挫伤应用血管移植手术方法的选择。方法对12例手掌碾挫伤伴血管损伤的患者，采用树枝状浅表静脉倒转后多级移植修复缺损的手掌动脉，同时对缺损皮肤一期行植皮或皮瓣修复。结果术后12例创面全部Ⅰ期愈合，手指血循环好。术后随访3～22个月，手功能评定： 优5例，良4例，可2例，差1例，优良率为75％。手掌恢复了良好的外形和功能，能完成手指的捏、握、对掌和对指功能。结论对严重的手掌毁损伤，利用树枝状静脉移植修复缺损的血管，能获得较好的手功能和外形。     

窒息鼠脑组织型纤溶酶原激活物活性变化与脑水肿的关系

目的：探讨窒息对鼠脑分泌组织型纤溶酶原激活物（TPA）的影响与脑水肿的关系。方法：通过“延迟剖宫产术”致胎鼠宫内窘迫，实验分空白对照组，窒息15min组，窒息30min组，窒息15min复氧30min组，窒息30min复氧30min五个实验组，每组各取8例测试脑组织TAP的活性及含水量，结果：窒息后鼠脑TPA活性与含水量均升高（P＜0．01），结论：窒息可致TAP活性增高，同时发生脑水肿，高活性的TPA可能是脑缺氧缺血致不可逆神经元损伤的一个重要媒介。     

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.     

应用皮瓣移植修复足跟部皮肤软组织缺损

目的通过8种不同皮瓣的临床应用,总结足跟部皮肤软组织缺损的修复效果。方法采用了游离足背皮瓣、游离背阔肌皮瓣、游离肩胛皮瓣、桥式胫后动脉岛状皮瓣、桥式携带背阔肌皮瓣或桥式携带肩胛皮瓣、胫后动脉岛状皮瓣、胫后动脉皮支岛状皮瓣、跖底内侧动脉岛状皮瓣,对足跟部皮肤软组织缺损进行修复的临床应用。结果95块皮瓣修复足跟部皮肤软组织缺损,成活93块,成功率为97.9%。随访1～15年,治疗效果满意。结论应用皮瓣移植的方法,才能使足跟部皮肤软组织缺损得到有效的修复。     

组织工程血管管状支架的制备与动态培养在血管组织工程中的应用

目的探索体外构建组织工程血管及细胞种植的方法。方法(1)制备管状支架模具,采用模具成形,冷冻干燥等技术,应用胶原和透明质酸复合体外预构管状支架;(2)分动态种植和静态培养两组进行细胞分层种植,每组12个,应用水溶性磺酸四唑(WST-1)测定,扫描电镜观察,5-溴2-脱氧嘧啶核苷(BrdU)细胞标记鉴定比较两种方法的细胞生长情况。结果(1)成功制备了管状支架,在培养液中约3周可保持管腔形态不变形,不塌陷;2.WST-1测定值在培养6、14、20 d分别为:动态组0.842±0.110、1.124±0.102、1.132±0.112,静态组0.386±0.121、0.489±0.245、0.499±0.265,两组比较差异有统计学意义,电镜和BrdU标记表明动态组细胞排列整齐有序。结论应用模具成形,冷冻干燥技术预构管状支架的方法切实可行,动态培养有助于细胞种植,本研究为组织工程血管的构建方法提供了思路。     

多发性骨髓瘤患者血浆尿激酶型纤溶酶原激活物及其可溶性受体的检测与临床意义

目的 :研究多发性骨髓瘤 ( MM)患者血浆尿激酶型纤溶酶原激活物 ( u- PA )及其可溶性受体 ( su PAR )的水平变化 ,并探讨其临床意义。方法 :用 ELISA法检测 34例 MM患者血浆 u- PA及 su PA R的浓度 ,同时观察其中6例 MM患者化疗前后血浆 u- PA及 su PAR的浓度变化。结果 :MM患者血浆 u- PA及 su PA R水平均明显高于正常对照组 ,其中进展期 MM患者血浆 u- PA及 su PAR水平明显高于正常对照组和稳定期 MM患者 ( P <0 .0 1) ,而稳定期 MM患者血浆 u- PA及 su PA R水平与正常对照组无显著性差异 ( P>0 .0 5)。 6例 MM患者化疗后血浆 u- PA及 su PA R水平 ,明显低于化疗前血浆 u- PA及 su PAR水平 ( P<0 .0 5)。骨髓涂片瘤细胞比例 >2 0 %的 MM患者血浆 u- PA及 su PA R水平 ,明显高于瘤细胞比例≤ 2 0 % M M患者 ( P<0 .0 5;P<0 .0 1)。M M患者血浆 u- PA及su PA R水平均与骨髓瘤细胞百分比及血清球蛋白呈正相关 ,而与血清白蛋白呈负相关。结论 :血浆 u- PA及 su PA R水平升高可能与多发性骨髓瘤的发生、发展有密切关系 ;其水平可作为临床分期、判断疗效、了解疾病进展情况及预后的一个重要指标。