Pilot study of an HLA-A2 peptide vaccine using flt3 ligand as a systemic vaccine adjuvant

A pilot vaccine study was conducted to test the safety and immunological efficacy of four monthly immunizations of an MHC class I peptide vaccine, the E75 HLA-A2 epitope from HER-2/neu, using flt3 ligand as a systemic vaccine adjuvant. Twenty HLA-A2-expressing subjects with advanced stage prostate cancer were randomly assigned to one of four immunization or treatment schedules: (a) Flt3 ligand (20 g/kg per day) administered subcutaneously daily for 14 days on a 28-day cycle, monthly for four months; (b) flt3 ligand course as above with the E75 peptide vaccine administered on day 7 of each flt3 ligand cycle; (c) flt3 ligand course as above with the E75 peptide vaccine administered on day 14 of each flt3 ligand cycle; or (d) E75 peptide admixed with granulocyte–macrophage colony-stimulating factor and administered intradermally once every 28 days, as has previously been reported. The primary endpoints of the study were the determination of safety and immunological efficacy in generating E75-specific T cells as determined by peptide-specific interferon-gamma ELIspot. Adverse events included one grade 3 skin reaction and the development of grade 2 autoimmune hypothyroidism in two subjects with preexisting subclinical autoimmune hypothyroidism. Dendritic cells were markedly increased in the peripheral blood of subjects receiving flt3 ligand with each repetitive cycle, but augmentation of antigen-presenting cells within the dermis was not observed. Apart from a single subject, no significant peptide-specific T-cell responses were detected by ELIspot, whereas delayed-type hypersensitivity responses were detectable in control subjects and in subjects receiving peptide vaccine early in the course of flt3 ligand administration. The absence of robust peripheral immune responses in the current study may be attributable to the small numbers of subjects or differences in the subject population. In addition, the inability of flt3 ligand to augment the number of peripheral skin antigen-presenting cells may have contributed to the absence of robust peptide-specific immunity detectable in the peripheral blood of immunized subjects treated with flt3 ligand.     

Role of intracellular Ca2+ stores in smooth muscle contractions of the guinea pig vas deferens

Summary Guinea pig vas deferens was used as an animal model for alpha-1 adrenoceptor (₁-receptor) mediated contractions in human hyperplastic prostatic tissue. The selective ₁-receptor agonist, phenylephrine (PE), induced fully reversible, dose-dependent contractions antagonized by increasing concentrations of the ₁-receptor blockers prazosin (1–100 nM) and YM 617 (0.1–10 nM). Removal of extracellular Ca²⁺ reduced PE-evoked contractions in a time-dependent manner. Nifedipine (1–1000 nM), a blocker of voltage-dependent L-type Ca²⁺ channels (VDCC), inhibited the PE-induced response by up to 65%. Removal of extracellular Ca²⁺ abolished the ₁-agonist reactivity in a time-dependent fashion. To elucidate the participation of intracellular Ca²⁺ stores in ₁-receptor contractions, the tissue was pretreated with ryanodine (10 M) or thapsigargin (0.1 M), established inhibitors of Ca²⁺ release from intracellular pools. Both substances reduced the PE contractions by up to 80%. Nifedipine suppressed the remaining contractions completely. This provides evidence that Ca²⁺ influx through VDCC and Ca²⁺ release from intracellular stores contribute to ₁-receptor contractions in the guinea pig vas deferens and may be important in obstructive benign prostatic hyperplasia.     

分析前列腺特异抗原检测结果预测前列腺癌发病趋势

目的　回顾我院 4年来前列腺疾病患者血清前列腺特异抗原 (PSA)测定结果与前列腺癌的检出率 ,分析兰州地区前列腺癌的发病趋势。方法　收集了 165 8例前列腺疾病患者血清PSA结果 ,城镇职工792例 ,农民 866例。对其中 2 37例PSA水平 >10 μg·L- 1者行前列腺穿刺活检。结果　检出前列腺癌 35例 ,占总例数 2 .1% ,占活检病例 14 .7% ;前列腺癌组血清PSA水平高于非前列腺癌组 (P <0 .0 1) ;城镇职工前列腺癌发病率逐年升高。结论　PSA是筛查前列腺癌的重要方法 ,兰州地区前列腺癌发病率逐年上升趋势 ,城镇职工发病率升高显著。     

血清游离/总前列腺特异性抗原比值在前列腺癌诊断中的应用

目的:探讨血清游离态PSA(F-PSA)和总PSA(T-PSA)及游离态/总(F/T)PSA比值作为前列腺疾病诊断指标的价值。方法:采用Abbott公司AxSYM化学发光免疫分析仪自动检测未经治疗的84例前列腺增生(BPH)病人和29例前列腺癌病人血清F-PSA和T-PSA,并计算F/T比值。结果:F/T百分率值在前列腺癌组中明显低于前列腺增生组(P<0.001)。若总PSA限定于4～10μg/L范围内,应用F/T百分率值可区别前列腺癌与前列腺增生(P<0.01),而两组总PSA差异无显著意义(P>0.05)。结论:F/T百分率值可用于区别前列腺癌与前列腺增生;总PSA在4～10μg/L范围时,应用F/T百分率值较总PSA为佳。     

棕榈酰化蛋白质组学分析揭示前列腺癌细胞中雄激素促进代谢相关蛋白棕榈酰化修饰

目的 筛选棕榈酰化修饰水平受雄激素诱导的蛋白,探索前列腺癌去雄激素治疗外其他潜在的治疗靶点.方法: 以LNCaP细胞为研究对象,雄激素(Methyltrienolone,R1881,5 nmol/L)或DMSO(dimethyl sulfoxide)处理LNCaP细胞24 h,同时利用人工合成的炔基棕榈酸Alk-C16 (100 μmol/L)对细胞进行代谢标记,收集细胞,裂解,提取总蛋白,加入标记有叠氮化物的琼脂糖珠 (1 mmol/L), 室温反应1 h,利用叠氮化物与Alk-C16末端炔基发生点击化学反应形成的共价键将棕榈酰化修饰蛋白富集在琼脂糖珠上,进行蛋白质谱非标记定量分析(label-free quantitation, LFQ), 比较R1881处理和非处理细胞蛋白棕榈酰化修饰变化情况,筛选棕榈酰化修饰水平受雄激素诱导的蛋白.结果: 实验共鉴定出907个潜在的棕榈酰化修饰蛋白(mascot score> 2, P<0.05), 其中有430个蛋白LFQ值至少有2次不为0.在这430个蛋白中有92个蛋白R1881处理与非处理样品LFQ比值大于 1.5(P<0.05), 说明雄激素能够显著促进该蛋白棕榈酰化修饰.利用Cytoscape软件对92个蛋白进行功能富集分类,发现已知功能蛋白可分为代谢相关,蛋白折叠相关和翻译起始相关3类,其中,代谢相关蛋白包括脂代谢(6个),糖代谢(7个)和呼吸电子传递链(8个)3部分,另外还有少量氨基酸代谢(2个)和其他代谢相关蛋白(2个).参与呼吸电子传递链的细胞色素b-c1复合体亚基2 (cytochrome b-cl complex subunit2, UQCRC2) 雄激素R1881处理和未处理样品LFQ比值最高(>3,P<0.05),说明该蛋白棕榈酰化修饰受雄激素诱导最为明显.LFQ比值最高为UQCRC2,其次为脂代谢相关的长链特异性酰基辅酶A脱氢酶(very long-chain specific acyl-CoA dehydrogenase, ACADVL)和糖代谢相关的6-磷酸葡萄糖酸脱氢酶(6-phosphogluconate dehydrogenase, PGD), 但其LFQ比值均未超过3.结论: 代谢尤其是呼吸电子传递链相关蛋白的棕榈酰化调控机制的研究可能将为前列腺癌的诊疗和靶向药的研发提供新的指导思路.     

抑那通在前列腺癌内分泌治疗中的临床意义

目的为提高晚期前列腺癌内分泌治疗效果,１９９０年７月～１９９２年７月间对１７例前列腺癌患者进行了抑那通（药物除睾）的临床观察。方法随访３～５年获得可评价资料１６例病例,采用免疫酶联法测定ＰＳＡ变化,放免法检测性激素水平,用Ｉ－ＰＳＳ表调查排尿及生活质量,对前列腺及转移病灶、全身状况的变化进行对比观察。结果抑那通治疗后睾酮降至去势水平（Ｐ＜０．０１）,前列腺特异抗原及其他肿瘤标记物提示反应性良好,症状及全身状况明显改善。结论每月注射一次抑那通,如果与抗男性素剂联合使用可获得更安全有效的治疗效果。     

Phase II trial of oral 1,25-dihydroxyvitamin D (calcitriol) in hormone refractory prostate cancer

Epidemiologic and experimental data support a role for 1,25-dihydroxyvitamin D₃ in the growth regulation of prostate cancer. We conducted a phase II clinical trial evaluating calcitriol (1,25(OH)₂D₃) in patients with hormone refractory prostate cancer. We enrolled 14 patients in this study. 1,25(OH)₂D₃ was initiated at a daily oral dose of 0.5 μg and escalated to 1.5 μg daily. No objective responses were observed. However, in two patients decreases of 25% and 45% in prostate specific antigen levels were seen. Hypercalcemia was the predominant toxicity. We conclude that 1,25(OH)₂D₃ given in this manner is inactive in advanced prostate cancer. Dose escalation of oral 1,25(OH)₂D₃ is limited by hypercalcemia.     

顾乃强乳腺增生病电脑诊疗专家系统的医理设计——附130例临床验证

此专家系统在修善顾博士行医经验和治疗原则的基础上 ,利用多媒体技术 ,结合声频、视频、图片和文本以治疗乳腺增生病。在医疗设计中 ,着重于顾博士的类型识别要点和乳腺增生病的自然规律。尤其 ,此医疗设计阐明了顾博士所总结的有价值的治疗经验 ,以突出此专家系统的科学性、实用性和可操作性。笔者曾将此专家系统应用于临床 1 30份病例的治疗中。其结果表明此专家系统的临床治疗正确率与顾博士临床治疗相对照达 98%。     

良性增生前列腺组织中鸟氨酸脱羧酶基因表达的研究

为探讨鸟氨酸脱羧酶（ＯＤＣ）基因表达与前列腺良性增生的关系,采用Ｎｏｒｔｈｅｒｎ印迹和ＲＮＡ斑点杂交技术,测定分析了正常人（１０例）和良性前列腺增生症（ＢＰＨ）病人（１５例）前列腺组织中ＯＤＣｍＲＮＡ的丰度。结果显示,良性增生前列腺组织中ＯＤＣｍＲＮＡ丰度较正常前列腺组织显著升高。经密度扫描测定结果显示,Ｎｏｒｔｈｅｒｎ印迹分析中ＢＰＨ前列腺组织中ＯＤＣｍＲＮＡ丰度约为对照组的６～２０倍,斑点杂交为５倍以上。提示ＯＤＣ基因表达增强在良性前列腺增生症形成中起重要作用