HPV基因的进化分析

目的:HPV有许多类型,其大致可分为高危型和低危型,高危型HPV感染是导致宫颈癌发生的首要原因,在HPV基因组中,E6基因是促进宫颈细胞癌变的关键基因,本文主要研究HPV中的E6基因在各种不同型别的HPV中的进化关系,并对E2基因碱基替换率进行分析,探讨高危型HPV与低危型HPV的区别.方法:本文对不同类型HPV E6氨基酸序列构建系统发生树,探讨识别高危型HPV可能的一致序列,对E6基因其中一处能导致恶性程度增加的突变进行分析.并对HPV16与其位于同一颗树的HPV35和HPV31计算相对碱基替换率.结果:高危型HPV均源自同一株病毒株的进化.各种HPV型别中,高危型HPV E6蛋白对应于HPV16E6蛋白的第83位氨基酸为缬氨酸更为保守,HPV中除E2以外的其他基因的非同义替换率均小于同义替换率.结论:HPV E6蛋白对应于HPV16E6蛋白的第83位氨基酸为缬氨酸能更好地实现HPV E6蛋白的致癌作用.HPV基因中除E2以外的基因在进化过程中都较为保守,是HPV增殖生长的关键基因,而E2部分区域非同义替换率大于同义替换率,说明E2这部分区域的突变能够更好的促进HPV的增殖和生长.     

扩散张量成像对颈椎病脊髓早期损伤的研究

目的:探讨磁共振(MR)扩散张量成像(DTI)作为定量分析方法,对脊髓型颈椎病(CSM)脊髓早期损伤诊断的应用价值.方法:选择45例经临床及影像诊断为脊髓型颈椎病患者,颈椎常规MRI检查显示脊髓内无异常信号,使用单次激发自旋回波平面(SE-EPI)序列,进行DTI扫描.测量压迫部位脊髓的ADC值及FA值作为病例组,选择病变上或下方两个节段以上未受压正常脊髓作为正常对照组,测量其ADC值及FA值.分析病例组与对照组间ADC及FA值差别,计算ADC值及FA值诊断脊髓损伤的敏感性.结果:所有脊髓型颈椎病患者经DTI检查均可得到ADC图及FA图,经图像后处理,脊髓显示清晰,图像无变形及伪影.3例脊髓型颈椎病患者ADC值降低,42例脊髓型颈椎病患者ADC值增高,平均ADC值为(1.388± 0.149)x 10-3 mm2/s.44名脊髓型颈椎病患者FA值降低,1名脊髓型颈椎病患者FA值增高,平均FA值为0.476±0.085,受压处脊髓平均ADC值升高,平均FA值下降,与正常值比较差别有统计学意义.ADC值诊断的敏感性为93.33％,FA值诊断的敏感性为97.78％.结论:DTI与常规MR比较,能早期而准确地诊断脊髓型颈椎病脊髓早期损伤.     

鬼臼毒素涂膜剂治疗子宫颈HPV 感染的疗效观察

目的:了解鬼臼毒素涂膜剂治疗子宫颈HPV感染的疗效。方法:126例临床诊断证明为子宫颈HPV感染的患者,随机分为两组,试验组63例采用0.5%鬼臼毒素涂膜剂治疗,对照组63例单独采用0.5%鬼臼毒素酊治疗。结果:①痊愈率:试验组与对照组的痊愈率分别为90.5%、73.00%,两组比较具有统计学学差异(P<0.05);②复发率:试验组与对照组第12、24周的复发率分别为9.52%、28.57%和9.52%、36.51%,两个时段的两组比较均具有统计学差异(P<0.05);③不良反应:经随访观察,两组发生了红斑、水肿、灼热、疼痛、瘙痒、溃疡、渗出等不良反应,其中两组比较试验组红斑、水肿、溃疡、渗出的发生率低于对照组,具有统计学差异(P<0.05)。结论:鬼臼毒素涂膜剂能有效治疗子宫颈HPV感染,并控制复发,副作用小,比采用鬼臼毒素酊治疗的效果好。     

宫颈癌患者外周血CD4+CD25+high调节性T细胞的表达及意义

目的:探讨宫颈癌患者外周血中CD4~ CD25~( high)调节性T(regulator T cells,Tr)的表达及意义。方法:采用流式细胞术检测52例宫颈癌患者,35例健康女性外周血中CD4~ CD25~( high)Tr、细胞毒性T细胞(cytotoxic T lymphocytes,CTL)和NK细胞,采用ELISA检测血清中-干扰素(interferon,IFN-)的表达水平。结果:宫颈癌患者外周血CD4~ CD25~( high)Tr占CD4~ T淋巴细胞的百分比为(7.18±2.32)%,高于健康女性组(P<0.05);宫颈癌患者外周血CD4~ CD25~( high)Tr水平与CTL、NK细胞及IFN-水平呈负相关。结论:宫颈癌患者外周血中具免疫抑制活性的CD4~ CD25~( high)Tr水平较高,参与宫颈癌患者的肿瘤免疫抑制。     

Proteomic analysis of cervical cancer cells treated with suberonylanilide hydroxamic acid

Suberonylanilide hydroxamic acid (SAHA) is an orally administered histone deacetylase inhibitor (HDACI) that has shown significant antitumour activity in a variety of tumour cells. To identify proteins involved in its antitumour activity, we utilized a proteomic approach to reveal protein expression changes in the human cervical cancer cell line HeLa following SAHA treatment. Protein expression profiles were analysed by 2-dimensional polyacrylamide gel electrophoresis (2-DE) and protein identification was performed on a MALDI-Q-TOF MS/MS instrument. As a result, a total of nine differentially expressed proteins were visualized by 2-DE and Coomassie brilliant blue (CBB) staining. Further, all the changed proteins were positively identified via mass spectrometry (MS)/MS analysis. Of these, PGAM1 was significantly downregulated in HeLa cells after treatment with SAHA. Moreover, PGAM1 has been proven to be downregulated in another cervical cancer cell line (CaSki) by western blot analysis. Together, using proteomic tools, we identified several differentially expressed proteins that underwent SAHA-induced apoptosis. These changed proteins may provide some clues to a better understanding of the molecular mechanisms underlying SAHA-induced apoptosis in cervical cancer.     

European guidelines for quality assurance in cervical cancer screening: recommendations for cytology laboratories.

The quality of a cervical cytology laboratory depends on adequate handling and staining of the samples, screening and interpretation of the slides and reporting of the results. These guidelines give an overview of procedures recommended in Europe to manage the balance between best patient care possible, laboratory quality assurance and cost effectiveness and will be published as a chapter 4 in the European Guidelines for Quality Assurance in Cervical Cancer Screening. The laboratory guidelines include protocols for personnel and organisation, material requirements, handling and analysing cervical samples, recording of results, quality management and communication. The section on quality management is comprehensive and includes protocols for all aspects of internal and external quality assurance. The guidelines are extensively referenced and as far as possible the recommendations are evidence-based.     

宫颈癌组织miR-200b-5p、miR-424-5p表达与临床病理特征、PI3K/AKT/mTOR信号通路和预后的关系研究

摘要 目的：探讨宫颈癌组织微小核糖核酸（miRNA）-200b-5p、miR-424-5p表达与临床病理特征、磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白（PI3K/AKT/mTOR）信号通路和预后的关系。方法：选取2016年7月～2019年6月西安市中心医院收治的123例宫颈癌患者,采用定量聚合酶链式反应（qPCR）检测癌组织与癌旁组织中miR-200b-5p、miR-424-5p、PI3K信使RNA（mRNA）、AKT mRNA、mTOR mRNA表达。分析miR-200b-5p、miR-424-5p表达与PI3K mRNA、AKT mRNA、mTOR mRNA表达的相关性及与临床病理特征的关系。采用K-M法绘制不同miR-200b-5p、miR-424-5p表达宫颈癌患者生存曲线。结果：与癌旁组织比较,宫颈癌组织中miR-200b-5p、miR-424-5p表达降低,PI3K mRNA、AKT mRNA、mTOR mRNA表达升高（P＜0.05）。Pearson相关性分析显示,宫颈癌组织中miR-200b-5p、miR-424-5p表达与PI3K mRNA、AKT mRNA、mTOR mRNA表达均呈负相关,PI3K mRNA、AKT mRNA、mTOR mRNA表达呈两两相关（P＜0.05）。miR-200b-5p、miR-424-5p表达与宫颈癌分化程度、国际妇产科联盟（FIGO）分期和淋巴结转移有关（P＜0.05）。随访3年,123例宫颈癌患者累积生存率为62.60%（77/123）。K-M生存曲线分析显示,miR-200b-5p、miR-424-5p高表达组累积生存率分别高于miR-200b-5p、miR-424-5p低表达组（P＜0.05）。结论：宫颈癌组织中miR-200b-5p、miR-424-5p低表达,与分化程度、FIGO分期、淋巴结转移、PI3K/AKT/mTOR信号通路和预后有关。     

The impact of health insurance affiliation and socioeconomic status on cervical cancer survival in Bucaramanga,Colombia

Cervical cancer is still an important cause of death in countries like Colombia. We aimed to determine whether socioeconomic status of residential address (SES) and type of health insurance affiliation (HIA) might be associated with cervical cancer survival among women in Bucaramanga, Colombia. All patients residing in the Bucaramanga Metropolitan Area diagnosed with invasive cervical cancer (ICD-0–3 codes C53.X) between 2008 and 2016 (n = 725) were identified through the population-based cancer registry, with 700 women having follow-up data for >5 years (date of study closure: Dec 31, 2021), yielding an overall 5-year survival estimate (95 % CI) of 56.4 % (52.7 – 60.0 %). KM estimates of 5-year overall survival were obtained to assess differences in cervical cancer survival by SES and HIA. Multivariable Cox-proportional hazards modeling was also conducted, including interaction effects between SES and HIA. Five-year overall survival was lower when comparing low vs. high SES (41.9 % vs 57.9 %, p < 0.0001) and subsidized vs. contributive HIA (45.1 % vs 63.0 %, p < 0.0001). Multivariable Cox modeling showed increased hazard ratios (HR) of death for low vs. high SES (HR = 1.78; 95 % CI = 1.18–2.70) and subsidized vs. contributive HIA (HR = 1.44; 95 % CI = 1.13–1.83). The greatest disparity in HR was among women of low SES affiliated to subsidized HIA (vs. contributive HIA and high SES) (HR=2.53; 95 % CI = 1.62–3.97). Despite Colombia’s universal healthcare system, important disparities in cervical cancer survival by health insurance affiliation and socioeconomic status remain.     

Seroreactivity to HPV-16 proteins in women with early cervical neoplasia

Summary Although serological reactivity to human papillomavirus type 16 (HPV-16) proteins has been demonstrated in patients with invasive cervical carcinoma, the degree of seroreactivity to these proteins in women with preinvasive disease and its relationship to the HPV type associated with the disease are unclear. We obtained sera from 27 women undergoing cone biopsy for cervical precursor lesions and 22 controls and analyzed seroreactivity by Western blot to fusion proteins containing portions of the HPV-16 E4, L1 and L2 open-reading frames (ORFs). Positives were analyzed by scanning densitometry and intensity values for each case plotted relative to controls. Cervical biopsy specimens from patients were analyzed for HPV-16 nucleic acids by DNA · DNA in situ hybridization. Mean intensity values for seroreactivity to the pATH-E4 protein approached significance (P = 0.058) and a significantly higher proportion of cases vs controls registered values over 4.0 for pATH-E4 (26% vs 4.5%;P = 0.04) and pATH-L2 (48% vs 18%;P = 0.03) proteins. A significantly higher mean intensity value for E4 was observed for cases containing HPV-16 DNA vs HPV-16 negative cases or controls. Thus, seroreactivity to HPV-16-derived proteins may be more common in women with preinvasive cervical disease, and for some protein targets (E4) may indicate a relatively type-specific response.Supported in part by grants from the National Cancer Institute [CA 47676 (C.P.C.)], American Cancer Society [MV-395 (C.P.C.)] and an institutional support grant (J.K.R.). Dr. Crum is a recipient of a Physician Scientist Award from the National Institute of Allergy and Infectious Disease (AI00628)     

PCR检测HPV－18DNA

采用聚合酶链式反应（ＰＣＲ）技术,对ＨＰＶ－１８序列中引物ＨＰ＿１、ＨＰ＿２之间的片段（Ｆ）进行扩增,通过两组阴、阳性对照实验证明扩增片段的特异性。用不同Ｍｇ浓度的缓冲系统进行ＰＣＲ反应发现,缓冲系统中Ｍｇ浓度高低是影响ＨＰＶ－１８／ＨＰ＿１、ＨＰ＿２特异扩增的重要因素,高浓度Ｍｇ导致扩增特异性降低。对１７例宫颈癌组织ＤＮＡ进行ＰＣＲ检测,有９例检出Ｆ片段,其检出率是５３％,为ＨＰＶ－１８与宫颈癌的相关性提供证据。