uPAR Controls Vasculogenic Mimicry Ability Expressed by Drug-Resistant Melanoma Cells

Malignant melanoma is a highly aggressive skin cancer characterized by an elevated grade of tumor cell plasticity. Such plasticity allows adaptation of melanoma cells to different hostile conditions and guarantees tumor survival and disease progression, including aggressive features such as drug resistance. Indeed, almost 50% of melanoma rapidly develop resistance to the BRAFV600E inhibitor vemurafenib, with fast tumor dissemination, a devastating consequence for patients’ outcomes. Vasculogenic mimicry (VM), the ability of cancer cells to organize themselves in perfused vascular-like channels, might sustain tumor spread by providing vemurafenibresistant cancer cells with supplementary ways to enter into circulation and disseminate. Thus, this research aims to determine if vemurafenib resistance goes with the acquisition of VM ability by aggressive melanoma cells, and identify a driving molecule for both vemurafenib resistance and VM. We used two independent experimental models of drug-resistant melanoma cells, the first one represented by a chronic adaptation of melanoma cells to extracellular acidosis, known to drive a particularly aggressive and vemurafenib-resistant phenotype, the second one generated with chronic vemurafenib exposure. By performing in vitro tube formation assay and evaluating the expression levels of the VM markers EphA2 and VE-cadherin by Western blotting and flow cytometer analyses, we demonstrated that vemurafenib-resistant cells obtained by both models are characterized by an increased ability to perform VM. Moreover, by exploiting the CRISPR-Cas9 technique and using the urokinase plasminogen activator receptor (uPAR) inhibitor M25, we identified uPAR as a driver of VM expressed by vemurafenib-resistant melanoma cells. Thus, uPAR targeting may be successfully leveraged as a new complementary therapy to inhibit VM in drug-resistant melanoma patients, to counteract the rapid progression and dissemination of the disease.     

Non-viral transfer of BDNF and uPA stimulates peripheral nerve regeneration

Peripheral nerves connect brain and spinal cord with the extremities and inner organs, and nerves injury can lead the disability and social exclusion. Growth factors and other natural stimulators of regeneration processes look very promising as future medicines. In our study, we tested the influence of genetic constructions that contain genes of brain-derived neurotrophic factor and urokinase plasminogen activator on nerve's structure and function after traumatic and ischemic injuries. Injection of pVax1-hBDNF and pVax1-muPA after traumatic injury led to better restoration of nerve's structure and function compared to similar parameters of control group mice. In ischemic injury model pVax1-hBDNF and pVax1-muPA slowed and reduced the damage progression and stimulated nerve regeneration as well. However, the treatment with pVax1-muPA was less effective after the traumatic injury. As we chose a non-viral method of gene delivery during our study the optimal conditions of plasmid intramuscular delivery were also determined.     

深静脉途径尿激酶溶栓治疗下肢深静脉血栓形成的疗效观察

目的 观察不同给药途径治疗下肢深静脉血栓形成的效果.方法 40例下肢深静脉血栓形成患者分为两组,A组20例,给予尿激酶50万IU,通过患肢深静脉途径给药;B组20例,给予尿激酶50万IU,通过上肢静脉途径给药.结果 A组有效率为95%,B组有效率为55%,两组比较P〈0.05,有统计学意义.结论 经患肢深静脉途径给药治疗下肢深静脉血栓形成可取得良好疗效,方法简单、安全.     

阿替普酶与尿激酶治疗急性脑梗死疗效的比较研究

目的对比阿替普酶和尿激酶治疗急性脑梗死的疗效。方法收集哈尔滨医科大学附属第一医院神经内科静脉溶栓治疗的急性脑梗死患者158例,根据所用溶栓药物分为两组:rt-Pa组患者53例。UK组患者105例,其中发病4.5 h内患者80例,构成4.5 UK组。神经功能缺失评定采用NIHSS评分。分别观察溶栓后2 h、24h、7 d、10 d疗效。结果(1)rt-Pa组在2 h、24 h疗效优于UK组(P0.05),rt-Pa组和UK组在7 d和10 d疗效无明显差异(P0.05)。(2)rt-Pa组在2 h、24 h疗效优于4.5 UK组(P0.05),rt-Pa组和4.5UK组在7 d和10 d疗效无明显差异(P0.05)。结论阿替普酶在溶栓后24 h内疗效优于尿激酶,但二者的短期预后相当。     

尿激酶溶栓治疗脑血栓40例疗效评估报道

目的：对脑血栓治疗中尿激酶溶栓疗法起到的临床作用进行分析探讨。方法选取2012年7月～2014年12月同期于我院进行脑血栓治疗的患者40例,将其分为实验组和对照组。对对照组患者给予低分子肝素进行治疗,对实验组患者给予尿激酶进行溶栓治疗。治疗结束之后对两组患者的治理效果及缺损评分进行比较。结果实验组的总体有效率为80.00%较对照组的60.00%具有优势,实验组缺损评分低于对照组,数据具有统计学差异（P＜0.05）。结论对脑血栓患者采取尿激酶进行溶栓治疗能有效治愈患者疾病。     

持续股动脉输注脲激酶治疗糖尿病足的临床研究

目的 观察采用微量动脉泵(微量泵)持续股动脉输注脲激酶治疗糖尿病足溃疡的临床疗效.方法 选择2型糖尿病并发足溃疡患者(Wagner 3级及以下者),无内脏出血倾向,无严重心肾肝功不全,在抗感染、降糖治疗及局部创面处理的同时,给予微量泵持续股动脉输注脲激酶治疗.结果 28例糖尿病足溃疡全部愈合,平均住院9±5 天.结论 微量泵持续股动脉输注脲激酶是治疗糖尿病足溃疡的一种有效方法,可减少截肢,缩短住院时间.必须掌握本疗法的适应证和禁忌证.     

乙酰肝素酶及其相关因子mRNA在白血病细胞中的表达

目的 观察乙酰肝素酶(HPA)及其相关因子mRNA在白血病细胞中的表达,研究白血病细胞表面止血平衡改变.方法 选取白血病患者40例(研究组),缺铁性贫血患者20例(对照组).采用半定量反转录聚合酶链反应(RT-PCR)方法测定受试者骨髓单个核细胞组织因子(TF)、尿激酶型纤溶酶原激活物受体(uPAR)、HPA mRNA的含量.结果 研究组骨髓中单个核细胞TF、uPAR、HPAmRNA的表达与对照组相比均升高(均P＜0.05),35例初发白血病患者中急性髓系白血病(AML)组(23例)骨髓中单个核细胞TF、uPAR、HPA mRNA高于慢性粒细胞白血病(CML)组(4例)、慢性淋巴细胞白血病(CLL)组(3例)、急性淋巴细胞白血病(ALL)组(5例)(对照组、AML组、CML组、ALL组、CLL组TF分别为0.66±0.03、2.56±0.37、1.33±0.06、1.93±0.08、1.23±0.07,uPAR分别为0.53士0.02、1.89±0.26、0.92±0.15、1.44±0.01、1.19±0.16,HPA分别为2.37±0.24、1.42±0.13、1.68±0.09、1.54±0.12,均P＜0.05).结论 白血病细胞中HPA、TF、uPAR mRNA表达增高;不同类型白血病表达水平不同.提示TF、uPAR、HPA可能影响白血病细胞表面止血平衡,不同类型白血病细胞表面止血平衡改变不同.     

A hybrid protein of the amino-terminal fragment of urokinase and mutant plasminogen activator inhibitor-2 efficiently inhibits tumor cell invasion and metastasis

The urokinase plasminogen activator(uPA) system plays important roles in tumor cell invasion and metastasis. In the present study, we evaluated the effects of ATF-PAI2CD, a hybrid protein of the amino-terminal fragment of urokinase and mutant plasminogen activator inhibitor-2, on 95D cells in vitro and in vivo. Furthermore, our results support a current hypothesis that fusion protein blocks tumor invasion and motility by inhibiting localized pericellular proteolysis. Treatment of 95D cells with ATF-PAI2CD resulted in a dose-dependent decrease in tumor-cell invasion through matrigel, and ATF-PAI2CD was much more effective than PAI-2CD. In addition, extracellular regular protein kinase (ERK1/2) expression was downregulated and the adhesion ability to fibronectin was increased in 95D cells treated with the fusion protein, which was confirmed by cell adhesion assay. A high-concentration of ATF-PAI2CD caused a significant reduction in tumor volume and weight in BALB/c (nu/nu) mice female inoculated with human 95D cells (5×10⁶); the antitumor effects were significant, which demonstrated a 67.9±4.2% reduction in tumor growth compared with control mice. The number of lymphatic metastasis was significantly reduced in mice treated with high- and middle- concentrations of ATF-PAI2CD, whereas a low-concentration of ATF-PAI2CD failed to exhibit any antimetastatic effects. In conclusion, the results suggested that the hybrid protein has therapeutic potential for lung carcinoma and other tumors to inhibit tumor invasion and metastasis.     

尿激酶与阿替普酶对急性肺栓塞溶栓的疗效对比分析

目的：探讨尿激酶与阿替普酶在治疗急性肺栓塞溶栓的疗效。方法随机选取2014年1月-2015年1月该院收治的急性肺栓塞高危患者120例,将其随机分为对照组与观察组,每组60例,观察组应用阿替普酶治疗,对照组应用尿激酶治疗,两组均采用2 h静脉溶栓治疗,记录并分析两组患者溶栓前后的临床症状变化情况、CT肺动脉造影(CTPA)变化情况、不良反应的发生情况及患者死亡率。结果对照组溶栓的有效率为86.7%,出血率为11.7%,死亡率为1.7%;观察组溶栓的有效率为96.7%,出血率为13.3%,死亡率为1.7%,观察组溶栓的总有效率明显高于对照组,差异具有统计学意义(P<0.05)。观察组患者的出血率与死亡率和对照组比较,差异无统计学意义(P跃0.05)。结论阿替普酶溶栓治疗急性肺栓塞有效率高于尿激酶,但两种溶栓方案的死亡率与出血率无差异,因此在临床治疗中阿替普酶溶栓方案优于尿激酶。     

胸腔置管引流联合胸腔内注入不同剂量的尿激酶治疗复杂性肺炎旁胸腔积液的疗效探讨

目的：探讨不同剂量的尿激酶在治疗复杂性肺炎旁胸腔积液中的疗效。方法将48复杂性例肺炎旁胸腔积液患者随机分为大剂量尿激酶治疗组和小剂量尿激酶对照组,大剂量治疗组26例,小剂量对照组22例。大剂量治疗组首次胸腔抽液后注入含25万u尿激酶的生理盐水20 mL,小剂量对照组给予含尿激酶10万u的生理盐水20 mL。比较两组胸腔积液引流量、住院时间及治疗效果。结果在规则抗感染的治疗下,大剂量治疗组平均胸液引流量(1840±100)mL、平均住院时间为(20.4±2)d,大剂量治疗组总有效率92.3%;小剂量对照组平均胸液引流量(1420±90) mL,平均住院时间为(27.2±2)d,总有效率63.6%,比较两组平均胸液引流量、平均住院时间及总有效率差异有统计学意义(P<0.05)。不良反应两组无差异。结论胸腔引流后腔内应用大剂量尿激酶25万u,能显著增加胸水引流量,减少住院时间,疗效满意,且未出现明显的毒副作用。