人重组可溶性肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导急性髓系白血病细胞凋亡的实验研究

为了研究肿瘤坏死因子相关凋亡诱导配体(TRAIL)对急性髓系白血病(AML)细胞的作用，通过MTT比色法测定不同浓度TRAIL对39例AML患者(病例组)和21例健康人(对照组)的骨髓或外周血单个核细胞的杀伤率。用流式细胞术检测细胞凋亡。结果发现，不同浓度TRAIL对AML细胞均有不同程度的杀伤作用，而对正常细胞无此作用。结论：TRAIL能通过诱导细胞凋亡而杀伤AML细胞，是一种有望应用于临床白血病治疗的新型生物制剂。     

Restricted Expression of an Adenoviral Vector Encoding Fas Ligand (CD95L) Enhances Safety for Cancer Gene Therapy

Gene transfer of Fas ligand (CD95L) using adenoviral vectors has been shown to generate apoptotic responses and potent inflammatory reactions that can be used to induce the regression of malignancies in vivo, but these vectors also cause significant hepatotoxicity that may limit their clinical utility. Here we describe an adenoviral vector encoding CD95L with restricted gene expression that reduces its toxicity in vivo. Preclinical efficacy and gene expression studies of lineage-restricted CD95L adenoviral vectors were performed. To enhance its cytotoxicity and reduce potential systemic effects, a noncleavable CD95L was made by deleting a segment containing the cleavage site (CD95LδQP). Higher CD95L expression of this mutant was observed on the tumor cell surface, together with a reduction in the release of soluble CD95L. This CD95L cleavage mutant was then expressed under control of a smooth muscle-specific promoter, SM22α, and analyzed for its ability to suppress the growth of tumors of smooth muscle origin in vivo. Growth of human leiomyosarcomas but not gliomas was inhibited after ADV gene transfer into tumor-bearing immunodeficient mice. In contrast to viral promoters, in which mortality was uniformly seen after injection of 10¹² particles, no significant hepatic injury or systemic toxicity was observed in mice, and the maximum tolerated dose was increased ≥10- to 100-fold. These findings suggest that restricted specificity of adenoviral CD95L gene expression enhances the safety of this approach for cancer gene therapy.     

Differential Induction of Apoptosis by Fas–Fas Ligand Interactions in Human Monocytes and Macrophages

Human monocytes undergo spontaneous apoptosis upon culture in vitro; removal of serum from the media dramatically increases the rate of this process. Monocyte apoptosis can be significantly abrogated by the addition of growth factors or proinflammatory mediators. We have evaluated the role of the endogenous Fas–Fas ligand (FasL) interaction in the induction of this spontaneous apoptosis and found that a Fas–immunoglobulin (Ig) fusion protein, an antagonistic anti-Fas monoclonal antibody and a rabbit anti-FasL antibody all greatly reduced the onset of apoptosis. The results indicate that spontaneous death of monocytes is mediated via an autocrine or paracrine pathway. Treatment of the cells with growth factors or cytokines that prevented spontaneous apoptosis had no major effects on the expression of Fas or FasL. Additionally, monocyte-derived macrophages were found to express both Fas and FasL but did not undergo spontaneous apoptosis and were not sensitive to stimulation by an agonistic anti-Fas IgM. These results indicate that protective mechanisms in these cells exist at a site downstream of the receptor–ligand interaction.     

辛伐他汀和普罗布考对TNF-α诱导的内皮细胞凋亡的影响

【目的】探讨辛伐他汀和普罗布考对TNF-α诱导人脐静脉内皮细胞（HUVEC）凋亡的影响及其影响的量效关系和时效关系。【方法】体外培养HUVES,取对数生长期的细胞分为四个组：①TNF-α干预的浓度依赖性组（Ac组）[按浓度分为Acl（Ong／mL）,Ac2（1ng／mL）,Ac3（10ng／mL）,Ac4（25ng／mL）];②TNF-α干预的时间依赖性组（At组）[按时间分为Atl（Oh）,At2（12h）,At3（24h）,At4（48h）];③10ng／mLTNF_a＋不同浓度辛伐他汀干预组（B组）[浓度分为B1（O μmol／L）,B2（O．01 μmol／L。）,B3（O．1 μmol／L。）,B4（1．0t μmol／L）];④1O ng／mL TNF-a不同浓度普罗布考干预组（c组）[浓度分为C1 （0 μmol／L。）,C2（20 μmol／L）,c3（40 μmol／L）,C4（80 μmol／L）]。以上四组均干预12h后,用流式细胞术检测各组凋亡率。【结果】①Ac组凋亡率分别为：Acl（O．81±0．25％）,Ac2（2．35±0．04％）,Ac3（4．54±0．32％）,Ac4（6．17±0．28％）,各组之间存在统计学差异（P〈0．01）。②At组凋亡率分别为,Atl（0．87±o．35％）,At2（4．53±0．32％）,At3（7．45±1．97％）,At4（10．92±1．27％）,各组间存在统计学差异（P〈O．01）。③B组凋亡率分别为,B1（4．46±0．53％）,B2（1．38±0．12％）,B3（2．89±0．27％）,B4（3．65±0．08％）,各干预组与对照组之间存在统计学差异（P〈O．01）。④C组细胞凋亡率分别为,C1（4．60±0．64％）,C2（2．61±0．18％）,C3（2．21±0．22％）,c4（1．28±0．34％）,各干预组与对照组之间存在统计学差异（P〈0．01）。【结论】①TNF-α可以诱导内皮细胞凋亡,且存在浓度依赖性和时．：良赖性。②辛伐他汀可以抑制TNF-α口诱导的内皮细胞凋亡,但无浓度依赖性。③普罗布考可以抑制TNF-α诱导的内皮细胞凋亡,但无浓度依赖性。     

AAV8-mediated Gene Therapy Prevents Induced Biochemical Attacks of Acute Intermittent Porphyria and Improves Neuromotor Function

Acute intermittent porphyria (AIP), an autosomal dominant hepatic porphyria due to half-normal hydroxymethylbilane synthase (HMB-synthase) activity, is manifested by life-threatening acute neurological attacks that are precipitated by factors that induce heme biosynthesis. The acute attacks are currently treated with intravenous hemin, but a more continuous therapy is needed, particularly for patients experiencing frequent attacks. Thus, a recombinant AAV8-based serotype vector expressing murine HMB-synthase driven by liver-specific regulatory elements was generated and its effectiveness to prevent the biochemical induction of an acute attack was evaluated in an AIP mouse model. Intraperitoneal administration of the adeno-associated viral (AAV) vector resulted in a rapid and dose-dependent increase of HMB-synthase activity that was restricted to the liver. Stable expression of hepatic HMB-synthase was achieved and wild-type or greater levels were sustained for 36 weeks. When heme synthesis was periodically induced by a series of phenobarbital injections, the treated mice did not accumulate urinary δ-aminolevulinic acid (ALA) or porphobilinogen (PBG), indicating that the expressed enzyme was functional in vivo and prevented induction of the acute attack. Further, rotarod performance and footprint analyses improved significantly. Thus, liver-directed gene therapy provided successful long-term correction of the hepatic metabolic abnormalities and improved neuromotor function in the murine model of human AIP.     

FAS,FASL及Bcl-2在化疗药物诱导RMA细胞凋亡过程中的表达

本研究通过测定RMA细胞Fas,FasL和Bcl-2表达的变化，探讨其在细胞凋亡过程中的作用。在培养的小鼠T淋巴瘤RMA细胞系中加化疗药物地塞米松（DEX）、足叶乙甙（VP－16）、三氧化二砷（As2O3)及维甲酸（ATRA）以及培养细胞中先分别与细胞国子IL－2，IL－6或GM－CSF共同培养后再加入上述药物，观察对细胞凋亡的影响及细胞凋亡过程中Fas,FasL mRNA,Fas及Bcl-2抗原的表达。DEX和VP－16能上调Fas和FasL表达，促进细胞凋亡，Bcl-2表达无变化。ATRA可下调Bcl-2表达，但不影响Fas和FasL系统，也未观察到有细胞凋亡。As2O3可以诱导细胞凋亡,但Fas,FasL及Bcl-2表达均无变化。提示不同药物对一种细胞可能通过不同的信号途径诱导调亡，而Fas系统诱导的细胞凋亡需要Fas和FasL共同参与。单独用IL－2，IL－6或GM－CSF虽然使Fas蛋白增加，但不引起细胞凋亡；如同时并用IL－2和IL－6则Fas和FasL表达均上升，并诱导细胞凋亡。上述细胞因子与化疗药物并用时可减低药物量，促进药物的凋亡诱导作用。在无FasL表达情况下，抗Fas单克隆抗体能诱导RMA细胞凋亡。实验结果表明，细胞因子与化疗药物可协同作用诱导细胞凋亡，Fas-FasL系统参与DEX和VP－16诱导的RMA细胞凋亡过程，不同的药物可以通过不同的信号途径诱导细胞凋亡。     

染料木黄酮诱导人结肠癌SW480细胞凋亡及机制

目的 探讨染料木黄酮（GEN）诱导人结肠癌SW480细胞凋亡的生物学效应及抗肿瘤机制。方法 通过MTT法检测GEN对该细胞系生长的影响；应用PI染色流式细胞术（FCM）分析GEN处理前后的细胞周期分布的变化；SP免疫组化法检测细胞内bcl-2、Bax蛋白的表达。结果 GEN能抑制SW480细胞增殖，呈浓度依赖性。流式细胞仪示不同浓度GEN作用于SW480细胞，出现G2／M阻滞；SP免疫组化结果表明GEN处理细胞72h后，Bax蛋白表达升高，bcl-2蛋白表达下降。结论 GEN有诱导人结肠癌SW480细胞凋亡的作用，其机制与bcl-2／Bax比值下降有关。     

Tripartite meeting in gene and cell therapy, 2008: Irish Society for Gene and Cell Therapy, British Society for Gene Therapy, and International Society for Cell and Gene Therapy of Cancer

The second annual meeting of the Irish Society for Gene and Cell Therapy was held in Cork, Ireland on May 15 and 16, 2008 (http://crr.ucc.ie/isgct/). The meeting was jointly organized with the British Society for Gene Therapy and the International Society for Cell and Gene Therapy of Cancer. Because of the location of the conference and the co-organization of this meeting with the British and International Gene Therapy societies, the meeting enjoyed a range of talks from some of the major leaders in the field. Particularly notable were the talented molecular and cell biologists from Ireland who have contributed cutting edge science to the field of gene therapy. Topics including cardiovascular disease, repair of single-gene disorders, and cancer gene therapy were discussed with presentations ranging from basic research to translation into the clinic. Here we describe some of the most exciting presentations and their potential impact on imminent clinical gene therapy trials.     

骨髓增生异常综合征患者骨髓CD34+细胞 Fas,FasL及Bcl-2的表达和凋亡

为了观察骨髓增生异常综合征 (MDS)患者骨髓CD34+ 细胞Fas ,FasL和Bcl 2的表达和凋亡情况并探讨这些抗原的表达和细胞凋亡的关系。采用流式细胞术测定了 2 6例MDS和 10例急性髓系白血病 (AML)患者及 6例非血液病患者 (对照 )骨髓CD34+ 细胞的Fas,FasL和Bcl 2表达率和细胞凋亡率。结果显示 ,各型MDS患者CD34+ 细胞Fas和FasL表达率较对照组明显增加 (P <0 .0 1) ,Bcl 2的表达率除难治性贫血 /环形铁粒幼细胞性难治性贫血 (RA/RAS)与对照组无显著差异 (P >0 .0 5 )外 ,难治性贫血伴有原始细胞增多 (RAEB)和转变中的难治性贫血伴原始细胞增多 (RAEB t)患者均明显高于对照组 (P <0 .0 1) ;各型MDS间CD34+ 细胞Fas的表达率相近 ,而Bcl 2的表达率却存在非常显著性差异 ,即RA/RAS 相似文献   

新辅助化疗诱导结肠癌细胞凋亡及IκBα mRNA的表达

目的探讨羟基喜树碱 (HCPT)联合 5 氟尿嘧啶 (5 Fu)和甲酰四氢叶酸钙 (CF)新辅助化疗诱导结肠癌细胞凋亡及其可能的分子机制。方法2 0例结肠癌患者术前静脉滴入HCPT 10mg/d ,5 Fu5 0 0mg/d,CF 2 0 0mg/d ,共 5天即一周期。术中取结肠癌组织标本 ,应用DNA琼脂糖凝胶电泳检测细胞凋亡的生化改变 ,比色法检测Caspase 3的活性 ,RT PCR半定量技术检测IκBαmRNA的表达 ,并与同期术中取得的 2 0例术前未化疗患者的结肠癌组织及癌旁正常肠组织作比较。结果①新辅助化疗组结肠癌细胞DNA琼脂糖凝胶电泳出现明显的特征性梯状条带 ,术前未化疗组结肠癌细胞无明显的DNA梯状条带 ;②结肠癌细胞Caspase 3活性新辅助化疗组较术前未化疗组结肠癌细胞及癌旁正常结肠细胞明显增高 (P <0 .0 5 ) ;③新辅助化疗诱导后 ,结肠癌细胞IκBαmRNA的表达明显上调 (P <0 .0 5 )。结论HFCF方案新辅助化疗可能通过上调IκBαmRNA表达、激活Caspase 3活性诱导人体内结肠癌细胞凋亡而起作用。     

消化道肿瘤组织芯片中BAG-1的表达及意义分析

目的 检测抗凋亡基因BAG-1在多种消化道肿瘤中的表达，并讨论其意义。方法 利用组织芯片，采用免疫组化DAKO Envision法检测食道癌、胃癌、直肠癌及正常组织中BAG-1的表达。结果 食道癌、胃癌及直肠癌组织中BAG-1的表达比其对应的正常组织为高（P〈0．01）。结论 BAG-1在消化道肿瘤组织中表达高于正常组织，提示BAG-1表达可能与消化道恶性肿瘤的发生有关。     

X线、CT联合骨密度测量在股骨头无菌坏死中的临床应用

目的:探讨X线、CT结合骨密度测量对股骨头无菌坏死(FHN)的程度及治疗效果的评估价值.方法:对32例41个FHN在治疗前、后3个月进行髋关节X线平片检查、CT检查和股骨头的骨密度测量,比较FHN治疗前后的骨密度及影像学的变化.结果:41个坏死的股骨头由X线确诊病灶24个,由CT确诊病灶17个;异常股骨头的骨密度值(BMD)和骨矿含量(BMC)与正常股骨头对比差异明显(P <0.05),FHN治疗前后BMD及BMC比较差异显著(P <0.05).结论:运用X线、CT联合骨密度仪测量骨密度值对FHN的程度及治疗效果具有较好的评估价值.     

62例放疗与化疗后骨髓抑制的治疗及护理

1999年 1月— 2 0 0 3年 1月 ,我们采用参附注射液或吉赛欣或二者合用治疗放疗与化疗后骨髓抑制 62例 ,取得了良好的效果 ,现报告如下。1　临床资料选择我院收治的各种恶性疾病放疗、化疗后及少数其他疾病治疗后骨髓抑制病人 62例 ,其中急性白血病 16例 ,恶性淋巴瘤 9例 ,多发性骨髓瘤 12例 ,慢粒急变或加速期 8例 ,肺癌 9例 ,食道癌 6例 ,牛皮癣 1例 ,甲状腺功能亢进 1例 ;年龄 4岁～69岁 ,平均年龄 36.5岁 ;男 36例 ,女 2 6例。2　治疗方法62例随机分为A组、B组、C组 ,A组单用吉赛欣 (华北制药集团金坦生物技术开发有限公司 ) 75 μg皮…     

鼠尾草酸对NB4细胞生长及凋亡、分化作用的体外研究

目的 观察鼠尾草酸（Carnosic acid,CA）对NB4细胞生长及凋亡、分化的诱导作用.方法 通过四氮唑蓝比色（MTT）,细胞形态,流式细胞仪测定细胞周期、凋亡率及CD14的表达,观察鼠尾草酸对NB4细胞的影响.结果 NB4细胞经2.5μmol/L以上浓度的鼠尾草酸作用后增殖受到抑制,2.5μmol/L、5μmol/L、10μmol/L CA 作用72h后,细胞形态呈现凋亡细胞的特征.2.5μmol/L、5μmol/L、10μmol/L CA 作用于NB4细胞72h凋亡率分别为7.216%、11.131 %、20.336%,与对照组相比差异显著,G0/G1期细胞阻滞.CD14的表达与对照组相比无差异性.结论 CA对NB4细胞有生长抑制作用,能诱导NB4细胞发生凋亡,具有一定的量效和时效关系,单独使用CA的分化作用不显著.     

Effects of Androgen Deprivation Therapy on Pain Perception,Quality of Life,and Depression in Men With Prostate Cancer

Context

Previous animal and human research suggests that testosterone has antinociceptive properties. Castration in male rodents increases pain perception which is reversed by testosterone replacement. Pain perception also improves in hypogonadal men with testosterone therapy. However, it remains unclear whether androgen deprivation therapy (ADT) in men with prostate cancer (PCa) is associated with an increase in pain perception.

Mapping Bone Marrow Response in the Vertebral Column by Positron Emission Tomography Following Radiotherapy and Erlotinib Therapy of Lung Cancer

Molecular Imaging and Biology - To map functional bone marrow (BM) by 2-deoxy-2-[18F]fluoro-d-glucose ([18F]FDG) positron emission tomography (PET) in the vertebral column of lung cancer patients...     

First-in-man Phase 1 Clinical Trial of Gene Therapy for Advanced Pancreatic Cancer: Safety,Biodistribution, and Preliminary Clinical Findings

This phase 1 trial was aimed to determine the safety, pharmacokinetics, and preliminary clinical activity of CYL-02, a nonviral gene therapy product that sensitizes pancreatic cancer cells to chemotherapy. CYL-02 was administrated using endoscopic ultrasound in 22 patients with pancreatic cancer that concomitantly received chemotherapy (gemcitabine). The maximum-tolerated dose (MTD) exceeded the maximal feasible dose of CYL-02 and was not identified. Treatment-related toxicities were mild, without serious adverse events. Pharmacokinetic analysis revealed a dose-dependent increase in CYL-02 DNA exposure in blood and tumors, while therapeutic RNAs were detected in tumors. No objective response was observed, but nine patients showed stable disease up to 6 months following treatment and two of these patients experienced long-term survival. Panels of plasmatic microRNAs and proteins were identified as predictive of gene therapy efficacy. We demonstrate that CYL-02 nonviral gene therapy has a favorable safety profile and is well tolerated in patients. We characterize CYL-02 biodistribution and demonstrate therapeutic gene expression in tumors. Treated patients experienced stability of disease and predictive biomarkers of response to treatment were identified. These promising results warrant further evaluation in phase 2 clinical trial.