宫颈上皮内瘤变与正常宫颈组织差异表达蛋白分析

目的探讨蛋白质组学方法筛查正常宫颈与宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)组织中的差异表达蛋白质标志物,为研究CIN发生的分子机制及临床诊治工作提供依据。方法收集2008年8月至2009年9月间首都医科大学附属北京妇产医院妇瘤科收治的正常子宫颈组织因子宫肌瘤手术行全子宫切除术患者9例为对照组、CIN组织23例(其中CINⅠ7例、CINⅡ8例、CINⅢ8例)。应用二维荧光差异凝胶电泳(two-dimensional fluorescence difference in gel electrophoresis,2-DDIGE)及DeCyder软件寻找差异表达蛋白质点,基质辅助激光解析飞行时间串联质谱(MALDI-TOF/TOF MS)分析差异蛋白质点,数据库搜索鉴定差异表达最明显的S100蛋白家族A9(S100 calcium-binding protein A9,S100A9),真核延长因子1α1(eukaryoticelongation factor 1-alpha-1,eEF1A1)及丙酮酸激酶M2(pyruvate kinase M2,PKM2)3种蛋白质;应用免疫组织化学法(immunohistochemistry,IHC)(其中正常宫颈组织10例、CINⅠ10例、CINⅡ10例、CINⅢ10例)及蛋白印记法(Western blotting)(其中正常宫颈组织12例,CINⅠ～Ⅱ12例,CINⅢ12例)进一步验证上述3种蛋白在CIN组织和正常宫颈组织的表达差异。结果获得CIN组织与正常宫颈组织2-D DIGE图,成功鉴定25个蛋白;免疫组化及蛋白印迹验证结果提示:S100A9蛋白表达于细胞质中,在CIN中表达水平高于正常组,差异有统计学意义(P=0.010);eEF1A1蛋白表达于细胞质中、PKM2蛋白表达于细胞核中,2者在CIN中表达水平均低于正常组,差异有统计学意义(P分别为0.352,0.000)。结论在正常宫颈组织与CIN组织之间存在差异蛋白质表达,S100A9蛋白可能成为辅助诊断CIN的相关标志物,PKM2蛋白可能成为抑制CIN发生的蛋白质,并可根据2种差异蛋白表达预测CIN的发生发展。     

宫颈薄层液基细胞中P16ink4a的表达在宫颈癌筛查中的意义

目的探讨肿瘤抑制基因P16ink4a的编码产物p16蛋白的表达在宫颈液基薄层细胞学（liquid based cytologi-cal test,LCT）筛查中的意义及作为宫颈上皮内瘤变（cervical intraepithelial neoplasia,CIN）生物标志物的可行性和临床意义。方法选择96例HPV阳性的宫颈液基薄层细胞学剩余标本,制作液基薄片,利用SP免疫组化法检测LCT标本中p16蛋白,并与组织活检结果进行对照。结果 p16蛋白在宫颈细胞学诊断未见上皮内病变及癌变（NILM）、非典型鳞状上皮细胞（ASC）、低度鳞状上皮内病变（LSIL）、高度鳞状上皮内病变（HSIL）的阳性表达率分别为0（0/20）、38.9%（14/36）,25.0%（5/20）、85.0%（17/20）。其中ASC及HSIL组的阳性率显著高于NILM组（P〈0.05）,而LSIL组与NILM组比较差异则无统计学意义（P〉0.05）。与病理组织作对照,病检为慢性宫颈炎的病例中,有1例在LCT上p16蛋白表达阳性（2.70%,1/36）;各级CIN中40例表达阳性（72.7%,40/55）;5例SCC全部表达阳性（100.0%,5/5）,且前两组阳性率差异有统计学意义（P〈0.05）。结论 p16蛋白异常表达与CIN的发生有关,其可作为CIN的标志物,辅助细胞学筛查。     

冠状动脉介入术导致对比剂肾病危险因素分析

目的探讨冠状动脉介入术患者对比剂肾病的危险因素。方法收集我院2008年1月~2009年12月期间住院的172例接受冠状动脉介入术患者的临床资料,比较对比剂使用剂量,测定介入治疗前后血清肌酐,分析发生对比剂肾病危险因素。结果 172例患者发生对比剂肾病32例,发生率18.61%(32/172),肾小球滤过率>60 mL/min患者发生对比剂肾病21例(13%),肾小球滤过率<60 mL/min患者发生对比剂肾病19例(40%)(P<0.001),年龄≥75岁、3支病变、左心室射血分数<40%、血清肌酐>133μmol/L及对比剂剂量>300 mL与对比剂肾病相关(P<0.001)。糖尿病或高血压合并肾功能不全对比剂肾病发病率高于单纯糖尿病或高血压患者(P<0.01)。结论介入治疗术前肾功能受损及其损害程度是术后出现对比剂肾病的最主要危险因素,肾功能不全、年龄≥75岁、多支冠状动脉病变、左心室射血分数<40%、对比剂剂量>300 mL均为对比剂肾病的独立危险因素。糖尿病或高血压病合并肾功能不全可以增加对比剂肾病的临床风险。     

Helicobacter pylori infection is probably the cause of chronic idiopathic neutropenia (CIN)-associated splenomegaly

Splenic volume and Helicobacter pylori (H. pylori) infection were evaluated in 67 patients with chronic idiopathic neutropenia (CIN) and 39 healthy individuals. Using ultrasound, splenomegaly was found in 61.7% of H. pylori-infected subjects compared to only 8.7% noted in the group of H. pylori-non-infected individuals (P < 0.0001). Splenomegaly was also found in 47.8% of CIN patients compared to 12.8% in the group of non-CIN subjects (P = 0.0003). However, applying the two-way ANOVA test, a statistically significant effect on splenic volume was documented for "factor H. pylori " (F1(102) = 16.800, P < 0.0001) but not for "factor CIN" (F1(102) = 3.213, P = 0.0760), suggesting that CIN-associated splenomegaly is probably due to H. pylori infection.     

Soluble c-kit ligand production by bone marrow stromal cells is independent of the degree of neutropenia in patients with chronic idiopathic neutropenia

The levels of soluble c-kit ligand (sKL), also known as Steel factor or stem cell factor, were measured in blood serum and long-term bone marrow culture supernatants of 81 patients with chronic idiopathic neutropenia (CIN) and 22 normal controls using a commercially available enzyme-linked immunosorbent assay (ELISA). We found that the levels of serum and culture supernatant sKL did not differ significantly between patients and control subjects and that both serum and supernatant values of the cytokine did not correlate with the number of circulating neutrophils. Furthermore, we found that the levels of the culture supernatant granulocyte colony-stimulating factor (G-CSF), also measured by ELISA, were significantly increased in the patients compared to controls but individual G-CSF values did not correlate with the values of supernatant sKL. These findings suggest that sKL-producing cells continuously secrete sKL and that cytokine secretion is independent of the degree of neutropenia or the levels of supernatant G-CSF in patients with CIN.     

浅谈宫颈病变诊治中应注意的问题

<正>1宫颈病变1.1广义的宫颈病变包括宫颈炎、宫颈损伤、宫颈肿瘤。1.2狭义的宫颈病变通常指上皮内瘤变(CIN)。CIN是与宫颈浸润癌密切相关的一组癌前病变,它反映了宫颈癌发生发展的连续过程。接近90%[1]CIN有HPV感染,HPV感染和亚临床感染也属于CIN范     

Association of JC virus T-antigen expression with the methylator phenotype in sporadic colorectal cancers

BACKGROUND & AIMS: JC virus (JCV) is a polyomavirus that ubiquitously infects humans and has been implicated in various human cancers. JCV encodes a "transforming" gene, T-antigen (T-Ag), which is believed to mediate the oncogenic potential of the virus. We have previously shown that JCV DNA sequences are usually present in human colorectal cancers (CRCs), and we have provided in vitro evidence that JCV can induce chromosomal instability (CIN) in CRC cells. This study tests the hypothesis that JCV T-Ag expression correlates with one or more forms of genomic or epigenetic instability in sporadic CRCs. METHODS: We characterized 100 sporadic CRCs for microsatellite instability (MSI) and CIN. PCR amplifications were performed for T-Ag sequences, and immunohistochemical (IHC) staining was performed to detect T-Ag expression. De novo methylation of the promoter regions of nine putative tumor suppressor genes thought to play a role in colorectal carcinogenesis was studied by methylation-specific PCR. RESULTS: JCV T-Ag DNA sequences were found in 77% of the CRCs and 56% of these cancers (or 43% of the total) expressed T-Ag by IHC. Significant associations were observed between T-Ag expression and CIN in CRCs (P = .017) and between T-Ag expression and promoter methylation of multiple genes (P = .01). CONCLUSIONS: The association between T-Ag expression and promoter methylation in CRC suggests that this viral oncogene may induce methylator phenotype and that JCV may be involved in CRC through multiple mechanisms of genetic and epigenetic instability.     

Improvement of diagnostic accuracy and screening conditions with liquid-based cytology

The aim of this population-based study was to compare the histological follow-up diagnoses of cervicocytological neoplasia (dysplasia, carcinoma in situ and carcinoma) in conventional Papanicolaou (CP) smear and ThinPrep PapTest samples (TP).All cytological samples from the County of Funen, Denmark, in the periods 2000 (n = 34,832) and 2002 (n = 29,995) were included in the study. In 2000 and 2002, the specimens were CP and TP, respectively. The detection rate of > or = mild dysplasia was 0.8% in CP and 1.4% in TP, showing a 75% increase in TP when compared with CP (p < 0.001). Histological follow-up of > or = moderate dysplasia revealed a neoplastic lesion in 77.1% and 87.9% in CP and TP, respectively (P < 0.001).The present study indicates that the diagnostic accuracy of cervical cytology is improved with liquid-based cytology. In addition, we focus on the optimized cellular material that shows the diagnostic details very clearly to the microscopist and leads to radically improved screening conditions.     

LEEP刀宫颈环形电切术治疗宫颈上皮内瘤变临床观察

目的探讨LEEP刀宫颈环形电切术治疗宫颈上皮内瘤变（CIN）的临床疗效、并发症等。方法通过对2006年1月-2008年12月行阴道镜下活检病理诊断为CIN的256例患者行LEEP刀宫颈环形电切术者随访并行回顾分析。结果阴道镜下活检病理诊断与最后诊断的符合率为88.67%（227/256）,252例患者术后1年细胞学检查均转阴性（100%）,其中245例人乳头状瘤病毒（HPV）DNA阳性患者中,238例术后1年HPVD-NA转为阴性（97.14%）。结论 LEEP刀宫颈环形电切术具有手术操作简单、并发症少、疗效高等特点,是一种理想的CIN诊治方法。     

LEEP治疗宫颈上皮内瘤变临床疗效及其预后影响因素

目的探讨LEEP对宫颈上皮内瘤变（C1N）的治疗效果及患者预后的影响因素。方法LEEP治疗267例CIN患者,分析临床效果,并根据治疗结局将患者分为治愈组与复发组,使用SPSS13．0统计软件进行统计学分析,研究影响其预后的相关因素。结果267例CIN患者均顺利完成手术,随访2年治愈率为92．1％（246／267）,复发率为7．9％（21／267）;治愈组与复发组患者在年龄、是否吸烟、术前HPV感染、术前病理结果、手术切缘状态、病变是否累及腺体以及病变范围等方面比较差异具有统计学意义（P〈0．01）,而在初次性生活年龄方面比较差异无统计学意义（P〉0．05）。结论LEEP对于CIN的治疗,具有创伤小、用时少、术后恢复快、治愈率高等特点,患者年龄、吸烟、术前HPV感染、术前病理结果、手术切缘状态、病变是否累及腺体以及病变范围为影响预后的相关因素,应给予足够重视,制定可行的手术方案,以期彻底治愈。