转内皮抑制素基因治疗大鼠角膜新生血管

目的:探讨转染内皮抑制素(Endostatin)基因对酸烧伤引起的大鼠角膜新生血管的基因治疗。方法:通过限制性内切酶酶切反应、聚合酶链式反应、DNA测序及用NCBIBLAST软件与基因库序列比较的方法进行鉴定,鉴定后在大肠杆菌中扩增,用质粒纯化试剂盒抽提纯化;用750g/L硝酸银和250g/L硝酸钾混合烧灼液制作大鼠角膜新生血管模型,用结膜下注射脂质体包裹的质粒pBlast- hEndostatin来进行体内基因治疗。结果:实验证实质粒pBlast-hEndostatin含有人endostatin基因。结膜下注射脂质体包裹的质粒Tel押029-82245172Email押IJO.2000@163.compBlast-hEndostatin对酸烧伤引起的炎症性角膜新生血管有明显抑制作用,术后6,10,15d对角膜新生血管面积的抑制率分别为37%,40%,43%;对角膜新生血管密度也有明显的抑制作用,抑制率达40%。对角膜新生血管长度和角膜炎症细胞没有明显抑制作用。角膜新生血管面积与角膜水肿、角膜混浊呈正相关。结论:用结膜下注射脂质体包裹的内皮抑制素基因可以部分抑制酸烧伤引起的大鼠角膜新生血管。其作用机制是转基因产生的内皮抑制素蛋白直接抑制角膜新生血管的形成,而不是通过抑制炎症反应来抑制角膜新生血管的形成。     

Clinical evaluation of ZD6474, an orally active inhibitor of VEGF and EGF receptor signaling, in patients with solid, malignant tumors.

BACKGROUND: ZD6474 selectively inhibits the tyrosine kinase activity of vascular endothelial growth factor receptor and epidermal growth factor receptor. The safety, tolerability and pharmacokinetics of ZD6474 were assessed in a phase I dose-escalation study of patients with advanced solid tumors. PATIENTS AND METHODS: Adult patients with tumors refractory to standard treatments received once-daily oral ZD6474 (50-600 mg) in 28-day cycles, until disease progression or unacceptable toxicity was observed. RESULTS: Seventy-seven patients were treated at doses of 50 mg (n=9), 100 mg (n=19), 200 mg (n=8), 300 mg (n=25), 500 mg (n=8), and 600 mg (n=8). Adverse events were generally mild, and the most common dose-limiting toxicities (DLT) were diarrhea (n=4), hypertension (n=4), and rash (n=3). The incidence of most adverse events appeared to be dose-dependant. In the 500 mg/day cohort, 3/8 patients experienced DLT and this dose was therefore considered to exceed the maximum tolerated dose. Pharmacokinetic analysis confirmed that ZD6474 was suitable for once-daily oral dosing. CONCLUSIONS: Once-daily oral dosing of ZD6474 at 300 mg/day is generally well tolerated in patients with advanced solid tumors, and this dose is being investigated in phase II trials.     

苦参黄酮抑制血管新生活性成分的虚拟筛选

血管新生是一个动态的、多步骤的过程,现在已知大约70种疾病与血管新生紊乱相关。作者前期研究及文献报道均表明苦参黄酮类成分有明显抑制血管新生的作用,但其药效物质基础和作用机制尚未明确。该研究应用分子对接技术虚拟筛选苦参黄酮抑制血管新生的药效物质,搜集现已分离鉴定的126个苦参黄酮类化合物组成配体数据库,选择VEGF-a,TEK,KDR等6个与血管新生密切相关的靶点组成受体数据库,以DrugBank中对各靶点有抑制作用并已上市的小分子药物为参照,设定各靶点对应的已上市小分子药物最低打分为阈值,应用Discovery Studio 2.5(DS2.5)软件的LibDock模块进行分子对接,虚拟筛选出打分高于阈值且排名前10%的化合物共37个。对比分析了原配体、已上市药物和苦参黄酮作用于各靶点的主要活性位点,初步揭示了苦参黄酮抑制血管新生的作用机制,为研发血管新生抑制剂类药物提供了一定的参考。     

Targeted therapies in the treatment of pancreatic carcinoma

Pancreatic carcinoma is the fifth leading cause of cancer deaths in the western world. Although some progress has been made in the clinical management of pancreatic cancer, surgical resection remains the only potentially curative treatment. New strategies are urgently needed for the treatment of this disease. Advances in knowledge in genetics and molecular biology have led to the development of novel treatments targeting specific molecules. Todate, the results obtained with these new drugs have not been superior to those observed with conventional chemotherapy. In the present article we review some of the new therapies for pancreatic cancer such as the use of inhibitors of epidermal growth factor receptors, the inhibitors of metalloproteases of the extracellular matrix, the inhibitors of farnesyltransferase, and the use of anti-angiogenic treatment. MLA is supported by grant 01/9563 from the Instituto de Salud Carlos III, Madrid, Spain.     

影响安罗替尼临床疗效和预后的多因素分析

目的:观察经过安罗替尼治疗的不同瘤种恶性肿瘤患者的相关指标,探索影响安罗替尼临床疗效和预后的因素。方法:2018年7月至2019年12月使用安罗替尼治疗的晚期恶性肿瘤患者101例,取患者治疗过程中的血液,用ELISA法和实时荧光定量PCR法检测血清中VEGF、FGF、c-Kit、Bcl-2、PARP蛋白含量和mRNA相对表达量。另一方面,评价安罗替尼的疗效和随访观察PFS。结果:全部病例的总的中位PFS为4.8月（95%CI:3.8~5.8）,PR 18例（17.82%）,SD 61例（60.40%）,PD 22例(21.78%),ORR 17.82%,DCR 78.22%。TSH升高患者的DCR较未发生者高（P=0.039 0）。既往曾使用铂类化疗者较未使用者具有更长的中位PFS（5.3月 vs 3.6月,Log-rank P=0.038 0）,TSH升高患者较未升高患者的中位PFS显著延长（6.4月 vs 4.0月,Log-rank P=0.046 0）,发生手足综合征者中位PFS较未发生者显著延长（11.4月 vs 4.2月,Log-rank P=0.021 0）。血清bFGF的蛋白含量（P=0.009 0）及Bcl-2 mRNA相对表达量（P=0.012 0）是安罗替尼治疗晚期恶性肿瘤预后的独立影响因子。结论:血清bFGF含量及Bcl-2 mRNA表达量是安罗替尼疗效的独立影响因素;既往曾使用铂类、发生手足综合征和促甲状腺素升高者具有更长的PFS。     

多肽mPEG-SC20k-HM-3联合奥沙利铂对人肝癌细胞SMMC-7721裸鼠移植瘤的抑制作用

mPEG-SC_20k-HM-3是具有整合素亲和性的一种新型高效血管生成抑制多肽,为探索其与传统化疗药物联用的抗肿瘤活性,建立人肝癌细胞SMMC-7721裸鼠移植瘤模型,选用奥沙利铂为化疗药物,根据临床前抗肿瘤药效学方法评价联合用药的抑瘤作用,并用金氏公式判断两种药物的联合作用。结果显示,与单独用药相比,联合用药组的抑瘤效果更好,且均具有显著差异(P<0.05)。其中,奥沙利铂(7.5 mg/kg)联合mPEG-SC_20k-HM-3(73.4 mg/kg)的肿瘤抑制率为84.6%,抑制作用比单用奥沙利铂(7.5 mg/kg)和单用mPEG-SC_20k-HM-3(73.4 mg/kg)均有显著提高。根据金氏公式计算,其Q值为1.164(>1.15),可判定该组的用药组合表现出协同作用。结果表明,mPEG-SC_20k-HM-3与奥沙利铂单独给药均能抑制肿瘤生长,两者联用对肝细胞癌具有协同作用。     

RNA干扰酪氨酸激酶受体2对人脐静脉内皮细胞增殖的影响

目的研究抑制酪氨酸激酶受体2（Tie2）对内皮细胞凋亡和增殖活性的影响。方法采用RNA干扰技术,将含有Tie2基因的特异性短发夹状RNA（shRNA）片段的质粒转染入人脐静脉内皮细胞（HUVECs）,采用实时定量逆转录聚合酶链反应和免疫印迹法检测细胞中Tie2 mRNA和蛋白表达的改变;噻唑蓝比色分析（MTT）法检测细胞的生
长情况;显微镜下观察细胞凋亡情况。以转染了pGenesil-hk质粒组为阴性对照,未转染质粒组为空白对照。结果质粒转染入HUVECs后,实验组细胞中Tie2 mRNA和蛋白表达水平较阴性对照组和空白对照组下调,尤以转染后48 h Tie2的mRNA和蛋白表达水平下调更为明显（P<0.05）。MTT检测结果显示：实验组细胞的增殖活性受到明显抑制（P<0.05）。转染48 h后,实验组的细胞凋亡率明显高于两个对照组（P<0.05）。结论RNA干扰技术沉默Tie2基因可致Tie2基因表达下调后诱导HUVECs凋亡,并抑制HUVECs的增殖。     

Cancer gene therapy: developments to 2000

Cancer, at the molecular level, continues to be more thoroughly understood. With this understanding comes the opportunity for innovative therapeutic intervention. Gene therapy remains an attractive concept to treat cancer. However, a number of gene therapy clinical trials have now been reported and it is clear that barriers remain before gene therapy gains widespread clinical application. This article outlines current directions and novel developments in the field of cancer gene therapy, which attempt to overcome these obstacles.