PRNCR1在前列腺癌诊断及预后的价值

目的 探讨前列腺癌非编码RNA1（PRNCR1）在前列腺癌早期诊断及预后的价值。方法 选取2012年1月至2015年10月在本院诊治前列腺癌患者45例、良性前列腺增生患者88例，进行PRNCR1检验。结果 PRNCR1 检验的灵敏度84.4%（38/45）、特异度90.9%（80/88）、阳性预测值82.6%（38/46）、阴性预测值92.0%（80/87)、符合率88.7%（118/133），PRNCR1在前列腺癌组织中含量高于癌旁组织和良性前列腺增生组织，差异有统计学意义（P<0.05）。两年内转移、复发、肿瘤死亡患者30例，其PRNCRA表达量高于两年内无转移、复发、死亡的患者（15例），差异有统计学意义（P<0.05）。结论 PRNCR1表达在诊断前列腺癌有价值，PRNCR1高表达与患者预后不良有关。     

Inactivated influenza vaccine formulated with single-stranded RNA-based adjuvant confers mucosal immunity and cross-protection against influenza virus infection

Influenza vaccination is considered the most valuable means to prevent and control seasonal influenza infections, which causes various clinical symptoms, ranging from mild cough and fever to even death. Among various influenza vaccine types, the inactivated subunit type is known to provide improved safety with reduced reactogenicity. However, there are some drawbacks associated with inactivated subunit type vaccines, with the main ones being its low immunogenicity and the induction of Th2-biased immune responses. In this study, we investigated the role of a single-stranded RNA (ssRNA) derived from the intergenic region in the internal ribosome entry site of the Cricket paralysis virus as an adjuvant rather than the universal vaccine for a seasonal inactivated subunit influenza vaccine. The ssRNA adjuvant stimulated not only well-balanced cellular (indicated by IgG2a, IFN-γ, IL-2, and TNF-α) and humoral (indicated by IgG1 and IL-4) immune responses but also a mucosal immune response (indicated by IgA), a key protector against respiratory virus infections. It also increases the HI titer, the surrogate marker of influenza vaccine efficacy. Furthermore, ssRNA adjuvant confers cross-protective immune responses against heterologous influenza virus infection while promoting enhanced viral clearance. Moreover, ssRNA adjuvant increases the number of memory CD4⁺ and CD8⁺ T cells, which can be expected to induce long-term immune responses. Therefore, this ssRNA-adjuvanted seasonal inactivated subunit influenza vaccine might be the best influenza vaccine generating robust humoral and cellular immune responses and conferring cross-protective and long-term immunity.     

人端粒酶RNA的cDNA探针制备及其对胃粘膜细胞端粒酶RNA表达的检测

目的 建立人端粒酶RNA表达的检测方法。方法 制备人端粒酶RNA，(human telomeraseRNA，hTR)的cDNA探针，分别应用RNA斑点杂交与端粒重复序列扩增法(TRAP)分析检测不同胃粘膜的端粒酶RNA的表达与端粒酶活性。结果 人端粒酶RNA的cDNA探针制备成功。18例活检胃癌组织及45例手术胃癌组织RNA斑点杂交检测的阳性率均为l00％，相应TRAP分析的阳性率分别为88．89％、86．67％，低于RNA斑点杂交(P＜0．05)。同时RNA斑点杂交结果提示在非癌胃组织中随着肠化程度增高人端粒酶RNA表达也增强。结论 RNA斑点杂交检测人端粒酶RNA，具有高度的敏感性和特异性，弥补了TRAP分析敏感性不足的缺点。     

外周血细胞DNA、RNA及血红蛋白同时提取法

采用淋巴细胞分离液分离外周血中的有核细胞,经细胞裂解液处理,上清提取RNA,沉淀提取DNA,用分离液分离所得红细胞沉淀制备血红蛋白溶液。经对抽提物质量进行评价,发现3种提取物质量均高。本法能从少量外周血中同时分离DNA、RNA及血红蛋白,高效易操作,值得推广。     

慢性低氧大鼠肺动脉内皮素mRNA的表达及定位

目的：探讨慢性低氧大鼠肺动脉内皮素－1基因表达及分布。方法：采用生物素标记cRNA探针，对Wistar大鼠肺动脉进行原位杂交。结果：低氧1周组及低氧2周组大鼠大部分肺动脉内皮细胞ET－1mRNA表达呈阳性信号，而对照组大鼠仅极少数阳性信号（P＜0．01）；低氧1周组大鼠部分显示强阳性信号。低氧1周组及低氧2周组大鼠部分肺动脉平滑肌细胞呈现ET-1mRNA表达阳性信号，而对照组大鼠则无阳性信号表达（P＜0．01及0．05）。结论：慢性低氧对大鼠肺动脉ET－1分泌的影响是在基因转录水平进行，且主要细胞定位在肺动脉内皮细胞，此外还包括肺动脉平滑肌细胞。     

核酸及核苷酸用于营养及免疫调节治疗的早期卫生技术评估

目的 评价核酸或核苷酸用于临床营养支持和免疫调节治疗的安全性、有效性和经济学价值。方法 检索中国生物医学文献数据库、Cochrane图书馆、MEDLINE光盘数据库、EMBASE光盘数据库,并联机检索SCI数据库,鉴定有关随机对照试验(RCTs),采用RevMan4.l进行Meta-分析。结果 纳入46个随机对照试验,涉及核酸/核苷酸用于肠内营养支持、婴儿营养、免疫调节治疗。18个研究报告了含核苷酸组分的免疫增强型肠内营养对外科术后和危重症患者预后的影响,合并分析发现免疫营养对患者感染率、住院时间和费用有肯定意义。发现1个报告在母乳代用品中加入核苷酸对婴儿免疫功能影响的研究,但报告对临床结局无明显影响。27个对“免疫核酸”用于免疫调节治疗的随机对照试验均为低质量研究,合并分析无法肯定iRNA的临床价值。结论 在外科术后患者应用免疫肠内营养制剂可降低感染率、缩短住院时间并可能减少住院费用,但不能确定作为组分之一的核苷酸的作用。尚无证据支持在母乳代用品中添加核苷酸具有临床意义。不能肯定免疫核酸是否具有免疫调节作用,亦未发现核酸在延缓衰老、改善老年人健康状况方面的可用证据。建议对核酸、核苷酸用于营养支持治疗和免疫调节治疗进行进一步研究,严格规范“核酸营养品”的宣传和应用。     

Differential Regulation of RGS-2 by Constant and Oscillating PTH Concentrations

PTH has diverse effects on bone metabolism: anabolic when given intermittently, catabolic when given continuously. The cellular mechanisms underlying the varying target cell response are not clear yet. PTH induces RGS-2, a member of the Regulator of G-protein Signaling protein family, via cAMP/PKA, and inactivates PKC-mediated signaling. To investigate intracellular signaling pathways with different PTH concentration-time patterns, we treated UMR 106-01 osteoblast-like cells in a perfusion system. PTH was administered intermittently (4 min/h, 10⁻⁷ M) or continuously at an equivalent cumulative dose (6.6 × 10⁻⁹ M). cAMP was measured using radioimmunoassay, mRNA levels using real-time rtPCR and ribonuclease protection assay, and protein levels using Western immunoblotting. A single PTH pulse transiently increased cAMP levels by 2000% ± 1200%. In contrast to continuous PTH exposure, cAMP induction remained unchanged with intermittent PTH, ruling out desensitization of the PTH receptor. In continuously perfused cells, RGS-2 abundance was three to five times higher than in cells intermittently exposed to PTH for up to 12 h. MKP-1 and -3 were significantly less induced with pulsatile PTH; exposure-mode-dependent differences in MMP-13 and IGFBP-5 were small. Pulsatile but not continuous PTH administration prevents PTHrP receptor desensitization and accumulation of RGS-2 in osteoblasts, which should preserve PKC-dependent signaling.