晚期恶性肿瘤患者临床漏 /误诊分析 - - 附 35例尸解报告

目的：在诊断方法、技术不断进步的情况下,确定恶性肿瘤临床与尸检 诊断的不一致率。方法：对本院肿瘤科5年来（1994－1999年）所有尸检进行回顾性研究。结果：共尸检35例,男性24例,女性11例,中位年龄64岁（29－84岁）。35例被尸解的尸体中共发现41个恶性肿瘤,其中10例、16个恶性肿瘤被误诊或漏诊。该10例中6例死亡原因为恶性肿瘤。在本研究中,临床与尸检诊断的不一致率为39％。结论：恶性肿瘤临床与尸检诊断的不一致率依然很高,本结果证实了尸检的重要性。     

人NGAL克隆表达及抗体的制备与鉴定

目的：高效表达人中性粒细胞明胶酶相关脂质运载蛋白（neutrophil gelatinase-associated lipocalin,NGAL）重组蛋白并制备其多克隆及单克隆抗体。方法：人工合成经密码子优化的NGAL基因,构建原核表达重组质粒pW28-NGAL,IPTG诱导表达后分离纯化并分析在溶液中的聚集状态;获得的rhNGAL抗原免疫兔子制备多克隆抗体,免疫小鼠筛选高效价的特异性单克隆抗体并鉴定抗体亚型;Protein G亲合柱纯化,等温滴定量热法及非竞争ELISA法检测抗体的亲和力,SDS-PAGE分析纯度;最后运用Western blot、免疫荧光和免疫组化对抗体进行鉴定。结果：高效获得高纯度NGAL重组蛋白,主要以单体形式存在;成功制备兔多抗,同时筛选获得6株高效价单克隆抗体;其中,25号单抗为IgG1亚型,余者均属IgG2a亚型,且19、35号单抗的亲和常数分别为3.06×109、2.14×109。SDS-PAGE分析表明纯度均大于90%。进一步的鉴定结果表明制备的抗体皆具有良好免疫反应性及特异性,且单抗的特异性明显高于多抗。结论：本研究利用密码子优化技术高效表达制备了NGAL重组蛋白,获得了其高效价多克隆抗体和高特异性单克隆抗体,为NGAL的病理生理作用等功能研究提供了良好的实验材料,为NGAL临床免疫检测诊断试剂的国产化奠定了有力的基础。     

新疆地区维、汉民族妇女散发性乳腺癌BARD1基因突变检测分析

背景与目的:乳腺癌是基因环境交互作用的复杂疾病,高共显的易感基因突变与家族性乳腺癌密切相关.本研究旨在探讨新疆地区维、汉民族女性散发性乳腺癌患者BARD1基因突变情况及突变位置,以期为后续研究奠定基础.方法:随机选取维、汉民族妇女散发性乳腺癌各30例,收集临床病理资料及癌组织与血液标本.应用启动子和外显子测序的方法检测占BARD1基因突变,并在血液样本中测序来验证上述结果.结果:(L)维族和汉族散发性乳腺癌患者在生育史上差异有统计学意义,且维族有很大比例为非雌激素依赖性乳腺癌.(2)维族散发性乳腺癌患者BARD1基因突变率为33.3%,而汉族为13.8%,但差异无统计学意义(X<'2>=3.11,P=0.07).新发现的14个低频SNP全部位于非编码区.(3)血液验证除一个位点rs28997575外,其余43个全部一致,提示突变为生殖细胞突变.结论:维、汉民族的乳腺癌类型有差异,治疗也应区别对待.BARD1基因突变在维族乳腺癌患者中作用可能大于汉族患者,机制可能是增加了乳腺癌易感性.     

Identifying dysfunctional miRNA-mRNA regulatory modules by inverse activation,cofunction, and high interconnection of target genes: A case study of glioblastoma

Background

Accumulating evidence demonstrates that complex diseases may arise from cooperative effects of multiple dysfunctional miRNAs. Thus, identifying abnormal functions cooperatively regulated by multiple miRNAs is useful for understanding the pathogenesis of complex diseases.

Methods

In this study, we proposed a multistep method to identify dysfunctional miRNA-mRNA regulatory modules (dMiMRMs) in a specific disease, in which a group of miRNAs cooperatively regulate a group of target genes involved in a specific function. We identified dysfunctional miRNAs, which were differentially expressed and inversely regulated most of their target genes, by integrating paired miRNA and mRNA expression profiles and miRNA target information. Then, we identified cooperative functional units, in each of which a pair of miRNAs cooperatively repressed function-enriched and highly interconnected target genes. Finally, the cooperative functional units were assembled into dMiMRMs.

Results

We applied our method to glioblastoma (GBM) and identified GBM-associated dMiMRMs at the population, subtype, and individual levels. We identified 5 common dMiMRMs using all GBM samples, 3 of which were associated with the prognosis in patients with GBM and were better predictors of prognosis than were miRNAs or mRNAs alone. By applying our approach to GBM subtypes, we found consistent dMiMRMs across GBM subtypes, and some subtype-specific dMiMRMs were observed. Furthermore, personalized dMiMRMs were identified, suggesting significant individual differences in different patients with GBM.

Conclusions

Our method provides the capability to identify miRNA-mediated dysfunctional mechanisms underlying complex diseases.     

荧光定量聚合酶链反应技术在胃幽门螺杆菌检测中的应用

目的:评估荧光定量PCR在检测胃粘膜上幽门螺杆菌的DNA中的应用价值。方法:用荧光定量PCR与普通PCR检测胃粘膜上的幽门螺杆菌的DNA,并对二者进行方法学比较。结果:荧光定量PCR阳性率为95.3%(143/150),普通PCR阳性率仅为86.0%(129/150),二者之间有明显的差异(P<0.01)。结论:荧光定量PCR在临床上检测幽门螺杆菌DNA方面具有简单、快速、敏感、特异的特点,在临床上具有较高的应用价值。     

NEDD9在食管鳞状细胞癌中表达的意义

目的:探讨NEDD9在食管鳞癌中表达的意义及与肿瘤发生发展和转移的关系。方法:采用免疫组化方法及Real-time PCR技术检测NEDD9在40例食管鳞癌癌组织、癌旁正常组织中的表达情况,并分析NEDD9的表达与肿瘤临床病理特征之间的关系。结果:食管癌组织与癌旁正常组织中NEDD9蛋白阳性表达率分别为77.5%、25.0%,mRNA表达CT值分别为1.704±0.360、0.843±0.526,食管癌组织中NEDD9蛋白及mRNA水平均明显高于癌旁组织及正常食管组织。NEDD9的表达与患者年龄、性别无关(P>0.05),与肿瘤组织分化程度、TNM分期及有无淋巴结转移有关(P<0.05)。结论:NEDD9在食管癌组织中表达明显高于正常癌旁食管组织,可能促进食管癌的侵袭及转移,可作为食管癌治疗的潜在靶点。     

强化院内感染管理对肿瘤术后患者控制院内感染的影响研究

目的 探讨强化院内感染管理对肿瘤术后患者控制院内感染的影响.方法 回顾性分析行手术治疗的80例肿瘤患者的临床资料,根据不同的护理方法将其分为研究组(n=42)和对照组(n=38).对照组患者接受常规管理干预,研究组患者接受强化院内感染管理干预.护理后,比较两组患者的院内感染率,比较两组患者的护理规范、护理效率、人文关怀、医德医风4个护理指标的质量评分情况,记录患者的住院时间、住院费用和患者对健康知识的掌握情况,并比较两组患者对治疗方案的满意度.结果 护理后,研究组患者的院内感染率为4.8％,明显低于对照组患者的29.0％,差异有统计学意义(P<0.01);护理后,研究组患者对护理规范、护理效率、人文关怀、医德医风指标的质量评分均明显高于对照组患者,差异均有统计学意义(P<0.01);研究组患者的住院时间明显短于对照组,住院费用明显少于对照组,对健康知识的掌握率明显高于对照组患者(P<0.01).护理后,研究组患者的满意度为95.2％,明显高于对照组患者的79.0％,差异有统计学意义(P<0.01).结论 强化院内感染管理可降低肿瘤术后患者的院内感染率,缩短患者的住院时间,提高护理质量以及患者的满意度,临床效果显著.     

Bone‐induced c‐kit expression in prostate cancer: A driver of intraosseous tumor growth

Loss of BRCA2 function stimulates prostate cancer (PCa) cell invasion and is associated with more aggressive and metastatic tumors in PCa patients. Concurrently, the receptor tyrosine kinase c‐kit is highly expressed in skeletal metastases of PCa patients and induced in PCa cells placed into the bone microenvironment in experimental models. However, the precise requirement of c‐kit for intraosseous growth of PCa and its relation to BRCA2 expression remain unexplored. Here, we show that c‐kit expression promotes migration and invasion of PCa cells. Alongside, we found that c‐kit expression in PCa cells parallels BRCA2 downregulation. Gene rescue experiments with human BRCA2 transgene in c‐kit‐transfected PCa cells resulted in reduction of c‐kit protein expression and migration and invasion, suggesting a functional significance of BRCA2 downregulation by c‐kit. The inverse association between c‐kit and BRCA2 gene expressions in PCa cells was confirmed using laser capture microdissection in experimental intraosseous tumors and bone metastases of PCa patients. Inhibition of bone‐induced c‐kit expression in PCa cells transduced with lentiviral short hairpin RNA reduced intraosseous tumor incidence and growth. Overall, our results provide evidence of a novel pathway that links bone‐induced c‐kit expression in PCa cells to BRCA2 downregulation and supports bone metastasis.     

沙利度胺对人非霍奇金淋巴瘤Raji细胞VEGF影响的研究

目的 探讨沙利度胺体外对人类非霍奇金淋巴瘤(NHL)Raji细胞血管内皮生长因子(VEGF)的影响及其作用机制.方法 采用MTT法检测沙利度胺对人类NHL Raji细胞是否有抑制作用,采用ELISA法检测沙利度胺作用前后Raji细胞上清液中VEGF含量的变化.结果 MTT法示沙利度胺体外对人类NHL Raji细胞无直接的抑制作用(P＞0.05).ELISA法示10、25、50 mg/L的沙利度胺组对Raji细胞分泌VEGF均有明显抑制作用(P＜0.05).5 mg/L沙利度胺组及0.1%DMSO对照组对Raji细胞分泌VEGF无明显抑制作用(P＞0.05).沙利度胺对Raji细胞分泌VEGF的抑制作用随着浓度和时间的递增而增加.结论沙利度胺能够抑制人类NHL Raji细胞分泌VEGF并且存在时间和浓度依赖性.沙利度胺体外对Raji细胞并无明显直接的抗增生效应与细胞毒活性.