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.     

Ultrasound with Microbubble Contrast Agent and Urokinase for Thrombosis

This study was aimed at assessing the effects of urokinase (UK) in combination with ultrasound and microbubbles in in vitro and in vivo thrombolytic therapy for the treatment of deep vein thrombosis (DVT). Thrombi with formation times of 1, 3, 7, 14 and 21 d were used for thrombolysis. Forty-five adult mongrel dogs were used to evaluate thrombosis in vivo. Both in vitro and in vivo analyses revealed that UK?+?microbubbles had the best effect among the combinations. Thrombolysis <7 d was more effective at a thrombolysis rate of about 50%, but the thrombolytic effect of thrombi >7 d was poor at thrombolysis rates <30%. Ultrasound?+?UK significantly increased the thrombolysis rate of thrombi <7 d. These results suggest that the combination of ultrasound with microbubble contrast agents and UK may have a synergistic effect on thrombolysis.     

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.     

丹参川芎嗪注射液联合尿激酶治疗急性心肌梗死的疗效观察

目的探讨丹参川芎嗪注射液联合注射用尿激酶治疗急性心肌梗死的临床疗效。方法选择2016年6月—2017年7月在宣城市中心医院治疗的急性心肌梗死患者65例为研究对象,按照患者所用治疗方案差异将所有患者分为对照组(33例)和治疗组(32例)。对照组给予注射用尿激酶,先静脉推注,50万单位加入到5%葡萄糖溶液20 m L中,而后更改为静脉滴注,50万单位加入到5%葡萄糖液500 m L中,1次/d。治疗组在对照组的基础上静脉滴注丹参川芎嗪注射液,10 m L加入到生理盐水500 m L中,1次/d。两组患者均连续治疗14 d。观察两组的临床疗效,比较两组的心功能指标和血管再通情况。结果治疗后,对照组和治疗组的总有效率分别为75.76%、87.50%,两组比较差异具有统计学意义(P0.05)。治疗后,两组左室射血分数(LVEF)、每搏心输出量(SV)均明显升高,中心静脉压(CVP)明显降低,同组治疗前后比较差异具有统计学意义(P0.05);且治疗组这些观察指标的改善程度明显优于对照组,两组比较差异具有统计学意义(P0.05)。治疗后,对照组和治疗组的血管再通率分别为57.58%、84.38%,两组比较差异具有统计学意义(P0.05)。结论丹参川芎嗪注射液联合注射用尿激酶治疗急性心肌梗死具有较好的临床疗效,可改善心脏功能,提高血管再通率,具有一定的临床推广应用价值。     

胞二磷胆碱联合尿激酶治疗急性脑梗死的临床研究

目的探讨急性脑梗死应用胞二磷胆碱联合尿激酶治疗的临床效果。方法选取2016年1月—2018年6月襄阳市中心医院收治的102例急性脑梗死患者,运用随机数字表法随机分成对照组和治疗组,每组各51例。对照组静脉滴注注射用尿激酶,100万U注射用尿激酶溶于100 mL生理盐水,持续静脉滴注30 min,单次给药。治疗组在对照组治疗基础上静脉滴注胞二磷胆碱注射液,0.75 g加入150 mL生理盐水充分稀释,给药时间应大于40 min,1次/d。两组均治疗两周。观察两组的临床疗效,比较两组患者治疗前后血小板参数、国立卫生研究院卒中量表(NIHSS)评分、凝血–纤溶指标、脑血流动力学参数的变化情况。结果治疗后,对照组和治疗组的总有效率分别为76.5%、92.2%,两组比较差异具有统计学意义(P0.05)。治疗后,两组血小板计数(PLC)显著增加,但血小板最大聚集率(PAGT_(max))值和NIHSS评分均较治疗前显著减少,同组治疗前后比较差异有统计学意义(P0.05);治疗后,治疗组PLC值高于对照组,PAGT_(max)值和NIHSS评分低于治疗组,两组比较差异具有统计学意义(P0.05)。治疗后,两组纤维蛋白原(FIB)、D-二聚体(D-D)、外周阻力(R_v)水平均显著下降,两组平均血流速度(V_(mean))和平均血流量(Q_(mean))值较治疗前均显著增高,同组治疗前后比较差异有统计学意义(P0.05);治疗后,治疗组FIB、D-D、R_v水平显著低于对照组,V_(mean)和Q_(mean)显著高于对照组,两组比较差异具有统计学意义(P0.05)。结论胞二磷胆碱联合尿激酶治疗急性脑梗死具有较好的临床疗效,可有效抑制患者体内血小板活性,纠正凝血–纤溶系统紊乱,维持脑血流动力学稳定,减少神经功能缺损,具有一定的临床推广应用价值。     

立体定向靶向脑室内注射尿激酶治疗脑室出血铸型疗效分析

目的探讨立体定向靶向注射尿激酶治疗脑室出血铸型的疗效。方法采用立体定向靶向穿刺单侧或双侧脑室,后予以脑室内注射尿激酶行纤溶治疗,拔管后联合腰大池持续引流,预后采用ADL评定。结果全部未见颅内感染及再出血等并发症,ADLⅠ级2例,ADLⅡ级8例,ADLⅢ级8例,ADLⅣ级4例,ADLⅤ级2例,自动出院2例。结论立体定向靶向脑室内注射尿激酶治疗脑室出血铸型是挽救生命的一种重要方法,其方法安全、有效。     

rt-PA联合尿激酶静脉溶栓治疗急性脑梗死的疗效及安全性研究

目的观察重组组织型纤溶酶原激活剂联合尿激酶静脉溶栓治疗急性脑梗死的疗效与安全性。方法选择前循环急性脑梗死患者171例,联合溶栓组46例,单用rt-PA组39例,单用尿激酶组34例,对照组52例。观察治疗前及治疗后14d NIHSS评分,同时观察再梗死率、脑出血率及死亡率。结果 4组治疗后14d分别和治疗前比较,差异有统计学意义(P<0.05);3个溶栓组与对照组14d有效率差异有统计学意义(P<0.05);4组再发脑梗死率、死亡率差异无统计学意义(P>0.05),联合溶栓组脑出血率与单用rt-PA组及单用尿激酶组差异均有统计学意义(P<0.05)。结论 rt-PA联合尿激酶治疗急性脑梗死是安全有效的。     

中分子羟乙基淀粉联合小剂量尿激酶治疗急性脑分水岭梗死的疗效观察

目的 观察中分子羟乙基淀粉联合小剂量尿激酶治疗急性脑分水岭梗死（CWI）患者的临床疗效.方法 68例CWI患者随机分为两组,对照组（n=34）在常规治疗措施基础上加用小剂量尿激酶,观察组（n=34）在常规治疗措施基础上加用中分子羟乙基淀粉、小剂量尿激酶联合治疗.比较两组治疗总有效率,治疗前后神经功能缺损程度（NIHSS）和Barthel指数变化情况.结果 观察组治疗总有效率（88.24％）明显高于对照组（67.65％） （x2=7.383,P＜0.05）;两组患者治疗前NIHSS、Bar-thel比较差异无统计学意义（P＞0.05）,随治疗时间延长,NIHSS逐渐降低,Barthel逐渐升高,且观察组降低和升高较对照组更为显著（P＜0.05）;两组治疗期间未见严重不良反应发生.结论 中分子羟乙基淀粉联合小剂量尿激酶治疗CWI疗效显著,可明显改善神经功能缺损程度和日常生活能力,值得临床推广应用.     

尿激酶早期溶栓治疗急性脑梗死

目的 观察早期应用尿激酶（UK）治疗急性脑梗死的临床效果。方法 选择53例发病在6h内的急性脑梗死，随机分成UK组（28例）和对照组（25例），UK组UK150万U30min内静脉滴注，对照组不用UK，其余两组治疗相同。评价治疗前和治疗后14d神经功能缺损程度评分。结果 UK组总有效率为92．8％，对照组84％（P〈0．05）；UK组基本痊愈率为60.7％，与对照组基本痊愈率24％相比差异有统计学意义（P〈0．01）。个别病例出现轻度皮肤、粘膜出血，停药后症状消失，未发现严重不良反应。结论 UK治疗急性脑梗死临床疗效确切、安全，不良反应轻微。     

Independent validation of stromal uPA in ABCSG-08: Level 1b evidence for the prognostic value of uPA immunohistochemistry

PurposeTo validate the prognostic role of urokinase-type plasminogen-activator (uPA) and plasminogen activator inhibitor type-1 (PAI-1) protein expression in FFPE archived tumor samples when assessed by immunohistochemistry.Patients and methodsFresh-frozen, paraffin-embedded (FFPE) samples from 303 postmenopausal women with hormone receptor-positive, early breast cancer were investigated. The patients had received 5 years of endocrine therapy in the prospectively randomized ABCSG-8 trial. Immunohistochemistry for stromal uPA and PAI-1 protein expression was correlated with distant recurrence-free survival (DRFS) and overall survival (OS).ResultsWe detected stromal uPA in 132 of 297 tumors (44.4%) and stromal PAI-1 expression in 74 out of 299 samples (24.7%). Co-expression of uPA and PAI-1 was present in 48 of 294 (16.3%) cases. Neither uPA nor PAI-1 expression was associated with tumor size, age, nodal status, grading, or quantitative receptor status. Patients whose tumor stroma expressed uPA protein had a significantly shorter DRFS (adjusted HR for relapse: 2.78; 95% CI 1.31–5.93; p = 0.008 Cox regression analysis) than women without uPA expression. No such association was seen for PAI-1 and the uPA/PAI1 ratio. After a median follow-up of 5.6 years, women with uPA-positive tumors demonstrated significantly shorter DRFS (93.3% vs. 84.8%; p < 0.013 log-rank test), and tended to have a worse OS (83.0% vs. 77.3%; p = 0.106) compared to women with uPA negative tumors.ConclusionThis independent validation in archived tumor samples from a large prospective randomized trial confirms the clinical utility of stromal uPA evaluation by immunohistochemistry. This provides level 1b evidence for the prognostic role of stromal uPA in women with endocrine-responsive early breast cancer.