MicroRNA-130a associates with ribosomal protein L11 to suppress c-Myc expression in response to UV irradiation

The oncoprotein c-Myc is essential for cell growth and proliferation while its deregulated overexpression is associated with most human cancers. Thus tightly regulated levels and activity of c-Myc are critical for maintaining normal cell homeostasis. c-Myc is down-regulated in response to several types of stress, including UV-induced DNA damage. Yet, mechanism underlying UV-induced c-Myc reduction is not completely understood. Here we report that L11 promotes miR-130a targeting of c-myc mRNA to repress c-Myc expression in response to UV irradiation. miR-130a targets the 3′-untranslated region (UTR) of c-myc mRNA. Overexpression of miR-130a promotes the Ago2 binding to c-myc mRNA, significantly reduces the levels of both c-Myc protein and mRNA and inhibits cell proliferation. UV treatment markedly promotes the binding of L11 to miR-130a, c-myc mRNA as well as Ago2 in cells. Inhibiting miR-130a significantly suppresses UV-mediated c-Myc reduction. We further show that L11 is relocalized from the nucleolus to the cytoplasm where it associates with c-myc mRNA upon UV treatment. Together, these results reveal a novel mechanism underlying c-Myc down-regulation in response to UV-mediated DNA damage, wherein L11 promotes miR-130a-loaded miRISC to target c-myc mRNA.     

Combination of circulating tumor cell enumeration and tumor marker detection in predicting prognosis and treatment effect in metastatic castration-resistant prostate cancer

Although circulating tumor cell (CTC) enumeration in peripheral blood has already been validated as a reliable biomarker in predicting prognosis in metastatic castration-resistant prostate cancer (mCRPC), patients with favorable CTC counts (CTC < 5/7.5 ml) still experience various survival times. Assays that can reduce patients'' risks are urgently needed. In this study, we set up a real-time quantitative polymerase chain reaction (RT-qPCR) method to detect epithelial-mesenchymal transition (EMT) and stem cell gene expression status in peripheral blood to validate whether they could complement CTC enumeration. From January 2013 to June 2014 we collected peripheral blood from 70 mCRPC patients and enumerated CTC in these blood samples using CellSearch system. At the same time, stem cell-related genes (ABCG2, PROM1 and PSCA) and EMT-related genes (TWIST1 and vimentin) were detected in these peripheral blood samples using an RT-qPCR assay. Patient overall survival (OS) and treatment methods were recorded in the follow-up. For patients who received first-line chemotherapy, docetaxel plus prednisone, PSA progression-free survival (PSA-PFS) and PSA response rate were recorded. At the time of analysis, 35 patients had died of prostate cancer with a median follow-up of 16.0 months. Unfavorable CTC enumerations (CTC ≥5/7.5 ml) were predictive of shorter OS (p = 0.01). Also, positive stem cell gene expression indicated poor prognosis in mCRPC patients (p = 0.01). However, EMT gene expression status failed to show any prognostic value in OS (p = 0.78). A multivariate analysis indicated that serum albumin (p = 0.04), ECOG performance status (p < 0.01), CTC enumeration (p = 0.02) and stem cell gene expression status (p = 0.01) were independent prognostic factors for OS. For the 40 patients categorized into the favorable CTC enumeration group, positive stem cell gene expression also suggested poor prognosis (p < 0.01). A combined prognostic model consisting of stem cell gene expression and CTC enumeration increased the concordance probability estimated value from 0.716 to 0.889 in comparison with CTC enumeration alone. For patients who received docetaxel plus prednisone as first-line chemotherapy, positive stem cell gene expression suggested a poor PSA-PFS (p = 0.01) and a low PSA response rate (p = 0.008). However, CTC enumeration and EMT gene expression status did not affect PSA-PFS or PSA response rates. As a result, detection of peripheral blood stem cell gene expression could complement CTC enumeration in predicting OS and docetaxel-based treatment effects in mCRPC patients.     

大鼠脑内囊出血模型的构建及其评价

背景:稳定准确的动物模型是研究出血性脑血管病的必要工具和基础。目的:建立和评价大鼠脑内囊出血模型。设计:随机对照动物实验。单位:徐州医学院第二附属医院;南京医科大学第一附属医院。材料:实验于2002-05/11在南京医科大学动物实验中心完成。35只SD大鼠随机分为两组:实验组30只,假手术组5只。方法:①通过立体定向术向实验组大鼠脑内囊注入自体血制成脑内囊出血模型。②按ZeaLonga5分制神经病学评分标准评分,观察大鼠躯体感觉及运动功能。③大鼠在麻醉状态下于术前及术后检测体感诱发电位。④测定体感诱发电位后,将大鼠麻醉后处死,取脑制备切片,苏木精-伊红染色,以最大病灶处光镜观察血肿及组织形态学的改变。主要观察指标:①两组大鼠神经功能评分。②两组大鼠体感诱发电位各波潜伏期。③两组大鼠脑组织形态学观察。结果:35只大鼠均进入结果分析。①以出现明显偏瘫为造模成功,本实验成功率为93.3%(28/30),实验组神经病学评分为(2.74±0.46)分,与假手术组(0分)比较,差异有显著性(P<0.05)。②体感诱发电位显示,实验组术后各波潜伏期较术前和假手术组明显延迟[P1:(15.72±0.78)ms,(10.69±0.52)ms,(10.73±0.48)ms;N1:(17.95±1.27)ms,(13.21±1.31)ms,(13.34±1.27)ms;N2:(21.16±1.62)ms,(15.42±1.46)ms,(15.58±1.44)ms;N3:(24.86±1.58)ms,(18.72±1.76)ms,(18.99±1.67)ms,P<0.05]。③假手术组仅见针道周围散在红细胞,无出血灶;实验组病理形态学表现为左侧内囊区不规则或椭圆形血凝块,大致有一个低倍视野范围,出血灶边缘脑组织疏松水肿,病变明显重于假手术组。结论:用立体定向术回注自体血制成大鼠脑内囊出血模型更接近临床脑出血,且方法简便,重复性好。     

聚乳酸-O-羧甲基壳聚糖纳米粒子对异种移植猪肝细胞凋亡的抑制作用

目的：观察聚乳酸-O-羧甲基壳聚糖纳米粒子对猪肝细胞异种移植后移植肝细胞凋亡的抑制作用. 方法：实验于2004-03/2005-03在南通大学神经再生实验室完成,选用中国实验用小型猪（n=5）及SD大鼠（n=123）.①实验分组及方法：采用原位胶原酶循环灌注法分离猪肝细胞.D-氨基半乳糖腹腔内注射制作大鼠急性肝衰竭模型,按随机数字表法分成3组（n=41）,分别在单纯肝细胞移植组、胶原肝细胞移植组、纳米胶原肝细胞移植组急性肝衰竭大鼠腹腔内移植震荡培养24 h的猪肝细胞悬液、Ⅰ型胶原凝胶固定培养24 h的猪肝细胞、Ⅰ型胶原凝胶包埋的聚乳酸-O-羧甲基壳聚糖纳米粒子培养24 h的猪肝细胞.②实验评估：观察移植肝细胞的病理变化、培养和移植肝细胞的凋亡率、坏死率及活率. 结果：①各组移植肝细胞的活率及凋亡率：纳米胶原肝细胞移植组移植肝细胞的存活时间最长.肝细胞体外培养1 d后,3组肝细胞均存在不同程度的凋亡,以单纯肝细胞移植组凋亡率最高,纳米胶原肝细胞移植组最低.胶原肝细胞移植组、纳米胶原肝细胞移植组的移植肝细胞凋亡率随着移植时间的延长呈缓慢上升趋势,但较单纯肝细胞移植组为低;移植后1,2 d,胶原肝细胞移植组、纳米胶原肝细胞移植组的移植肝细胞活率明显高于单纯肝细胞移植组（P＜0.05）;移植后3,5 d时,纳米胶原肝细胞移植组移植肝细胞凋亡率明显低于胶原肝细胞移植组（P＜0.05）,活率则明显高于胶原肝细胞移植组（P＜0.05）.②各组移植肝细胞的坏死率：移植后1 d,胶原肝细胞移植组移植肝细胞坏死率明显上升;移植后2 d,3组肝细胞坏死率均有不同程度的上升,胶原肝细胞移植组、纳米胶原肝细胞移植组明显高于单纯肝细胞移植组（P＜0.05）,胶原肝细胞移植组高于纳米胶原肝细胞移植组（P＜0.05）.移植后3 d,单纯肝细胞移植组肝细胞坏死率大幅度上升,胶原肝细胞移植组、纳米胶原肝细胞移植组无明显变化,但显著低于单纯肝细胞移植组（P＜0.05）,胶原肝细胞移植组、纳米胶原肝细胞移植组之间差异无显著性意义（P＞0.05）.移植后5 d,胶原肝细胞移植组、纳米胶原肝细胞移植组肝细胞坏死率进一步上升,纳米胶原肝细胞移植组显著低于胶原肝细胞移植组（P＜0.05）. 结论：聚乳酸-O-羧甲基壳聚糖纳米粒子有抑制培养和移植肝细胞凋亡、提高移植肝细胞活率的作用,聚乳酸-O-羧甲基壳聚糖纳米粒子和Ⅰ型胶原结合可增强抗培养和移植肝细胞凋亡的能力,延长移植肝细胞的生存时间.     

对流行性出血热病人入量指导及饮食干预的效果观察

流行性出血热又称肾综合征出血热,是由汉坦病毒引起的以鼠类为传染源的自然疫源性疾病。典型病例有发热、出血和肾脏损害三大主征和发热期、低血压休克期、少尿期、多尿期、恢复期“五期”特点,病人各期对入量和饮食的要求均不相同。我院对120例住院病人进行了入量指导及饮食干     

自体骨髓基质源干细胞在损伤脑区及其他脑区的分布

背景:近来骨髓基质细胞多向分化潜能的发现,特别是骨髓基质细胞向神经干细胞方向分化的成功,为干细胞移植治疗神经系统损伤提供了可能。目的:观察自体骨髓基质源干细胞对脑损伤的治疗作用。设计:以BrdU标记的自体骨髓基质源干细胞在模型动物损伤脑区和其他脑区的分布为观察对象的前瞻性研究。单位:北京军区总医院。材料:实验于1999-10/2002-02在北京军区总医院完成。取1.5～3.0岁成年家犬22只,雌雄不限,体质量(13±2)kg。随机分为假手术组6只,常规治疗组8只,骨髓基质源神经干细胞组8只。方法:对犬骨髓基质细胞进行采集、分离、体外扩增、标记和移植用细胞培养。假手术组仅做皮肤切口及颅骨锥孔,不致中脑损伤,于麻醉清醒后行正常进食。常规治疗组行中脑被盖区毁损后行三磷酸腺苷20mg,辅酶A50u,加入生理盐水中静脉滴注,2次/d,共3d。骨髓基质源神经干细胞组行中脑被盖区毁损,然后给予骨髓基质细胞源神经干细胞1mL和脑脊液1mL的混合液,注入脑室,1次/d,共3d。术后第1,7,14,28天4个时间点观察骨髓基质源干细胞在损伤脑区及其他脑区的分布情况。主要观察指标:观察骨髓基质源干细胞在损伤脑区及其它脑区的分布情况。结果:22只动物均进入结果分析。①在术后第28天:骨髓基质源神经干细胞组BrdU阳性细胞明显高于假手术组和常规治疗组[(15.4±2.4,0.5±0.5,0.5±0.5)个]。②骨髓基质源神经干细胞组术后第14天脑片BrdU阳性细胞数:中脑的伤侧明显高于对侧[(15.5±3.3,5.0±1.5)个];额叶的伤侧高于对侧[(1.6±0.6,1.5±0.7)个]。结论:由于骨髓基质源神经干细胞移植组BrdU阳性细胞数在脑损伤区的分布明显多于对侧非伤区和两个对照组的同部位。提示骨髓基质源干细胞可向损伤脑区迁移,能对脑干损伤的恢复有一定促进作用。     

蛛网膜下腔出血后不同血液成分与脑血管痉挛及免疫炎症反应的关系

目的：探讨不同血液成分与蛛网膜下腔出血(SAH)后脑血管痉挛(CVS)及血管壁免疫炎症反应之间的关系。方法：分别将全血、孵育全血、富血小板血浆、孵育血浆等分别注入动物蛛网膜下腔，再于不同的时间段处死实验动物观察基底动脉的痉挛情况并测定血管壁中血管细胞粘附分子1(VCAM—1)的表达。结果：富血小板血浆、孵育血浆未引起基底动脉明显痉挛和VCAM—1的上调，全血在第2次注入后第3天出现基底动脉痉挛和VCAM—1表达的上调，孵育全血第1次注入后6小时就引起明显的基底动脉痉挛和VCAM—1的表达上调。结论：血管痉挛程度与血管壁VCAM—1的表达程度相一致；陈旧的红细胞或其裂解产物是引起SAH后血管痉挛和炎症反应主要成分。     

医用一次性瓶袋可转换型输液器的研制及应用

目前,临床输注液体的包装更新换代,输注液体已换成袋包装,故临床中袋式与瓶式液体(或药物)交替输注存在以下题:①输注袋式液体时输液器不需排气管;②输瓶式液体时输器需排气管。为此,我们研制出一种医用一次性瓶袋可转换输液器,解决了目前临床瓶式输液与袋式输液互相转换出现问